摘要
目的 探讨miR-145-5p靶向磷脂酶Cε1(PLCE1)对胰腺癌细胞增殖、分化和凋亡的影响。方法 qRT-PCR和Westernblot检测4种胰腺癌细胞系和正常胰腺细胞中miR-145-5p和PLCE1的表达水平。构建过表达miR-145-5p的胰腺癌细胞株,MTT法检测细胞增殖活力,流式细胞术检测细胞凋亡,Westernblot检测增殖相关蛋白、凋亡相关蛋白以及分化相关蛋白的表达。采用双荧光素酶报告基因法和Westernblot验证miR-145-5p和PLCE1的靶向关系。结果 与正常胰腺细胞相比,胰腺癌细胞中miR-145-5p表达明显降低,PLCE1的表达明显升高。过表达miR-145-5p显著抑制PANC-1细胞增殖,促进其分化和凋亡。PLCE1是miR-145-5p的靶基因,miR-145-5p可负性调控PLCE1的表达。过表达PLCE1可逆转miR-145-5p对PANC-1细胞的增殖抑制、分化和凋亡促进作用。结论 miR-145-5p通过靶向下调PLCE1抑制胰腺癌细胞的增殖,促进其分化和凋亡。
Objective To investigate the effect of miR-145-5p targeting PLCE1 on proliferation, differentiation and apoptosis of pancreatic cancer cells. Methods qRT-PCR and Western blot were used to detect the expression of miR-145-5p and PLCE1 in pancreatic cancer cell lines and normal pancreatic cells hTERT-HPNE. The pancreatic cancer cell line with the over-expression of miR-145-5p was constructed,the cell proliferation viability was detected by MTT assay,the cell apoptosis was detected by flow cytometry, and the expression of proliferation-related proteins, apoptosis-related proteins and differentiation-related proteins were detected by Western blot. The targeting relationship between miR-145-5p and miR-145-5p was verified by dual luciferase reporter gene assay and Western blot. Results Compared with normal pancreatic cancer cells,the expression of miR-145-5p in pancreatic cancer cells was significantly decreased,and the expression of PLCE1 was significantly increased. The over-expression of miR-145-5p significantly inhibited the proliferation,promoted the differentiation and apoptosis of PANC-1 cells. PLCE1 is a target gene of miR-145- 5p,and miR-145-5p negatively regulates the expression of PLCE1. The over-expression of PLCE1 reversed the Inhibition of proliferation,the promotion of differentiation and apoptosis of PANC-1 cells by miR-145-5p. Conclusion miR-145-5p inhibits the proliferation of pancreatic cancer cells and promotes its differentiation and apoptosis by down-regulating PLCE1.
作者
胡松
周文杰
徐天天
张文滔
HU Song;ZHOU Wen-jie;XU Tian-tian;ZHANG Wen-tao(Department of Surgery,the Third Reo-ple’s Hospital ofJingzhou,Jingzhou 543001)
出处
《现代消化及介入诊疗》
2019年第10期1123-1128,1132,共7页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
湖北省科学技术厅项目(EK2013D160056001525
EK2014D160088001057)
