PRL-3表达载体构建及其对结直肠癌细胞生长的影响

摘要

Objective:To construct and identify expression vector for upregulating the level in colorectal cancer cell SW620,and to investigate its effects on the growth of SW620 cells.Methods:The total RNA of Hela cells was ex-tracted and reversed to cDNA.Then,PRL-3 gene was amplified using PCR and cloned into the pcDNA3.1 vector. The resulting plasmid was used to transfect SW620 cells.Next,qPCR and Western blot were performed to examine the expression level of PRL-3 in SW620 cells.CCK-8,colony formation assays and FCM were used to study the effects of PRL-3 on the growth of SW620 cells.Results:PRL-3 plasmid was efficiently expressed in SW620 cells,and CCK-8,colony formation assays and FCM indicated that overexpression of PRL-3 can promote the proliferation of SW620 cells.Conclusion:Expression vector of PRL-3 was successfully constructed,and upregulation of PRL -3 promoted the growth of SW620 cells.%目的:构建PRL-3过表达载体,研究其对结直肠癌细胞系SW620生长的影响.方法:提取Hela细胞总RNA并反转为cDNA,以cDNA为模板通过PCR反应扩增PRL-3基因,将扩增产物克隆至表达载体pcDNA3.1,通过转染让SW620细胞过表达PRL-3,空载体组和空白组作为阴性对照.qPCR、Western blot实验用来检测PRL-3在SW620细胞的表达水平,CCK-8实验、平板克隆形成实验用来检测SW620细胞的增殖状况,流式细胞术用来检测SW620细胞的周期分布.结果:成功构建了PRL-3过表达载体.与空载体组和空白组相比,PRL-3过表达载体转染结直肠癌细胞系SW620后,qPCR检测PRL-3表达水平上调数倍.CCK-8实验表明过表达PRL-3能够促进SW620细胞增殖.平板克隆形成实验表明上调PRL-3能够增强SW620细胞增殖能力.流式细胞术结果表明过表达PRL-3能够加快SW620细胞周期进程.结论:PRL-3能够促进结直肠癌细胞系SW620生长,为结直肠癌早期诊断和靶向治疗提供实验依据.