摘要
目的探讨lncRNA LINC00958对结直肠癌细胞凋亡及放射敏感性的影响以及其作用机制。方法将pcDNA、pcDNA-LINC00958、si-NC、si-LINC00958、miR-NC和miR-422a质粒分别转染到SW480细胞中,并分别记为pcDNA组、pcDNA-LINC00958组、si-NC组、si-LINC00958组、miR-NC组和miR-422a组;将anti-miR-NC和anti-miR-422a质粒分别与si-LINC00958共转染到SW480细胞中,并分别记为si-LINC00958+anti-miR-NC组和si-LINC00958+anti-miR-422a组;分别将miR-NC和miR-422a分别转染到WT-LINC00958和MUT-LINC00958组细胞中,检测荧光活性;转染均用脂质体法。采用qRT-PCR检测miR-422a和LINC00958的表达;Western blot检测蛋白表达;流式细胞术检测细胞凋亡;细胞克隆形成实验检测对结直肠癌细胞放射敏感性的影响;双荧光素酶报告基因检测实验检测荧光活性。结果结直肠癌细胞中LINC00958高表达,miR-422a低表达;抑制LINC00958表达和过表达miR-422a,可促进结直肠癌细胞凋亡,并增加细胞放射敏感性。LINC00958可靶向调节miR-422a表达;抑制miR-422a,逆转了抑制LINC00958表达对结直肠癌细胞的放射增敏和细胞凋亡促进的作用。结论抑制LINC00958表达,增加结直肠癌细胞的放射敏感性,并促细胞凋亡,其机制可能与调控miR-422a有关,将为结直肠癌治疗提供新靶点和新思路。
Objective To investigate the effect of lncRNA LINC00958 on the apoptosis and radiosensitivity of colorectal cancer cells and its underlying mechanism.Methods The pcDNA,pcDNA-LINC00958,si-NC,si-LINC00958,miR-NC,and miR-422a plasmids were transfected into SW480 cells and assigned into the pcDNA group,pcDNA-LINC00958 group,si-NC group,si-LINC00958 group,miR-NC group,miR-422a group,respectively.Anti-miR-NC and anti-miR-422a plasmids were co-transfected into SW480 cells with si-LINC00958,and assigned into the si-LINC00958+anti-miR-NC group and si-LINC00958+anti-miR-422a group.miR-NC and miR-422a were transfected into the WT-LINC00958 and MUT-LINC00958 cells,respectively.The fluorescence activity was detected.Cell transfection was performed by liposome method.The expression levels of miR-422a and LINC00958 were measured by qRT-PCR.The expression levels of proteins were detected by Western Blot.Cell apoptosis was assessed by flow cytometry.The radiosensitivity of colorectal cancer cells was evaluated by cell clone formation assay.The fluorescence activity was detected by dual luciferase reporter assay.Results High expression of LINC00958 and low expression of miR-422a were observed in colorectal cancer cells.Inhibition of LINC00958 expression and overexpression of miR-422a could promote cell apoptosis and increase cell radiosensitivity of colorectal cancer cells.LINC00958 could target the regulation of miR-422a expression.Inhibition of miR-422a reversed the effect of inhibiting the expression of LINC00958 on increasing the radiosensitization and promoting cell apoptosis of colorectal cancer cells.Conclusions Inhibition of LINC00958 expression increases the radiosensitivity and promotes the apoptosis of colorectal cancer cells.The mechanism may be related to the regulation of miR-422a,which will provide new targets and new ideas for the treatment of colorectal cancer.
作者
梁鸿
张辉
张超
Liang Hong;Zhang Hui;Zhang Chao(Department of Gastrointestinal Surgery,Henan Provincial People's Hospital,Zhengzhou 450003,China)
出处
《中华放射肿瘤学杂志》
CSCD
北大核心
2020年第12期1118-1123,共6页
Chinese Journal of Radiation Oncology
基金
河南省医学科技攻关计划项目(2018020432)
河南省卫生健康科技英才海外研修工程(HWYX2019116)。
