Cancer Translational Medicine

Original Research | Open Access

Vol.8 (2022) | Issue-3 | Page No: 104-115


Increased Expression of IL-17A, IL-6, STAT3, TGF-β, and VEGF: Potential Biomarkers in Bladder Cancer?

Zishen Xiao1, Chengxia Bai1, Teng Zhao1, Jiayu Lin1, Lijuan Yang1, Jian Liu2, Zhenjiang Wang1, Ying Sun3,4, Yanbo Liu1*


1. School of Basic Medical Sciences, Beihua University, Jilin City, Jilin, China

2. People's hospital in Yushu town, Yushu, Jilin, China

3. Department of Immunology, School of Basic Medical Science, Capital Medical University, Beijing, China

4. Division of Asthma, Allergy & Lung Biology, King’s College London, Guy’s Hospital, London, United Kingdom

*Corresponding Author

Address for correspondence: Prof. Yanbo Liu, School of Basic Medical Sciences, Beihua University, No.3999 East Binjiang Road, Fengman District, Jilin City 132013, Jilin, China. E-mail address:

Important Dates  

Date of Submission:   15-Aug-2022

Date of Acceptance:   13-Sep-2022

Date of Publication:   29-Sep-2022


Aim: To explore the association of IL-17A, IL-6, STAT3, VEGF, TGF-β, and MMP-9 with the severity of bladder cancer, and analyze the pathogenesis of bladder cancer.

Methods: Immunohistochemistry (IHC) analyzed the expression and location of IL-17A, IL-6, STAT3, and VEGF in bladder cancer specimens with different degrees of malignancy (n = 80), cystitis (n = 23), and normal adjacent tissues (n = 4). ELISA measured the serum concentrations of MMP-9, TGF-β, VEGF, and IFN-γ in bladder cancer patients (n = 34) and controls (n = 50).

Results: IL-17A, IL-6, STAT3, and VEGF were significantly increased in bladder cancer tissues compared to both cystitis (P = 0.001) and normal adjacent tissues (P = 0.001), and were positively associated with the degree of malignancy. Serum concentrations of TGF-β and VEGF were significantly higher in bladder cancer patients compared with controls (P = 0.002 and P = 0.0001, respectively), while concentrations of MMP-9 and IFN-γ were not significantly different among groups. MMP-9, VEGF, and TGF-β serum concentration increased with the severity of malignancy, and the difference between low-grade and high-grade malignancy was significant (P = 0.001, P = 0.011, and P = 0.030, respectively), IFN-γ serum secretion was lower in high-grade compared to low-grade malignancy (P = 0.014).

Conclusion: Elevated expression of IL-17A, IL-6, and STAT3 in tissues and elevated serum concentration of TGF-β, and VEGF might be considered potential biomarkers of bladder carcinoma progression.


Trial registration: ISRCTN, ISRCTN2012BH006. Registered 10 January 2012


Keywords: Bladder cancer, interleukin-17, interleukin-6, interferon-γ, vascular endothelial growth factor


Bladder cancer is the second most common urological malignancy, accounting for 5% of all cancer-related deaths in the USA.[1] It is estimated that there were approximately 76,960 new cases of bladder cancer and 16,390 deaths in the year 2016.[2] The incidence in males is about fourfold higher than that in females, while the incidence is double in white men compared to their black counterparts. Cystitis is considered a high risk for bladder cancer.[3] It has been shown that hyperplasia and chronic cystitis are easy to develop bladder cancer.[4] Although many factors including long-term smoking, rubber, leather, dye, and aluminum polluted work environment may be associated with the pathogenesis, some cytokines such as interleukin-17 (IL-17) family, interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), transforming growth factor-β (TGF-β) and signal transducer and activator of transcription 3 (STAT3) are considerably involved in the occurrence and development of cystitis and bladder cancer.[2],[5],[6] An increase in inflammatory mediators or proinflammatory cytokines has been shown to lead to tumorigenesis, invasion, angiogenesis, and prognosis. The collecting evidence shows that proinflammatory cytokines, such as IL-17A family, tumor necrosis factor-alpha (TNF-α), IL-6, VEGF, etc. play important roles in bladder cancer. Cytokine IL-17A can activate IL-6 in inflammatory or cancerous disease, while IL-6, a major activator of STAT3 signaling pathways, is the most important cytokine influencing the inflammatory response in humans.[7],[8] IL-17A, IL-6, and STAT3 signals have been implicated in the regulation of tumor growth and metastatic spread, which are also associated with prognosis in different cancers.[9],[10],[11] Furthermore, it has been reported that the increased concentrations of matrix metalloproteinase-9 (MMP-9), TGF-β, and VEGF in serum were associated with metastasis and poor prognosis of different kinds of malignant tumors.[12],[13],[14] In addition, interferon-γ (IFN-γ) may play a role in the control of tumor growth and metastasis, positively by enhancing tumor immunogenicity.[15] Although the collecting data suggest that IL-17A, IL-6, STAT3, TGF-β, and VEGF may be critical cytokines in various malignancies, the expression, and association of these molecules with bladder cancer remain unclear. In the present study, we focused on the expression of IL-17A, IL-6, VEGF, and STAT3 in tissues of the bladder, the serum concentrations of MMP-9, TGF-β, VEGF, and IFN-γ, and their association with the severity of bladder cancer.


Patients and specimens

The present study was approved by the institution of Hospital Ethics Committees of Urinary System Diseases Prevention and Treatment Research Centre in Affiliated Hospital of Beihua University, Jilin City, Jilin Province, People’s Republic of China (approval reference: 2012BH006). Written consent was obtained from the participants. The tissue specimens were collected from patients with bladder cancer (n = 80, including 4 adjacent normal tissues) and patients with cystitis (n = 23) between January 2012 and December 2014. All the tissue samples were identified and diagnosed by two clinical pathologists with a double-blind method. The clinical characteristics of the subjects involved in this study are summarized in Table 1, which is the same as our previously published study.[5]

Table 1.
Table 1. Clinical characteristics of the subjects from cystitis and bladder cancer

Serum samples were collected from 34 preoperative patients with bladder cancer and 50 age-matched healthy volunteers. None of the bladder cancer patients received radiotherapy or chemotherapy before surgery, while the healthy volunteers had no immunological or infectious diseases of any kind. Informed consent was obtained from each participant for the use of blood samples in the present study [Table 2].

Table 2.
Table 2. Clinical characteristics of the subjects from bladder cancer and healthy volunteers


Immunohistochemistry was employed to measure immunoreactivity for IL-17A, IL-6, STAT3, and VEGF following a protocol as previously described.[5] Briefly, a household pressure cooker was used to retrieve antigens at high temperature and high pressure with sodium citrate solution (0.01 mmol/L, pH = 6.0). Endogenous peroxidase activity was inhibited using freshly prepared hydrogen peroxide in methanol (0.3%) at room temperature for 30 min. Slides were washed with PBS and then blocked with 2.5% horse serum blocking buffer (Vector Laboratories, Cat# S-2012) for 20 minutes and then with dilution buffer containing 5% goat serum for a further 30 min at room temperature. Proposed immunoreactivity was measured using antibodies against IL-17A (Novus Biological, USA, NBPI-42746, 1:100), IL-6 (Novus Biological, NBPI-42746, 1:800), VEGF (Beijing Biosynthesis Biotechnology CO, LTD, Beijing, China, bs-1141R, 1:500) and STAT3 (Beijing Biosynthesis Biotechnology CO, LTD, bs-1141R, 1:500). The information of antibodies used in the present study are summarized in Table 3. DAB kit (diaminobenzidine, ZhongShan Golden Bridge Biological Company, Beijing, China) was used to detect positive signals (brown). The stained slides were observed and scanned using an Olympus microscope at 10× magnification. These images were exported as TIFF files and uploaded into Image Pro Plus 6.0 software (Media Cybernetics, Maryland, USA) for further analysis. The brown positive signals were quantified as the percentages of IL-17A, IL-6, STAT3, and VEGF immunoreactive staining of the total hematoxylin counterstaining area of the entire sections as described previously.[5]

Table 3. Antibodies or reagent kits used in this research
Table 3. Antibodies or reagent kits used in this research

Enzyme-linked immunosorbent assay (ELISA)

ELISA kits were purchased from eBioscience, Bender MedSystems GmbH, Vienna, Austria, for measuring concentrations of MMP-9 (LOT, 98474017, sensitivity: 50 pg/mL), TGF-β (LOT, 98482009, sensitivity: 8.6 pg/mL), VEGF (LOT, 98478015, sensitivity: <5 pg/mL) and IFN-γ (LOT, 98476080, sensitivity: < 4 pg/mL) according to the manufacturer’s instructions [Table 3].

Statistical analysis

Data obtained from immunohistochemistry were analyzed with a commercially available statistical package (Minitab for Windows, Minitab Release 9.2; Minitab, Inc, State College, PA). Differences between groups were analyzed using the Kruskal-Wallis test followed by the Mann-Whitney U test. Data from ELISA assays were analyzed using the Student t-test and one-way analysis of variance (ANOVA). Data are presented as the mean ± SEM. A P value of less than 0.05 was considered statistically significant.


Expression and location of IL-17A and IL-6 in bladder tissues

The immunohistochemical staining analysis showed that global IL-17A immunoreactivity was significantly higher in tissue sections from bladder cancer and cystitis compared with normal tissues (P = 0.001 and P = 0.001, respectively) Figure 1A and B]. In addition, immunoreactivity for IL-17A was significantly higher in tissue sections of bladder cancer than that of cystitis (P = 0.001). Immunoreactivity for IL-17A was mainly located in mononuclear cells, transitional epithelial cells, malignant cells, and vascular endothelial cells in bladder cancer Figure 1A].

IL-6, as the more important downstream cytokine of IL-17A, is the primary pro-inflammatory cytokine in humans and is produced primarily by T-lymphocytes and macrophages. We previously showed that there were more infiltrating macrophages in bladder cancer tissues.[5] So we measure the expression and location of IL-6 in tissues of patients with bladder cancer compared with that of control subjects. Immunohistochemistry revealed that immunoreactivity for IL-6 was significantly elevated in sections of tissues of bladder cancer compared with that of cystitis and normal tissues (P = 0.001 and P = 0.001, respectively) Figure 1C and D], while IL-6 immunoreactivity in sections of tissues of cystitis was also significantly higher in cystitis than that of normal tissues (P = 0.024). IL-6 immunoreactivity was predominantly located in monocytes, transitional epithelial cells, malignant cells as well as vascular endothelial cells [Figure 1C].

Figure 1.
Figure 1. Expression and location of IL-17A and IL-6 in tissue sections of cystitis and bladder cancer. A: Representative photomicrographs of tissue sections showing immunoreactivity to IL-17A in subjects with cystitis (n = 23), bladder cancer (n = 80) and adjacent normal tissues (n = 4) (original magnification × 10 and 20). B: Quantitative analysis of immunoreactive area of IL-17A in tissue sections (% of whole sections). C: Representative photomicrographs of immunoreactivity for IL-6 in tissue sections of subjects with cystitis, bladder cancer and adjacent normal tissues (original magnification × 10 and 20). D: Quantitative analysis of immunoreactive area of IL-6 in tissue sections (% of whole sections). Data are expressed as the mean ± SEM. Arrows show examples of positively stained cells. The scale bar means 100 μm.


Expression and location of VEGF and STAT3 in bladder tissues

Previous research results have confirmed that VEGF acted as an important regulator in cell proliferation, and metastasis in many types of malignant tumors,[16],[17] which is upregulated by IL-17A and IL-6. In this case, we measured immunoreactivity for VEGF in tissues of bladder cancer and compared them with the controls. Immunohistochemical staining analysis showed that the immunoreactivity for VEGF was significantly higher in tissue sections from bladder cancer compared with the controls (P = 0.001) [Figure 2A and B]. Although the expression of VEGF in cystitis was elevated, it did not achieve statistical significance when it was compared with that of normal tissues (P = 0.446). Immunoreactivity for VEGF was mainly located in vascular endothelial cells of tumor parenchyma compared to the stroma of tumors [Figure 2A].

STAT3 is linked to inflammation-related oncogenesis and is constitutively activated in various cancers. Persistent activation of STAT3 is involved in promoting tumor cell proliferation, survival, tumor invasion, angiogenesis, and immunosuppression, while IL-6 is considered to up-regulate and activate STAT3.[18],[19] Again, our immunohistochemical staining analysis showed that STAT3 immunoreactivity was significantly higher in bladder cancer tissues compared with the controls (P = 0.001) [Figure 2C and D], while there was no significant difference between cystitis and normal tissues (P = 0.4172). STAT3 was mainly located in malignant cells, vascular endothelial cells, and transitional epithelial cells [Figure 2C].

Figure 2.
Figure 2. Expression and location of VEGF and STAT3 in tissue sections of cystitis and bladder cancer. A: Representative photomicrographs of tissue sections showing immunoreactivity to VEGF in subjects with cystitis (n = 23), bladder cancer (n = 80) and adjacent normal tissues (n = 4) (original magnification × 10 and 20). B: Quantitative analysis of immunoreactive area of VEGF in tissue sections (% of whole sections). C: Representative photomicrographs of immunoreactivity for STAT3 in tissue sections of subjects with cystitis, bladder cancer and adjacent normal tissues (original magnification × 10 and 20). D: Quantitative analysis of immunoreactive area of STAT3 in tissue sections (% of whole sections). Data are expressed as the mean ± SEM. Arrows show examples of positively stained cells. The scale bar means 100 μm.


Association between the expression of IL-17A, IL-6, VEGF, and STAT3 and severity of bladder cancer

Further investigation showed that tissue sections from the median and late stages (High-grade) had considerably higher levels of immunoreactivities for IL-17A, IL-6, STAT3, and VEGF than those from the early stages (Low-grade) (P = 0.012, P = 0.014, P = 0.034, P = 0.024, respectively) [Figure 3A-D].

IL-17A, IL-6, STAT3, and VEGF expression levels were correlated using the Pearson correlation method. The obvious positive correlation was observed between IL-17A and IL-6 (r = 0.5931), IL-17A and STAT3 (r = 0.6374), IL-6 and STAT3 (r = 0.3963). Furthermore, the IL-6 expression in bladder cancer was associated with that of VEGF (r = 0.3968) which indicated that the IL-17A-IL-6-VEGF signal axis might participate in the occurrence and progression of bladder cancer.

Figure 3.
Figure 3. The relationships between immunoreactivities for IL-17A, IL-6, STAT3 and VEGF and malignancy in bladder cancer. A: The association of immunoreactivity for IL-17A with bladder cancer malignancy. B: The association of immunoreactivity for IL-6 with bladder cancer malignancy. C: The association of immunoreactivity for STAT3 with bladder cancer malignancy. D: The association of immunoreactivity for VEGF with bladder cancer malignancy. Data are expressed as the mean ± SEM.


Serum MMP-9

Total concentrations of MMP-9 were not significantly elevated in patients with bladder cancer compared with that of the control subjects (166.11 ± 28.66 vs 162.85 ± 4.25 pg/mL; P = 0.636) [Figure 4A]. It is interesting, however, to note that the mean concentration of serum MMP-9 of patients at the early disease stages (Low-grade) was significantly lower than that of patients at the median and late stages (High-grade) (152.15 ± 5.57 and 184.38 ± 5.57 pg/mL respectively, P=0.001) [Figure 4B]. There was no significant difference in concentrations of MMP-9 between male and female patients (P > 0.05).

Serum IFN-γ

Similarly, although there was no significant difference in the mean concentration of serum IFN-γ between patients with bladder cancer and control subjects (7.01 ± 0.61 vs 7.45 ± 0.62 pg/mL respectively; P = 0.606) [Figure 4C], the mean concentration of serum IFN-γ of patients at the median and late stages (High-grade) was significantly lower than that of early stages (Low-grade) (5.65 ± 0.87 vs 8.18 ± 0.12 pg/mL, P = 0.014) [Figure 4D]. Again, serum concentrations of IFN-γ were not significantly different between male and female patients (P > 0.05).

Figure 4.
Figure 4. Concentrations of serum MMP-9, IFN-γ in bladder cancer patients and control subjects. Concentrations of serum MMP-9 and IFN-γ were measured retrospectively in patients with bladder cancer (n = 34) and controls (n = 50) using the commercial ELISA kits. Data are expressed the mean ± SEM.


Serum VEGF

Unlike MMP9 and IFN-γ, the mean concentration of serum VEGF of bladder cancer patients was significantly higher than that of control subjects (479.55 ± 40.38 vs 233.15 ± 30.93 pg/mL, P = 0.0001) [Figure 5A]. In addition, the mean concentration of serum VEGF was significantly higher in the median and late stages (High-grade) than that of early stages (Low-grade) (629.92 ± 83.75 vs 364.56 ± 55.35 pg/mL respectively, P = 0.011) [Figure 5B]. Again, there was no significant difference in concentrations of VEGF between male and female patients (P > 0.05).

Serum TGF-β

Similar to VEGF, the mean concentration of serum TGF-β was significantly elevated in patients with bladder cancer than that of control subjects (1308.28 ± 89.65 vs 910.10 ± 65.22 pg/mL, P = 0.002) [Figure 5C]. In addition, serum TGF-β was significantly lower in the early stages of bladder cancer (Low-grade) than that of median and late stages (High-grade) (1088.70 ± 87.85 vs 1595.41 ± 227.38 pg/mL, P = 0.030) [Figure 5D]. Again, there was no significant difference in serum concentrations of TGF-β between male and female patients (P > 0.05).

Figure 5.
Figure 5. Concentrations of serum VEGF and TGF-β in bladder cancer patients and control subjects. Concentrations of serum VEGF and TGF-β were measured retrospectively in patients with bladder cancer (n = 34) and controls (n = 50) using the commercial ELISA kits. Data are expressed the mean ± SEM.


It is known that urothelial cell carcinomas constitute approximately 95% of all bladder cancer cases,[20] while early diagnosis and timely treatment are very pivotal for increasing the 5-year survival rate.[21],[22] Thus, any potential biomarkers for predicting the prognosis and malignancy of the disease are worth to be investigated. Our previous studies proved that elevated expression of IL-17A was observed in tissues derived from bladder cancer, prostate cancer, and rectal carcinoma suggesting that this cytokine may promote the occurrence and development of malignant diseases.[5],[23],[24] However, the association between IL-17A and its downstream cytokines such as IL-6, VEGF, and STAT3, has not been systematically investigated. In the present study, immunoreactivities for IL-17, IL-6, VEGF, and STAT3 were significantly higher in tissues of bladder cancer than that of cystitis and controls and were also associated with malignancy of the disease. A common hallmark of cystitis and bladder cancer due to altered microbiota is the infiltration of pervasive inflammatory cells, which may generate an effective and specific inflammatory response by synthesizing and eliminating more cytokines and inflammatory markers including IL-17A and IL-6. These abnormally elevated cytokines and humoral factors may induce the production of more inflammatory cytokines leading to severe inflammatory response and tissue damage, forming a typical positive feedback process. Moreover, persistent activation of STAT3 can maintain constitutive VEGF activity, thus providing evidence for the relation between oncology signaling pathways within the inflammatory microenvironment.[25] On the other hand, these cytokines might also promote tumorigenesis, tumor growth, and even metastasis through inhibiting tumor suppressor genes or activating oncogenes such as STAT3, TGF-β, and MMP-9. Our data revealed that there existed an association between the expression of these molecules and the severity of malignancy, which further suggests the importance of these molecules in the pathogenesis of the disease.

It has been shown that IL-17A can increase the growth and proliferation of cervical cancer cells via IL-6 or has the potential to act as a prognostic biomarker for the progression of colorectal cancer.[26],[27] Increased IL-6 may act as the main activator of the JAK/STAT3 signaling pathway contributing to tumor proliferation, angiogenesis, and vascular modeling through VEGF and STAT3 activation.[28],[29],[30] Our immune analysis showed that an elevated IL-6 expression and serum VEGF concentration are associated with malignancy of bladder cancer. A correlation has been shown between an elevated expression of IL-6 and decreased response to treatment, shorter survival time, and increased disease failure rates.[31],[32],[33],[34],[35] In addition, our research results showed that, in bladder cancer, the immunoreactivity to IL-17A was positively correlated with IL-6, further IL-17A was also associated with STAT3. Interestingly, IL-6 expression levels correlated positively with STAT3 and VEGF. All of the results indicated that IL-17A regulated the synthesis and secretion of IL-6, and IL-6 further regulated the expression of STAT3 and VEGF. Hence, the IL-17A-IL-6-VEGF axis played a vital role. Since cancer is a chronic inflammatory disease, it promotes the infiltration of inflammatory cells in the cancerous tissue. Those inflammatory cells such as lymphocytes, monocytes, phagocytes, neutrophils, etc. could release many kinds of cytokines, such as IL-17A, IL-1, TNF-α, etc. In turn, those inflammatory cytokines further activated their receptor, directly or indirectly activating the downstream corresponding elements (oncogene or the genes which promote cancer development and even metastasis).

Apart from VEGF, increased concentrations of serum TGF-β and their association with malignancy of bladder cancer were also observed in the present study. In this case, elevated TGF-β and VEGF in the late stage of cancer may imply cancer deterioration. It has been known that malignant neoplasms and T cells can produce a large amount of TGF-β at the late stages of most solid cancers.[30] Therefore, targeting cytokines IL-17A, IL-6, VEGF and TGF-β might provide some clinical benefits for patients suffering from bladder cancer. On the other hand, levels of expression of these cytokines might also be used as biomarkers for predicting tumor progression and recurrence.

It is also well known that MMP-9, VEGF, and TGF-β, as downstream products of STAT3 or IL-6, can be secreted into serum or local environment to participate in invasion or metastasis of malignancy or to prevent cancer progression.[11],[36],[37],[38] MMP-9 is a potent proteolytic enzyme and plays a key role in the degradation of basal membranes and the extracellular matrix, through cleaving type IV collagen and gelatin, which are the main structural components of the basal membrane. The proteolytic activity of MMP‑9 is not only to induce invasion and metastasis but also to generate matrix-bound growth factors and other signaling molecules responsible for growth signaling, angiogenesis, and inflammatory response.[39],[40],[41] Again, ELISA results showed that serum concentration of MMP-9 is associated with malignant grade of bladder cancer. This is possibly relevant to elevated expression of IL-6 because the IL-6 silencing vector decreases expressions of VEGF, MMP-9, and STAT3.[42],[43]

IFN-γ, a kind of cytokine, can promote not only immunomodulation but also anticancer activity. IFN-γ binds to its receptor and subsequently activates its downstream signaling transcriptional pathways which are principally involved in its biological activities. Regarding IFN-γ-dependent immunosurveillance, IFN-γ can directly suppress tumorigenesis and/or can modulate the immunological status of cancer cells and infected cells.[44] The collected findings show that endogenous IFN-γ not only controls tumorigenesis and progression but also shapes the immunogenicity of tumors and promotes the growth of tumor cells with immune evolutionary properties.[45] Whether IFN-γ is anti-tumorigenic or pro-tumorigenic remains controversial.[46],[47],[48] Our research results showed that IFN-γ levels were a litter lower in serum samples of bladder cancer patients than that of healthy individuals, but its levels were significantly lower in high-grade bladder cancer than that in low-grade, which indicated that IFN-γ has an antitumor role in bladder cancer. The mechanism may involve IFN-γ secreted by natural killer (NK) cells and cytotoxic T lymphocytes, which recruit various cells of innate and adaptive immunity to tumor sites and promote their activation function. It is also very well known that IFN-γ enhances antigenicity of tumor cells via up-regulation of the major histocompatibility complex (MHC) class Ia membrane expression. IFN-γ could stimulate the expression of tumor antigen-presenting MHC molecules to increase the immunogenicity of tumor cells and makes them more susceptible to immune recognition and destruction.[49] IFN-γ also displays direct anticancer activity via inhibition of cell proliferation, e.g., by upregulation of p21 and p27 molecules to arrest the cell cycle, or through the mediation of apoptotic cell death.[50],[51] Moreover, by targeting non-transformed cells present in the tumor microenvironment, IFN-γ displays its indirect anti-tumor actions, acting as an antiangiogenic factor to inhibit tumor angiogenesis and/or to promote the destruction of established tumor-associated blood vessels.

Although we have done a series of comparative immunoreactivities for IL-17A, IL-6, VEGF, and STAT3 in bladder cancer and “normal” tissues and serum concentrations of MMP-9, IFN-γ, VEGF, and TGF-β in patients with bladder cancer and control subjects, the present study has obvious limitations. Firstly, the specimens used were obtained through endoscopic biopsy or resection, which might limit our measurements. Secondly, the specimens and serum samples were not completely marched. Finally, we did not collect the follow-up data, so the expression and concentrations of these molecules might not well reflect the prognosis of the diseases. Certainly, subsequent studies will be carried out; particularly more samples from subjects with low-grade, early-stage urologic diseases.

In summary, our data suggest that elevated expression of IL-17A, IL-6, STAT3, VEGF, TGF-β, and MMP-9 may participate in the pathogenesis of bladder cancer. The Association of these molecules with malignant grades suggests that these molecules might be considered as biomarkers, either for diagnosis and prognosis, or for therapeutic purpose, or both. IFN-γ would be seen as a kind of antitumor cytokine in bladder cancer deterioration.


Elevated expression of IL-17A, IL-6, and STAT3 in tissues and of TGF-β, and VEGF in serum might be considered as potential biomarkers for clinical stages of bladder carcinoma progression.



All the authors thank Prof. Hongwen Gao’s team in the pathology department, at the Second Hospital of Jilin University for their help in recruiting patients, collecting specimens, documenting clinical information and pathological diagnosis, and we also appreciate Jing Jiang from the affiliated hospital of Beihua University for her contribution to collect serum and tissue specimens.



This work was supported by the Project of Jilin Provincial Department of science and technology to Yanbo Liu (No.20210402015GH, No. YDZJ202201ZYTS194), Jilin Provincial Department of Education to Lijuan Yang (No.JJKH20220068KJ); Graduate Innovation Program of Beihua University to Zishen Xiao (No.2021020); Graduate Innovation Program of Beihua University to Chengxia Bai (No.2021055).



The authors declare that they have no competing interests.



The study was approved by the Hospital Ethics Committees of Urinary System Diseases Prevention and Treatment Research Centre, the Affiliated Hospital of Beihua University (approval number: 2012BH006). Written informed consent was obtained from each subject who participated in the study. We confirmed that we have read the editorial policy and included relative ethics questions in the appropriate place in the present manuscript.



Yanbo Liu designed the experiments and drafted the manuscript. Zishen Xiao and Teng Zhao performed immunohistochemistral staining of IL-17A, IL-6, STAT3 and VEGF cytokines. Jiayu Lin performed the Elisa assays. Chengxia Bai and Lijuan Yang carried out the statistical analysis. Zhenjiang Wang provided the experimental place and participated in coordination and supervision. Jian Liu provided some experimental samples. Ying Sun conceived the study and modified the manuscript. All authors have read and approved the final manuscript.



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Hatel Rana Moonat, Gangxiong Huang, Pooja Dhupkar, Keri Schadler, Nancy Gordon,
Eugenie Kleinerman

Efficacy and Safety of Paclitaxel‑based Therapy and Nonpaclitaxel‑based Therapy in Advanced Gastric Cancer

Tongwei Wu, Xiao Yang, Min An, Wenqin Luo, Danxian Cai, Xiaolong Qi

Motion Estimation of the Liver Based on Deformable Image Registration: A Comparison Between Four‑Dimensional‑Computed Tomography and Four‑Dimensional-Magnetic Resonance Imaging

Xiao Liang1, Fang‑Fang Yin1,2, Yilin Liu1, Brian Czito2, Manisha Palta2, Mustafa Bashir3, Jing Cai1,2

A Feasibility Study of Applying Thermal Imaging to Assist Quality Assurance of High‑Dose Rate Brachytherapy

Xiaofeng Zhu1, Yu Lei1, Dandan Zheng1, Sicong Li1, Vivek Verma1, Mutian Zhang1, Qinghui Zhang1, Xiaoli Tang2, Jun Lian2, Sha X. Chang2, Haijun Song3, Sumin Zhou1, Charles A. Enke1

Role of Exosome microRNA in Breast Cancer

Wang Qu, Ma Fei, Binghe Xu

Recent Progress in Technological Improvement and Biomedical Applications of the Clustered Regularly Interspaced Short Palindromic Repeats/Cas System

Yanlan Li1,2*, Zheng Hu1*, Yufang Yin3, Rongzhang He1, Jian Hu1, Weihao Luo1, Jia Li1, Gebo Wen2, Li Xiao1, Kai Li1, Duanfang Liao4, Di-Xian Luo1,5

The Significance of Nuclear Factor‑Kappa B Signaling Pathway in Glioma: A Review

Xiaoshan Xu1, Hongwei Yang2, Xin Wang2, Yanyang Tu1

Markerless Four‑Dimensional‑Cone Beam Computed Tomography Projection‑Phase Sorting Using Prior Knowledge and Patient Motion Modeling: A Feasibility Study

Lei Zhang1,2, Yawei Zhang2, You Zhang1,2,3, Wendy B. Harris1,2, Fang‑Fang Yin1,2,4, Jing Cai1,4,5, Lei Ren1,2

The Producing Capabilities of Interferon‑g and Interleukin‑10 of Spleen Cells in Primary and Metastasized Oral Squamous Cell Carcinoma Cells-implanted Mice

Yasuka Azuma1,2, Masako Mizuno‑Kamiya3, Eiji Takayama1, Harumi Kawaki1, Toshihiro Inagaki4, Eiichi Chihara2, Yasunori Muramatsu5, Nobuo Kondoh1

“Eating” Cancer Cells by Blocking CD47 Signaling: Cancer Therapy by Targeting the Innate Immune Checkpoint

Yi‑Rong Xiang, Li Liu

Glycosylation is Involved in Malignant Properties of Cancer Cells

Kazunori Hamamura1, Koichi Furukawa2

Biomarkers in Molecular Epidemiology Study of Oral Squamous Cell Carcinoma in the Era of Precision Medicine

Qing‑Hao Zhu1*, Qing‑Chao Shang1*, Zhi‑Hao Hu1*, Yuan Liu2, Bo Li1, Bo Wang1, An‑Hui Wang1

I‑Kappa‑B Kinase‑epsilon Activates Nuclear Factor‑kappa B and STAT5B and Supports Glioblastoma Growth but Amlexanox Shows Little Therapeutic Potential in These Tumors

Nadège Dubois1, Sharon Berendsen2, Aurélie Henry1,2, Minh Nguyen1, Vincent Bours1,
Pierre Alain Robe1,2

Suppressive Effect of Mesenchymal Stromal Cells on Interferon‑g‑Producing Capability of Spleen Cells was Specifically Enhanced through Humoral Mediator(s) from Mouse Oral Squamous Cell Carcinoma Sq‑1979 Cells In Vitro

Toshihiro Inagaki1,2, Masako Mizuno‑Kamiya3, Eiji Takayama1, Harumi Kawaki1, Eiichi Chihara4, Yasunori Muramatsu5, Shinichiro Sumitomo5, Nobuo Kondoh1

An Interplay Between MicroRNA and SOX4 in the Regulation of Epithelial–Mesenchymal Transition and Cancer Progression

Anjali Geethadevi1, Ansul Sharma2, Manish Kumar Sharma3, Deepak Parashar1

MicroRNAs Differentially Expressed in Prostate Cancer of African‑American and European‑American Men

Ernest K. Amankwah

The Role of Reactive Oxygen Species in Screening Anticancer Agents

Xiaohui Xu1, Zilong Dang2, Taoli Sun3, Shengping Zhang1, Hongyan Zhang1

Panobinostat and Its Combination with 3‑Deazaneplanocin‑A Induce Apoptosis and Inhibit In vitro Tumorigenesis and Metastasis in GOS‑3 Glioblastoma Cell Lines

Javier de la Rosa*, Alejandro Urdiciain*, Juan Jesús Aznar‑Morales, Bárbara Meléndez1,
Juan A. Rey2, Miguel A. Idoate3, Javier S. Castresana

Cancer Stem‑Like Cells Have Cisplatin Resistance and miR‑93 Regulate p21 Expression in Breast Cancer

Akiko Sasaki1, Yuko Tsunoda2, Kanji Furuya3, Hideto Oyamada1, Mayumi Tsuji1, Yuko Udaka1, Masahiro Hosonuma1, Haruna Shirako1, Nana Ichimura1, Yuji Kiuchi1

The Contribution of Hexokinase 2 in Glioma

Hui Liu1, Hongwei Yang2, Xin Wang3, Yanyang Tu1

The Mechanism of BMI1 in Regulating Cancer Stemness Maintenance, Metastasis, Chemo‑ and Radiation Resistance

Xiaoshan Xu, Zhen Wang, Nan Liu, Pengxing Zhang, Hui Liu, Jing Qi, Yanyang Tu

A Multisource Adaptive Magnetic Resonance Image Fusion Technique for Versatile Contrast Magnetic Resonance Imaging

Lei Zhang1,2, Fang‑Fang Yin1,2,3, Brittany Moore1,2, Silu Han1,2, Jing Cai1,2,4

Senescence and Cancer

Sulin Zeng1,2, Wen H. Shen2, Li Liu1

The “Wild”‑type Gastrointestinal Stromal Tumors: Heterogeneity on Molecule Characteristics and Clinical Features

Yanhua Mou1, Quan Wang1, Bin Li1,2

Retreatment with Cabazitaxel in a Long‑Surviving Patient with Castration‑Resistant Prostate Cancer and Visceral Metastasis

Raquel Luque Caro, Carmen Sánchez Toro, Lucia Ochoa Vallejo

Therapy‑Induced Histopathological Changes in Breast Cancers: The Changing Role of Pathology in Breast Cancer Diagnosis and Treatment

Shazima Sheereen1, Flora D. Lobo1, Waseemoddin Patel2, Shamama Sheereen3,
Abhishek Singh Nayyar4, Mubeen Khan5

Glioma Research in the Era of Medical Big Data

Feiyifan Wang1, Christopher J. Pirozzi2, Xuejun Li1

Transarterial Embolization for Hepatocellular Adenomas: Case Report and Literature Review

Jian‑Hong Zhong1,2, Kang Chen1, Bhavesh K. Ahir3, Qi Huang4, Ye Wu4, Cheng‑Cheng Liao1, Rong‑Rong Jia1, Bang‑De Xiang1,2, Le‑Qun Li1,2

Nicotinamide Phosphoribosyltransferase: Biology, Role in Cancer, and Novel Drug Target

Antonio Lucena‑Cacace1,2,3, Amancio Carnero1,2

Enhanced Anticancer Effect by Combination of Proteoglucan and Vitamin K3 on Bladder Cancer Cells

Michael Zhang, Kelvin Zheng, Muhammad Choudhury, John Phillips, Sensuke Konno

Molecular Insights Turning Game for Management of Ependymoma: A Review of Literature

Ajay Sasidharan, Rahul Krishnatry

IDH Gene Mutation in Glioma

Leping Liu1, Xuejun Li1,2

Challenges and Advances in the Management of Pediatric Intracranial Germ Cell Tumors: A Case Report and Literature Review

Gerard Cathal Millen1, Karen A. Manias1,2, Andrew C. Peet1,2, Jenny K. Adamski1

Assessing the Feasibility of Using Deformable Registration for Onboard Multimodality‑Based Target Localization in Radiation Therapy

Ge Ren1,2,3, Yawei Zhang1,2, Lei Ren1,2

Research Advancement in the Tumor Biomarker of Hepatocellular Carcinoma

Qing Du1, Xiaoying Ji2, Guangjing Yin3, Dengxian Wei3, Pengcheng Lin1, Yongchang Lu1,
Yugui Li3, Qiaohong Yang4, Shizhu Liu5, Jinliang Ku5, Wenbin Guan6, Yuanzhi Lu7

Novel Insights into the Role of Bacterial Gut Microbiota in Hepatocellular Carcinoma

Lei Zhang1, Guoyu Qiu2, Xiaohui Xu2, Yufeng Zhou3, Ruiming Chang4

Central Odontogenic Fibroma with Unusual Presenting Symptoms

Aanchal Tandon, Bharadwaj Bordoloi, Safia Siddiqui, Rohit Jaiswal

The Prognostic Role of Lactate in Patients Who Achieved Return of Spontaneous Circulation after Cardiac Arrest: A Systematic Review and Meta‑analysis

Dongni Ren1, Xin Wang2, Yanyang Tu1,2

Inhibitory Effect of Hyaluronidase‑4 in a Rat Spinal Cord Hemisection Model

Xipeng Wang1,2, Mitsuteru Yokoyama2, Ping Liu3

Research and Development of Anticancer Agents under the Guidance of Biomarkers

Xiaohui Xu1, Guoyu Qiu1, Lupeng Ji2, Ruiping Ma3, Zilong Dang4, Ruling Jia1, Bo Zhao1

Idiopathic Hypereosinophilic Syndrome and Disseminated Intravascular Coagulation

Mansoor C. Abdulla

Phosphorylation of BRCA1‑Associated Protein 1 as an Important Mechanism in the Evasion of Tumorigenesis: A Perspective

Guru Prasad Sharma1, Anjali Geethadevi2, Jyotsna Mishra3, G. Anupa4, Kapilesh Jadhav5,
K. S. Vikramdeo6, Deepak Parashar2

Progress in Diagnosis and Treatment of Mixed Adenoneuroendocrine Carcinoma of Biliary‑Pancreatic System

Ge Zengzheng1, Huang-Sheng Ling2, Ming-Feng Li2, Xu Xiaoyan1, Yao Kai1, Xu Tongzhen3,
Ge Zengyu4, Li Zhou5

Surface-Enhanced Raman Spectroscopy to Study the Biological Activity of Anticancer Agent

Guoyu Qiu1, Xiaohui Xu1, Lupeng Ji2, Ruiping Ma3, Zilong Dang4, Huan Yang5

Alzheimer’s Disease Susceptibility Genes in Malignant Breast Tumors

Steven Lehrer1, Peter H. Rheinstein2

OSMCC: An Online Survival Analysis Tool for Merkel Cell Carcinoma

Umair Ali Khan Saddozai1, Qiang Wang1, Xiaoxiao Sun1, Yifang Dang1, JiaJia Lv1,2, Junfang Xin1, Wan Zhu3, Yongqiang Li1, Xinying Ji1, Xiangqian Guo1

Protective Activity of Selenium against 5‑Fluorouracil‑Induced Nephrotoxicity in Rats

Elias Adikwu, Nelson Clemente Ebinyo, Beauty Tokoni Amgbare

Advances on the Components of Fibrinolytic System in Malignant Tumors

Zengzheng Ge1, Xiaoyan Xu1, Zengyu Ge2, Shaopeng Zhou3, Xiulin Li1, Kai Yao1, Lan Deng4

A Patient with Persistent Foot Swelling after Ankle Sprain: B‑Cell Lymphoblastic Lymphoma Mimicking Soft‑tissue Sarcoma

Crystal R. Montgomery‑Goecker1, Andrew A. Martin2, Charles F. Timmons3, Dinesh Rakheja3, Veena Rajaram3, Hung S. Luu3

Coenzyme Q10 and Resveratrol Abrogate Paclitaxel‑Induced Hepatotoxicity in Rats

Elias Adikwu, Nelson Clemente Ebinyo, Loritta Wasini Harris

Progress in Clinical Follow‑up Study of Dendritic Cells Combined with Cytokine‑Induced Killer for Stomach Cancer

Ling Wang1,2, Run Wan1,2, Cong Chen1,2, Ruiliang Su1,2, Yumin Li1,2

Supraclavicular Lymphadenopathy as the Initial Manifestation in Carcinoma of Cervix

Priyanka Priyaarshini1, Tapan Kumar Sahoo2

ABO Typing Error Resolution and Transfusion Support in a Case of an Acute Leukemia Patient Showing Loss of Antigen Expression

Debasish Mishra1, Gopal Krushna Ray1, Smita Mahapatra2, Pankaj Parida2

Protein Disulfide Isomerase A3: A Potential Regulatory Factor of Colon Epithelial Cells

Yang Li1, Zhenfan Huang2, Haiping Jiang3

Clinicopathological Association of p16 and its Impact on Outcome of Chemoradiation in Head‑and‑Neck Squamous Cell Cancer Patients in North‑East India

Srigopal Mohanty1, Yumkhaibam Sobita Devi2, Nithin Raj Daniel3, Dulasi Raman Ponna4,
Ph. Madhubala Devi5, Laishram Jaichand Singh2

Potential Inhibitor for 2019‑Novel Coronaviruses in Drug Development

Xiaohui Xu1, Zilong Dang2, Lei Zhang3, Lingxue Zhuang4, Wutang Jing5, Lupeng Ji6, Guoyu Qiu1

Best‑Match Blood Transfusion in Pediatric Patients with Mixed Autoantibodies

Debasish Mishra1, Dibyajyoti Sahoo1, Smita Mahapatra2, Ashutosh Panigrahi3

Characteristics and Outcome of Patients with Pheochromocytoma

Nadeema Rafiq1, Tauseef Nabi2, Sajad Ahmad Dar3, Shahnawaz Rasool4

Comparison of Histopathological Grading and Staging of Breast Cancer with p53‑Positive and Transforming Growth Factor‑Beta Receptor 2‑Negative Immunohistochemical Marker Expression Cases

Palash Kumar Mandal1, Anindya Adhikari2, Subir Biswas3, Amita Giri4, Arnab Gupta5,
Arindam Bhattacharya6

Chemical Compositions and Antiproliferative Effect of Essential Oil of Asafoetida on MCF7 Human Breast Cancer Cell Line and Female Wistar Rats

Seyyed Majid Bagheri1,2, Davood Javidmehr3, Mohammad Ghaffari1, Ehsan Ghoderti‑Shatori4

Cyclooxygenase‑2 Contributes to Mutant Epidermal Growth Factor Receptor Lung Tumorigenesis by Promoting an Immunosuppressive Environment

Mun Kyoung Kim1, Aidin Iravani2, Matthew K. Topham2,3

Potential role of CircMET as A Novel Diagnostic Biomarker of Papillary Thyroid Cancer

Yan Liu1,2,3,4#, Chen Cui1,2,3,4#, Jidong Liu1,2,3,4, Peng Lin1,2,3,4,Kai Liang1,2,3,4, Peng Su5, Xinguo Hou1,2,3,4, Chuan Wang1,2,3,4, Jinbo Liu1,2,3,4, Bo Chen6, Hong Lai1,2,3,4, Yujing Sun1,2,3,4* and Li Chen 1,2,3,4*

Cuproptosis-related Genes in Glioblastoma as Potential Therapeutic Targets

Zhiyu Xia1,2, Haotian Tian1, Lei Shu1,2, Guozhang Tang3, Zhenyu Han4, Yangchun Hu1*, Xingliang Dai1*

Cancer Diagnosis and Treatments by Porous Inorganic Nanocarriers

Jianfeng Xu1,2, Hanwen Zhang1,2, Xiaohui Song1,2, Yangong Zheng3, Qingning Li1,2,4*

Delayed (20 Years) post-surgical Esophageal Metastasis of Breast Cancer - A Case Report

Bowen Hu1#, Lingyu Du2#, Hongya Xie1, Jun Ma1, Yong Yang1*, Jie Tan2*

Subtyping of Undifferentiated Pleomorphic Sarcoma and Its Clinical Meaning

Umair Ali Khan Saddozai, Zhendong Lu, Fengling Wang, Muhammad Usman Akbar, Saadullah Khattak, Muhammad Badar, Nazeer Hussain Khan, Longxiang Xie, Yongqiang Li, Xinying Ji, Xiangqian Guo

Construction of Glioma Prognosis Model and Exploration of Related Regulatory Mechanism of Model Gene

Suxia Hu, Abdusemer Reyimu, Wubi Zhou, Xiang Wang, Ying Zheng, Xia Chen, Weiqiang Li, Jingjing Dai

ESRP2 as a Non-independent Potential Biomarker-Current Progress in Tumors

Yuting Chen, Yuzhen Rao, Zhiyu Zeng, Jiajie Luo, Chengkuan Zhao, Shuyao Zhang

Resection of Bladder Tumors at the Ureteral Orifice Using a Hook Plasma Electrode: A Case Report

Jun Li, Ziyong Wang, Qilin Wang

Structural Characterization and Bioactivity for Lycium Barbarum Polysaccharides

Jinghua Qi1,2,  Hangping Chen3,Huaqing Lin2,4,Hongyuan Chen1,2,5* and Wen Rui2,3,5,6*

The Role of IL-22 in the Prevention of Inflammatory Bowel Disease and Liver Injury

Xingli Qi1,2, Huaqing Lin2,3, Wen Rui2,3,4,5 and Hongyuan Chen1,2,3

RBM15 and YTHDF3 as Positive Prognostic Predictors in ESCC: A Bioinformatic Analysis Based on The Cancer Genome Atlas (TCGA)

Yulou Luo1, Lan Chen2, Ximing Qu3, Na Yi3, Jihua Ran4, Yan Chen3,5*

Mining and Analysis of Adverse Drug Reaction Signals Induced by Anaplastic Lymphoma Kinase-Tyrosine Kinase Inhibitors Based on the FAERS Database

Xiumin Zhang1,2#, Xinyue Lin1,3#, Siman Su1,3#, Wei He3, Yuying Huang4, Chengkuan Zhao3, Xiaoshan Chen3, Jialin Zhong3, Chong Liu3, Wang Chen3, Chengcheng Xu3, Ping Yang5, Man Zhang5, Yanli Lei5*, Shuyao Zhang1,3*

Advancements in Immunotherapy for Advanced Gastric Cancer

Min Jiang1#, Rui Zheng1#, Ling Shao1, Ning Yao2, Zhengmao Lu1*

Tumor Regression after COVID-19 Infection in Metastatic Adrenocortical Carcinoma Treated with Immune Checkpoint Blockade: A Case Report

Qiaoxin Lin1, Bin Liang1, Yangyang Li2, Ling Tian3*, Dianna Gu1*

Mining and Analysis of Adverse Events of BRAF Inhibitors Based on FDA Reporting System

Silan Peng1,2#, Danling Zheng1,3#, Yanli Lei4#, Wang Chen3, Chengkuan Zhao3, Xinyue Lin1, Xiaoshan Chen3, Wei He3, Li Li3, Qiuzhen Zhang5*, Shuyao Zhang1,3*

Malignant Phyllodes Tumor with Fever, Anemia, Hypoproteinemia: A Rare and Strange Case Report and Literature Review

Zhenghang Li1, Yuxian Wei1*

Construction of Cuproptosis-Related LncRNA Signature as a Prognostic Model Associated with Immune Microenvironment for Clear-Cell Renal Cell Carcinoma

Jiyao Yu1#, Shukai Zhang2#, Qingwen Ran3, Xuemei Li4,5,6*

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