摘要
目的观察miR-424-5P对结肠癌细胞增殖的影响,探讨Src-YAP信号通路在其中的作用。方法以结肠癌HT-29细胞作为研究对象,转染法外源性上调miR-424-5P水平,设置空白组(Blank组)、空载体组(NC组)和过表达组(miR-424-5P-OV组);RTFQ-PCR检测miR-424-5P水平;MTT法检测细胞活力;平板克隆实验检测细胞克隆形成能力;免疫荧光法检测YAP核质分布情况;Western blot法检测Src、YAP、LATS1及各自磷酸化表达水平。结果相较于NC组,miR-424-5P-OV组miR-424-5P RNA水平明显上调(P<0.01)。miR-424-5P过表达明显降低HT-29细胞活力和克隆形成能力(P<0.01)。Blank组和NC组细胞YAP以核表达为主,细胞质分布较少,而miR-424-5P-OV组细胞YAP可见明显的细胞质和细胞核分布。miR-424-5P过表达可显著上调p-YAP和p-LATS1蛋白表达水平(P<0.01),下调p-Src蛋白表达水平(P<0.01)。结论miR-424-5P过表达可抑制结肠癌HT-29细胞增殖,该作用可能与Src-YAP通路信号减弱有关。
Objective To observe the effect of miR-424-5P on Src-YAP signaling and the proliferation of colon cancer cells.Methods Colon cancer HT-29 cells were used.Exogenous up-regulation of miR-424-5P was achieved in HT-29 cells by transfection and a blank group(Blank group),an empty vector group(NC group)and an overexpression group(miR-424-5P-OV group)were prepared.RTFQ-PCR was used to detect the level of miR-424-5P.MTT assays were used to detect cell viability.The nuclear distribution of YAP was detected by immunofluorescence.Western blot was used to detect the levels of Src,YAP,LATS1 and their phosphorylation.Results Compared with the NC group,the miR-424-5P RNA level in the miR-424-5P-OV group was significantly increased(P<0.01).The overexpression of miR-424-5P significantly reduced the viability and colony-forming ability of HT-29 cells(P<0.01).In the blank and NC groups,YAP was mainly localized in the nucleus,with little cytoplasmic distribution,while in the miR-424-5P-OV group,YAP showed obvious cytoplasmic and nuclear distribution.Overexpression of miR-424-5P significantly up-regulated p-YAP and p-LATS1 protein levels(P<0.01)and down-regulated p-Src protein levels(P<0.01).Conclusions Overexpression of miR-424-5P can inhibit the proliferation of colon cancer HT-29 cells,which may be related to weakened Src-YAP signaling.
作者
刘友强
胡冀陶
李保坤
席金川
王贵英
LIU Youqiang;HU Jitao;LI Baokun;XI Jinchuan;WANG Guiying(Second Department of General Surgery,the Fourth Hospital of Hebei Medcial University,Shijiazhuang 050011;China.2.the Third Hospital of Hebei Medcial University,Shijiazhuang 050051)
出处
《中国比较医学杂志》
CAS
北大核心
2020年第9期43-48,共6页
Chinese Journal of Comparative Medicine
基金
2017年河北省医学科学研究重点课题计划(20170165)。
