普鲁卡因联合肌成束蛋白1对膀胱癌细胞的影响及机制研究

Effect of procaine combined with FSCN1 on bladder cancer cells and the underlying mechanism

目的探讨普鲁卡因联合肌成束蛋白1(FSCN1)对膀胱癌细胞增殖、凋亡和细胞周期的影响及作用机制。方法将阴性对照干扰小RNA(siRNA)和FSCN1 siRNA分别转染至人膀胱癌T24细胞,分别作为NC组和si-FSCN1 组,将仅加入LipofectamineTM 2000 的T24 细胞作为对照组。分别向NC 组和si-FSCN1 组T24 细胞中加入4 mmol/L 的普鲁卡因处理48 h,分别作为普鲁卡因组和普鲁卡因+si-FSCN1 组。噻唑蓝(MTT)法检测细胞增殖能力,流式细胞仪检测细胞周期分布情况和细胞凋亡率,蛋白质印迹法(Western blot)检测FSCN1、信号转导及转录激活因子3(STAT3)、磷酸化STAT3(p-STAT3)、细胞周期蛋白D1(cyclin D1)、Bcl-2 相关X蛋白(BAX)蛋白的相对表达量。结果普鲁卡因组、si-FSCN1组和普鲁卡因+si-FSCN1组细胞G0/G1期细胞比例、细胞凋亡率和BAX蛋白的相对表达量均高于对照组细胞,G2/M 期及S 期细胞比例、A值、p-STAT3 和cyclin D1 蛋白的相对表达量均低于对照组细胞,差异均有统计学意义(P﹤0.05);且普鲁卡因+si-FSCN1 组细胞G0/G1期细胞比例、细胞凋亡率和BAX蛋白的相对表达量均高于普鲁卡因组和si-FSCN1 组细胞,G2/M 期及S 期细胞比例、A值、p-STAT3 和cyclin D1蛋白的相对表达量均低于普鲁卡因组和si-FSCN1组细胞,差异均有统计学意义(P﹤0.05)。结论普鲁卡因或FSCN1 均可抑制膀胱癌细胞增殖,诱导细胞凋亡,将肿瘤细胞阻滞于G0/G1期,且可能通过STAT3 信号通路发挥协同作用。

Objective To investigate the effects of procaine combined with fascin 1 (FSCN1) on the proliferation, apoptosis and cell cycle of bladder cancer cells. Method Negative control interfering small RNA (siRNA) and FSCN1 siRNA were transfected into human bladder cancer T24 cells, as NC group and si-FSCN1 group, respectively, additionally, T24 cells added with Lipofectamine. 2000 were used as control group. Procaine at 4 mmol/L was added to the NC group and the si-FSCN1 group to treat for 48 h, as the procaine group and the procaine+si-FSCN1 group, respectively, the control group to which procaine was not added were not treated specially. The proliferation of cells was detected by methylthiazolyldiphenyl tetrazolium (MTT) assay. Cell cycle distribution and apoptosis rate were detected by flow cytometry. Western blot was used to detect the relative expression levels of FSCN1, signal transduction and activator of transcription 3 (STAT3), phospho-STAT3 (p-STAT3), cyclin D1, and Bcl-2-associated X protein (BAX). Result The proportion of G0/G1 phase cells, apoptosis rate, and relative expression of BAX proteins in procaine group, si-FSCN1 and procaine+ si-FSCN1 group were significantly higher than that in the control group, but the proportion of G2/M phase and S phaes cells, A value, relative expression of p-STAT3 and cyclin D1 proteins were lower compared to control group, the differences were of statistical significance (P<0.05). Besides, in procaine+si-FSCN1 group, the proportion of G0/G1 phase cells, cell apoptosis rate and relative expression level of BAX proteins were higher than those in procaine group and si-FSCN1 group, while the proportion of G2/M phase cells, S phase cells, A value, relative expressions of p-STAT3 and cyclin D1 proteins were lower compared to procaine group and the si-FSCN1 group, indicating statistically significant differences (P<0.05). Conclusion Procaine or FSCN1 can inhibit the proliferation of bladder cancer cells, induce apoptosis, arrest tumor cells in G0/G1 phase, and may play a synergistic role through STAT3 signaling pathway.