miR-21介导Wnt信号通路诱导胰腺癌细胞凋亡的机制研究

Mechanism of miR-21 mediating Wnt signaling pathway inducing apoptosis of pancreatic cancer cells

目的选择miR-21介导Wnt信号通路诱导胰腺癌细胞凋亡的机制研究。方法选择人胰腺腺癌细胞株ASPC-1,分为As组(正常ASPC-1细胞)、Ay组(ASPC-1细胞转染+miR-21-inhibitions)、Nc组(ASPC-1细胞转染+miR-21-阴性对照)。RT-PCR法检测Wnt1、β-catenin水平变化,Westernblot法检测Wnt1、β-catenin蛋白,Transwell法、MTT法检测ASPC-1细胞的迁移、活力变化,流式细胞仪检测ASPC-1细胞凋亡率。结果三组miR-21、Wnt1、β-catenin水平、Wnt1蛋白、β-catenin蛋白、ASPC-1细胞凋亡率比较,差异均有统计学意义(F分别=55.47、14.30、123.60、3.54、5.85、30.27,P均<0.05)。与As组、Nc组比较,Ay组miR-21、Wnt1、β-catenin水平、Wnt1蛋白、ASPC-1细胞凋亡率有明显下降,β-catenin蛋白明显上升(P均<0.05),ASPC-1细胞迁移能力明显下降。培养24 h、48 h、72 h时,三组间ASPC-1细胞活力比较,差异均有统计学意义(F分别=9.62、24.47、126.10,P均<0.05)。两两比较,培养24 h、48 h、72 h时,Ay组细胞活力均低于As组、Nc组(P均<0.05)。结论miR-21低表达能明显抑制胰腺癌细胞增殖,其作用机制可能通过抑制Wnt/β-catenin信号通路活性来实现。

Objective To study the mechanism of miR-21 mediated Wnt signaling to induce apoptosis of pancreatic cancer cells.Methods Human pancreatic adenocarcinoma cell line ASPC-1 was selected and divided into As group(normal ASPC-1 cells),Ay group(ASPC-1 cell transfection+Mir-21-inhibitions),and Nc group(ASPC-1 cell trans-fection+Mir-21-negative control).The levels of Wnt1 andβ-catenin were detected by RT-PCR,the proteins of Wnt1 andβ-catenin were detected by Westernblot,the migration and viability of ASPC-1 cells were detected by Transwell and MTT,and the apoptosis rate of ASPC-1 cells was detected by flow cytometry.Results The miR-21,Wnt1,β-catenin levels,Wnt1 protein,β-catenin protein,the apoptosis rate of ASPC-1 cells among three groups were significantly different(F=55.47,14.30,123.60,3.54,5.85,30.27,P<0.05).Compared with As group and Nc group,the miR-21,Wnt1,β-catenin levels,Wnt1 protein,the apoptosis rate of ASPC-1 cells in Ay group were decreased significantly(P<0.05)as well as the migration ability of ASPC-1 cells,while theβ-catenin protein increased obviously.At 24h,48h and 72h,the viability of ASPC-1 cells among the three groups was significantly different(F=9.62,24.47,126.10,P<0.05).The cell vi-ability of Ay group was lower than that of As group and Nc group at 24h,48 h and 72h(P<0.05).Conclusion The low expression of miR-21 can significantly inhibit the proliferation of pancreatic cancer cells,which may be achieved by inhibiting the activity of Wnt/β-catenin signaling pathway.