| Objective: Bladder cancer(Bladder Cancer,BLCA)refers to a malignant tumor that originates from the epithelial tissue of the bladder wall.According to statistics on a global scale in 2020,bladder cancer patients rank tenth among new cancers.Bladder cancer is divided into non-Muscle Invasive Bladder Cancer(NMIBC)and Muscle Invasive Bladder Cancer(MIBC).Compared with the former,the latter has a poor prognosis,with a lower survival rate after distant metastasis,and the 5-year survival rate is only 8.1%.The highly aggressive and highly metastatic phenotype of muscular invasive bladder cancer is the main cause of death in patients.At present,most patients with non-muscular invasive bladder cancer undergo transurethral resection of bladder tumor(TURBT)after diagnosis,followed by intravesical chemotherapy or BCG biological treatment.Among patients with muscular invasive bladder cancer,about half of the patients underwent TURBT,and a small number of patients underwent cystectomy,followed by chemotherapy and radiotherapy.However,for muscle-infiltrating bladder cancer,the results of surgery are not satisfactory,and there are still cases of metastasis and recurrence in patients after surgery.Therefore,in order to have a better treatment and prognosis for patients with bladder cancer,it is urgent to find new therapeutic targets.The JAK/STAT signal pathway is a signal transduction pathway stimulated by cytokines.The pathway is mainly composed of three components,including tyrosine kinase-related receptors,tyrosine kinase JAK and nuclear transcription factor STAT.The JAK / STAT signaling pathway can be activated by various cytokines and growth factors(such as interleukins,interferons,and EGF family members),which bind to their respective transmembrane receptors,including the tyrosine kinase JAK,the latter It is activated after ligand-induced conformational changes in the receptor.These activated tyrosine kinases,JAK,are phosphorylated and then bind to the nuclear transcription factor STAT receptor.After being phosphorylated by JAK,STAT is activated and forms a dimer into the nucleus,recognizing and binding specific promoter sequences to activate its target genes.Transcription.The JAK/STAT signaling pathway plays an important role in cell functions such as cell proliferation,stem cell maintenance and differentiation,and immune-inflammatory response.It has been reported in the literature that the JAK/STAT signaling pathway is involved in the regulation of the proliferation,invasion and metastasis of tumor cells by a variety of genes,and the JAK-STAT signaling pathway mediates the activation of T lymphocytes,thereby reducing the efficacy of anti-PD-1 immunotherapy.In bladder cancer,JAK /STAT signal pathway is an important regulatory mechanism to promote the proliferation,migration and invasion of bladder cancer,but there is no research report on the regulatory role of VGLL1 in bladder cancer.VGLL1(Vestigial-like1)is a transcriptional co-activator Vestigial(Vg)in Drosophila.The VGLLs family consists of VGLL1-4.They contain the TOUDU domain,which mediates the interaction with TEA domain transcription factors(TEADs).Some scholars have found that the structure and function of the VGLL1-TEADs and YAP-TEADs complexes are similar,and the functions of the two are basically the same.VGLL1 is highly expressed in gastric cancer,breast cancer,and ovarian cancer,and can promote the proliferation of pancreatic cancer and gastric cancer.The role of.However,VGLL4 competes with YAP to bind TEADs,and plays a role of tumor suppressor in lung cancer and bladder cancer.It has been reported in the literature that VGLL1 drives the transcription of human papillomavirus early genes by combining with TEAD1.VGLL1 can be used as a targeted placental antigen co-expressed by pancreas and basal-like breast cancer.VGLL1 is regulated by PI3K/AKT pathway.The progress and poor prognosis of gastric cancer,but the function of VGLL1 in bladder cancer has not been reported.Therefore,it is particularly important to explore the biological function and related mechanism of VGLL1 in bladder cancer.This study clarified VGLL1 as a poor prognostic factor of bladder cancer through bioinformatics and immunohistochemistry,and verified that VGLL1 enhances the proliferation,invasion and metastasis of bladder cancer cells,and further explored the regulatory mechanism of JAK/STAT3 signaling pathway on VGLL1 and enriched VGLL1 As the mechanism of oncogenes in the progression of bladder cancer,it provides a new therapeutic target for the clinical treatment of bladder cancer.Method:1.Bioinformatics analysis of the expression of VGLL1 in pan-carcinoma;the difference of expression of VGLL1 in bladder cancer tissue and normal tissue;the effect of VGLL1 on the clinical staging of bladder cancer2.Tissue chip immunohistochemical analysis of the expression difference of VGLL1 in different grades of bladder cancer and its influence on the prognosis3.CCK-8 method to detect the effect of VGLL1 on the proliferation of bladder cancer cells4.Cell scratch test to detect the effect of VGLL1 on the migration ability of bladder cancer cells5.Transwell cell invasion test detects the effect of VGLL1 on the invasion ability of bladder cancer cells6.Western Blot to detect the effect of VGLL1 on the markers of epithelial-mesenchymal transition7.GSEA gene enrichment analysis,looking for the signal pathway JAK/STAT related to VGLL18.Protein interaction network analysis,screening out the key molecule STAT3 that interacts with VGLL19.Immunofluorescence staining(IF)to detect the localization of VGLL1 and STAT3 in bladder cancer cells10.Co-immunoprecipitation(Co-IP)to detect the interaction of VGLL1 and STAT3 in bladder cancer cells11.qPCR detection of the effect of knockdown or overexpression of VGLL1 on the phosphorylation level of STAT312.Western Blot detects the effect of knockdown or overexpression of VGLL1 on the phosphorylation level of STAT313.Immunofluorescence staining(IF)to detect the effect of knockdown or overexpression of VGLL1 on the localization of STAT3 in bladder cancer cells14.Bioinformatics analysis and screening of VGLL1-STAT3 related invasion and metastasis downstream target genes MMP7,MMP915.qPCR detection of the effect of VGLL1 on the expression level of MMP7 and MMP916.Western Blot detects the effect of VGLL1 on the expression level of MMP7 and MMP9Result:1.VGLL1 has the highest expression level in pan-cancer of bladder cancer2.VGLL1 is highly expressed in bladder cancer,which is related to the stage of the disease and affects the poor prognosis3.VGLL1 promotes the proliferation,metastasis and invasion of bladder cancer cells4.GESA enrichment analysis,VGLL1 is closely related to JAK/STAT signaling pathway5.VGLL1 and STAT3 can co-bind in the nucleus of bladder cancer cells6.VGLL1 can recruit STAT3 phosphorylation into the nucleus7.VGLL1 is involved in the transcriptional regulation of downstream target genes MMP7 and MMP9 by STAT3Conclusion:1.VGLL1 is highly expressed in bladder cancer and affects poor prognosis2.VGLL1 regulates epithelial-mesenchymal transition and affects the invasion and metastasis ability of bladder cancer cells3.VGLL1 and STAT3 combine with each other and recruit STAT3 into the nucleus to participate in downstream target gene regulation4.VGLL1 is involved in the transcriptional regulation of MMP7 and MMP9 by STAT3... |
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