TPX2基因沉默对膀胱癌耐药细胞株T24/DDP顺铂化疗敏感性的增强作用及其机制OA北大核心CSTPCD

Objective:To discuss the effect of gene silencing of targeting protein for Xenopus kinesin like protein 2(TPX2)on the chemosensitivity of the resistant bladder cancer cell line T24/cisplatin(DDP),and to clarify the mechanism.Methods:The DDP-resistant cell line T24/DDP was established by DDP concentration gradient induction method,and the cells were divided into T24 cell group and T24/DDP cell group.MTT method was used to detect the proliferation activities of the cells in various groups;the resistance index(RI)was calculated based on the half maximal inhibitory concentration(IC50)value;real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the expression levels of TPX2 mRNA and protein in the cells in various groups;small interfering RNA(siRNA)was used to silence the TPX2 gene expression in the T24/DDP cells.The cells were divided into blank control group,negative control siRNA(si-NC)group,TPX2 silenced(si-TPX2)group,si-NC+ DDP(2 mg·L-1 DDP)group,and si-TPX2+DDP(2 mg·L-1 DDP)group.RT-qPCR and Western blotting methods were used to detect the expression levels of TPX2 mRNA and protein in the cells in various groups;MTT method was used to detect the proliferation activity of the cells in various groups;flow cytometry was used to detect the apoptotic rates of the cells and percentages of the cells at G2/M phase in various groups;Transwell chamber assay was used to detect the numbers of migration and invasion cells;Western blotting method was used to detect the expression levels of Wnt/β-catenin signaling pathway-related proteins such as β-catenin,P-glycoprotein(P-gp),zinc finger protein transcription factor 1(Snail1),and Survivin proteins in the cells in various groups.Results:The resistant bladder cancer cell line T24/DDP was successfully established with the RI value of 8.76.Compared with T24 cell group,the expression levels of TPX2 mRNA and protein in the T24/DDP cells were significantly increased(P<0.01).Compared with blank control group and si-NC group,the expression levels of TPX2 mRNA and protein in the T24/DDP cells in si-TPX2 group were significantly decreased(P<0.01),and the IC50 value of DDP was significantly decreased(P<0.01).Compared with si-NC group,the apoptotic rate of the cells and the percentage of the cells at G2/M phase in si-TPX2 group was significantly increased(P<0.01),and the numbers of migration and invasion cells were significantly decreased(P<0.01),and the expression levels of β-catenin,P-gp,Snail1,and Survivin proteins in the T254/DDP cells were also significantly decreased(P<0.01).Compared with si-NC+DDP group,the apoptotic rate of the cells and percentage of the cells at G2/M phase in si-TPX2+DDP group were significantly increased(P<0.01),and the numbers of migration and invasion cells were significantly decreased(P<0.01),and the expression levels of β-catenin,P-gp,Snail1,and Survivin proteins in the T24/DDP cells were significantly decreased(P<0.01).Conclusion:Gene silencing of TPX2 enhances the chemosensitivity of the resistant bladder cancer cell line T24/DDP to DDP by inhibiting the Wnt/β-catenin signaling pathway.