高表达miR-331-3p下调PI3K/AKT通路抑制人肺癌A549细胞增殖  

Overexpression of miR-331-3p down-regulates PI3K/AKT pathway and inhibits proliferation of human lung cancer A549 cells

作　　者：伊敏努尔·吐尔逊 张茜 白文梅 YIMINNUER Tu-Er-Xun;ZHANG Qian;BAI Wen-Mei(Respiratory Department of Traditional Chinese Medicine Hospital of Xinjiang Autonomous Region,Urumqi 830000,China)

机构地区：[1]新疆自治区中医医院呼吸科,乌鲁木齐830000

出　　处：《中国免疫学杂志》2021年第6期689-693,共5页Chinese Journal of Immunology

摘　　要：目的:探讨高表达miR-331-3p对PI3K/AKT通路及肺癌A549细胞增殖的影响。方法:体外培养正常肺上皮BEAS-2B细胞和肺癌A549细胞,qRT-PCR检测miR-331-3p表达水平。阳离子脂质体LipofectamineTM 2000瞬时转染miR-331-3p mimics和mimics NC至A549细胞为mimics阴性对照组和miR-331-3p mimics组,细胞不做任何处理为空白对照组。48 h采用qRT-PCR法检测miR-331-3p表达情况;CCK-8法检测细胞增殖情况;平板克隆实验检测菌落形成情况;Western blot法检测增殖细胞核抗原(PCNA)、细胞核相关抗原(Ki67)、脂酰肌醇3-激酶-蛋白激酶B(PI3K)、蛋白激酶B(PKB,又称AKT)、p-AKT蛋白表达情况。瞬时转染si-PI3K和si-NC后Western blot法检测PI3K蛋白表达情况。结果:与正常肺上皮BEAS-2B细胞比较,肺癌A549细胞中miR-331-3p表达水平降低(P<0.05)。与空白组、mimic阴性对照组相比,miR-331-3p mimics组miR-331-3p表达水平升高(P<0.05);36 h、48 h、60 h、72 h OD450降低(P<0.05);细胞克隆数量、PCNA、Ki67、PI3K、p-AKT蛋白表达水平降低(P<0.05)。与空白组、siRNA NC组相比,siR-PI3K组PI3K的蛋白表达量降低,miR-331-3p表达水平升高(P<0.05)。结论:高表达miR-331-3p可能通过下调PI3K/AKT通路抑制人肺癌A549细胞增殖。Objective:To study the effect of overexpression of miR-331-3p regulates PI3K/AKT pathway and inhibits prolifer⁃ation of lung cancer A549 cells.Methods:Normal lung epithelial BEAS-2B cells and lung cancer A549 cells were cultured in vitro,qRT-PCR was used to detect the expression of miR-331-3p in cells.The cationic liposome LipofectamineTM 2000 was transfected instan⁃taneously into the miR-331-3p mimics and the miR-NC negative control group(mimics NC)of lung cancer A549 cells,without any treatment as blank group,the expression of miR-331-3p was detected by qRT-PCR 48 hours later;CCK-8 method was used to detect cell proliferation;plate cell cloning assay was used to detect colony formation;the expressions of proliferating cell nuclear antigen(PCNA),nuclear associated antigen(Ki67),lipoinositol 3-kinase-protein kinase B(PI3K),protein kinase B(PKB namely AKT),p-AKT were detected by Western blot.transfected instantaneously into the si-PI3K and si-NC,the expressions of PI3K by Wester blot.Results:Compared with normal lung epithelial BEAS-2B cells,the expression of miR-331-3p in lung cancer A549 cells was signifi⁃cant lower(P<0.05).Compared with blank group and mimics NC group,the expression level of microRNA-331-3p in mimics group in⁃creased(P<0.05),OD450 decreased at 36 h,48 h,60 h and 72 h(P<0.05),and the number of cell clones,the expression levels of PCNA,Ki67,PI3K and p-AKT proteins were decreased(P<0.05).Compared with blank group and siRNA NC group,the protein ex⁃pression of PI3K in the siR-PI3K group was decreased,miR-331-3p expression level was elevated(P<0.05).Conclusion:Effects of high expression of may inhibit the proliferation of human lung cancer A549 cells by down-regulating the PI3K/AKT pathway.

关 键 词：miR-331-3p 肺癌 脂酰肌醇3-激酶-蛋白激酶B/蛋白激酶B通路 增殖 

分 类 号：R734.2[医药卫生—肿瘤]