摘要
目的探究下调XRCC6对人结直肠癌细胞凋亡、侵袭的影响及其与ATM/CHK1/p53信号通路的相关性。方法将人结直肠癌细胞SW620分为对照组、si⁃NC组和si⁃XRCC6组。MTT法检测各组细胞活力;流式细胞法检测各组细胞凋亡水平;细胞划痕实验检测各组细胞迁移能力;Transwell检测各组细胞侵袭能力;定量聚合酶链反应(qPCR)检测ATM、CHK2、p53的转录水平;免疫印迹(Westernblot)检测ATM、CHK2、p53的蛋白表达和磷酸化(p⁃ATM、p⁃CHK2、p⁃p53)水平。结果MTT实验结果表明,与对照组和si⁃NC组相比,si⁃XRCC6组细胞活力显著降低(P<0.05);流式细胞实验结果表明,与对照组和si⁃NC组相比,si⁃XRCC6组细胞存活比例降低(P<0.01),早期凋亡比例增加(P<0.01),晚期凋亡细胞比例显著增加(P<0.01),si⁃NC组和si⁃XRCC6组差异无统计学意义(P>0.05);细胞划痕及Transwell实验结果表明,与对照组和si⁃NC组相比,si⁃XRCC6组细胞迁移能力降低(P<0.01),侵袭能力降低(P<0.01)。qPCR及Westernblot结果表明,与对照组和si⁃NC组相比,si⁃XRCC6组细胞ATM、CHK2、p53的转录和蛋白表达水平有上升的趋势,其磷酸化蛋白p⁃ATM、p⁃CHK2、p⁃p53表达水平也具有上升的趋势。结论下调XRCC6可以抑制人结直肠癌细胞活性,诱导细胞早期凋亡,其机制可能与ATM、CHK2、p53信号通路的调控及其磷酸化水平的调控相关。
Objective To explore the effect of down⁃regulation of XRCC6 on apoptosis and invasion of colorectal cancer cells and the changes of ATM/CHK2/p53 signaling pathway.Methods Human colorectal cancer SW620 cells were divided into three groups:control group,si⁃NC group and si⁃XRCC6 group.MTT assay was used to detect cell viability.Flow cytometry was used to detect cell apoptosis.Cell scratch test was used to detect cell migration.Transwell was used to detect cell invasion.qPCR was used to detect ATM,CHK2 and p53 mRNA expression.Western blot was used to detect ATM,CHK2,p53 protein expression and the phosphorylation level of p⁃ATM,p⁃CHK2 and p⁃p53.Results MTT results showed that the cell viability of si⁃XRCC6 group was significantly lower than that of control group and si⁃NC group(P<0.05).Flow cytometry results showed that the survival rate of si⁃XRCC6 group was lower than that of control group and si⁃NC group(P<0.01),the early apoptosis rate was increased(P<0.01),the proportion of late apoptotic cells increased significantly(P<0.01),and there was no significant difference between si⁃NC group and si⁃XRCC6 group(P>0.05).Compared with the control group and si⁃NC group,the migration ability and invasion ability of si⁃XRCC6 group decreased(P<0.01).qPCR and Western blot results showed that the mRNA and protein expression levels of ATM,CHK2 and p53 in si⁃XRCC6 cells increased,and the phosphorylation level of p⁃ATM,p⁃CHK2 and p⁃p53 also increased.Conclusion Down regulation of XRCC6 can inhibit the activity of human colorectal cancer cells and induce early apoptosis,which may be related to the regulation of ATM,CHK2 and p53 signaling pathways and their phosphorylation levels.
作者
刘建通
方育
高洪波
宋蒙蒙
林超
Liu Jiantong;Fang Yu;Gao Hongbo;Song Mengmeng;Lin Chao(Department of General Surgery,Beijing Nuclear Industry Hospital,Beijing 102413,China;Department of General Surgery,Xuanwu Hospital,Capital Medical University,Beijing 100053,China;Department of Radionuclide Treatment Center,Beijing Nuclear Industry Hospital,Beijing 102413,China)
出处
《肿瘤代谢与营养电子杂志》
2021年第6期615-619,共5页
Electronic Journal of Metabolism and Nutrition of Cancer
基金
北京市卫生健康委员会科研项目(2019J500)。
关键词
XRCC6
人结直肠癌细胞
凋亡
侵袭
磷酸化
X⁃ray repair cross complementing 6
Human colorectal cancer cells
Apoptosis
Attack
Phosphorylation
