摘要
目的 分析miR-145在结直肠癌组织中的表达,探讨高表达miR-145对结直肠癌细胞增殖和侵袭能力的影响及其可能机制。方法 采用qRT-PCR检测miR-145在结直肠癌组织中的表达。将miR-145 mimics及其对照序列转染结肠癌HCT116细胞,采用MTT法检测各组HCT116细胞增殖能力,Transwell实验检测各组HCT116细胞侵袭能力。通过靶基因预测网站Targetscan7.2预测ZEB2是否为miR-145的靶基因,双荧光素酶报告基因实验进行验证,Western blot检测各组HCT116细胞中ZEB2蛋白表达。结果 qRT-PCR结果显示,与癌旁组织相比,结直肠组织miR-145低表达(P<0.05)。转染miR-145模拟物后,miR-145表达升高,ZEB2蛋白的表达下降(P<0.05),HCT116细胞的增殖能力和侵袭能力下降(P<0.05)。TargetScan7.2网站分析及双荧光素酶实验证实ZEB2为miR-145的靶基因。结论 miR-145在结直肠癌组织中的表达下降,高表达miR-145可以抑制结肠癌HCT116细胞增殖及侵袭能力,其机制可能与下调ZEB2的表达有关。
Objective To analyze the expression of miR-145 in colorectal cancer tissue and explore the effect of high expression of miR-145 on the proliferation and invasion of colorectal cancer cells and its possible mechanism.Methods The expression of miR-145 in colorectal cancer tissue was detected by qRT-PCR. miR-145 mimics and miRNC were transfected into colon cancer HCT116 cells. The transfection efficiency was verified by qRT-PCR. Western blot was used to detect the effect of up-regulation of miR-145 on ZEB2 expression. MTT was used to detect the proliferation of HCT116 cells. Transwell assay was used to detect the invasion of HCT116 cells. The target gene prediction website Targetscan 7.2 was used to predict whether ZEB2 was the target gene of miR-145. The dual luciferase reporter gene experiment was used to verify the prediction. Results qRT-PCR results showed than the level of miR-145 in colorectal tissues was lower than that in adjacent tissues(P<0.05). After transfection of miR-145 mimics, the expression of miR-145was increased, the expression of ZEB2 protein was decreased(P<0.05), and the proliferation and invasion ability of HCT116cells was decreased(P<0.05). TargetScan7.2 website analysis and double luciferase assay confirmed that ZEB2 was the target gene of miR-145. Conclusion The expression of miR-145 in colorectal cancer tissues was decreased, and overexpression of miR-145 could inhibit the proliferation and invasion of colon cancer HCT116 cells, and its mechanism may be related to the down-regulation of ZEB2 expression.
作者
李泽强
张成
赵金
李壮
刘艳
李井野
LI Ze-qiang;ZHANG Cheng;ZHAO Jin;LI Zhuang;LIU Yan;LI Jing-ye(General Surgery,Benxi Central Hospital,Benxi 117099;General surgery,General Hospital of Norther Theater Command,Shenyang 110016,China)
出处
《解剖科学进展》
CAS
2022年第2期200-203,共4页
Progress of Anatomical Sciences
基金
辽宁省自然科学基金面上项目(原优秀人才培育)(20170540951)。