MAP3K14下调miR-17-5p对胰腺癌细胞增殖和凋亡的影响

Effects of MAP3K14 on the proliferation and apoptosis of pancreatic cancer cells by down-regulating the expression of miR175p

目的研究MAP3K14在胰腺癌组织中的表达及对胰腺癌细胞增殖、凋亡的影响,并探讨其分子机制。方法采用RT-PCR法检测胰腺癌组织和正常胰腺组织中MAP3K14的表达。利用脂质体转染胰腺癌细胞,实验分组:Con组、pcDNA3.1组、pcDNA3.1-MAP3K14组、pcDNA3.1-MAP3K14+miR-NC组、pcDNA3.1-MAP3K14+miR-17-5p组。流式细胞术和CCK-8法分别检测细胞的凋亡、增殖。采用RT-PCR法检测转染后胰腺癌Panc-1细胞中miR-17-5p的表达。流式细胞术和CCK-8法分别检测细胞的凋亡、增殖。采用双荧光素酶报告基因实验检测MAP3K14对miR-17-5p的靶向作用。结果与人正常胰腺组织比较,胰腺癌组织中MAP3K14的表达显著降低,MAP3K14过表达可抑制胰腺癌Panc-1、MIAPaCa-2细胞的增殖,促进细胞的凋亡。双荧光素酶报告实验证实MAP3K14对miR-17-5p具有靶向作用,MAP3K14可负向调控miR-17-5p的表达。miR-17-5p过表达可显著逆转MAP3K14对Panc-1细胞增殖、凋亡的影响。结论MAP3K14可通过下调miR-17-5p抑制胰腺癌细胞的增殖、促进细胞凋亡。

Objective To investigate the expression of MAP3K14 in pancreatic cancer tissues and its effects on the proliferation and apoptosis of human pancreatic cancer cells,and to explore its molecular mechanism.Methods The expression levels of MAP3K14 in pancreatic cancer tissues and normal pancreatic tissues were detected by RT PCR.The pancreatic cancer cells were transfected by liposome.The experiment was divided into control roup,pcDNA3.1 group,pcDNA3.1 MAP3K14 group,pcDNA3.1 MAP3K14+miR NC group and pcDNA3.1 MAP3K14+miR 175p group.The apoptosis and proliferation of cells were detected by flow cytometry and CCK 8 method,respectively.The expression levels of miR175p in pancreatic cancer Panc1 cells were detected by RT PCR.The targeting effects of MAP3K14 for miR175p were detected by dual luciferase reporter gene assay.Results The expression levels of MAP3K14 in pancreatic ancer tissues were significantly decreased,as compared with those in normal pancreatic tissue.The overexpression of MAP3K14 inhibited the proliferation of pancreatic cancer Panc1 and MIAPaCa2 cells and promoted the cells apoptosis.The dual luciferase reporter assay showed that MAP3K14 had targeting effects on miR175p,and MAP3K14 can negatively regulate the expression of miR175p.Moreover the overexpression of miR175p could significantly reversed the effects of MAP3K14 on proliferation and apoptosis of Panc1 cells.Conclusion MAP3K14 can inhibit the proliferation of pancreatic cancer cells and promote cell apoptosis by down-regulating the expression of miR175p.