摘要
目的本文旨在明确miR-216a是否通过靶向调控PKCα表达而抑制胃癌细胞增殖和促进其凋亡,从而进一步揭示miR-216a的抑瘤分子机制。方法首先构建PKCα3′UTR-荧光素酶报告载体,通过荧光素酶报告检测观察miR-216a对PKCα3′UTR-荧光素酶活性的影响;将miR-216a模拟物转染胃癌细胞MGC-803,采用Western blot检测PKCα蛋白表达水平;将PKCαsiRNA转染MGC-803,通过MTS细胞增殖活性检测和Annexin V-FITC/PI凋亡实验考察PKCα下调对MGC-803增殖和凋亡的影响。结果荧光素酶报告检测显示,miR-216a能特异性地与PKCα3′UTR结合,抑制其荧光素酶活性,下调36%。过表达miR-216a的MGC-803PKCα蛋白表达水平降低。siRNA干扰PKCα表达能抑制MGC-803的增殖和侵袭能力,它能部分模拟miR-216a的抑瘤功能。结论 miR-216a通过靶向PKCαmRNA 3′UTR而抑制胃癌细胞增殖和侵袭能力。
Objective To confirm whether miR-216a suppresses cell proliferation and induces cell apoptosis by targeting PKCα, thus to reveal molecular mechanism that miR-216a functions as a tumor suppressor in gastric cancer .Methods PKCα3′untranslat-ed region（UTR）-luciferase vector was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-216a on luciferase activity .MGC-803 cells were transfected with miR-216a mimics ,and next Western blotting was performed to detect the expression of PKCαprotein .The effects of PKCαdownregulation on cell proliferation and apoptosis were observed after PKCαsiRNA were transfected into MGC-803 cells .MGC-803 cell proliferation assays were performed when cotransfected with miR-216a mimics .Results The result demonstrated miR-216a could bind to the 3′UTR of PKCαand inhibited the luciferase activi-ty ,cut the 41% .PKCαprotein expressions were significantly down-regulated when miR-216a was overexpressed in MGC-803 .siR-NA-mediated downregulation of PKCα could suppress the potentials of cell proliferation and induce apoptosis .Conclusion miR-216a suppresses cell proliferation and induces apoptosis by targeting PKCαmRNA 3′UTR in gastric cancer .
出处
《重庆医学》
CAS
CSCD
北大核心
2013年第33期4051-4052,4055,共3页
Chongqing medicine
基金
国家自然科学基金资助项目(81107526)
