摘要
目的研究SOX1过表达对Hela细胞凋亡、细胞周期以及Wnt通路的影响。方法按照实验室方法进行Hela细胞培养及转染,分为两组:对照组为空质粒转染组,实验组为SOX1转染组,采用荧光实时定量聚合酶链反应(Q-RT-PCR)法检测Hela细胞中SOX1 mRNA表达;流式细胞术检测过表达SOX1对Hela细胞凋亡及细胞周期的影响;Western blot检测Wnt通路相关蛋白包括SOX1、Wnt5a、CCND1、GSK-3B、β-catenin和GAPDH蛋白表达情况。结果Q-RT-PCR检测结果显示:与对照组比较,实验组SOX1 mRNA表达显著升高(t=2.16,P=0.003735);流式细胞术检测结果发现:与对照组相比,实验组的细胞凋亡数目明显增多,差异有统计学显著性(P<0.05),SOX1过表达后,G0/G1期细胞比例增加,G2/M期细胞比例减少,差异均具有统计学意义(P<0.05);Western blot检测结果显示:与对照组相比,SOX1过表达Hela细胞中SOX1、Wnt5a、CCND1、β-catenin的蛋白表达水平均明显增加(P<0.05),而GSK-3B和GAPDH蛋白表达无显著差异(P>0.05)。结论过表达SOX1通过调控Wnt通路,促进Hela细胞凋亡并抑制细胞周期进程,发挥抑癌作用。
Objective To investigate the effects of SOX1 overexpression on Hela cell apoptosis,cell cycle and Wnt pathway.Methods Hela cells were cultured and transfected according to the laboratory methods and divided into two groups:control group was empty plasmid transfection group,experimental group was SOX1 transfection group.The expression of SOX1 mRNA in Hela cells was detected by Q-RT-PCR.The effects of SOX1 overexpression on Hela cell apoptosis and cell cycle were detected by flow cytometry.Western blot was used to detect Wnt pathway related proteins,including SOX1,Wnt5 a,CCND1,GSK-3 B,β-catenin and GAPDH protein.Results Q-RT-PCR results showed that the expression of SOX1 mRNA in experimental group was significantly higher than that in control group(t=2.16,P=0.003735);The results of flow cytometry showed that,compared with the control group,the number of apoptotic cells in the experimental group increased significantly(P<0.05).After SOX1 overexpression,the proportion of cells in G0/G1 phase increased,while the proportion of cells in G2/M phase decreased(P<0.05);Western blot results showed that compared with the control group,the expression of SOX1,Wnt5 a,CCND1,β-catenin in Hela cells were significantly increased(P<0.05),but there was no significant difference in the expression of GSK-3 B and GAPDH(P>0.05).Conclusion Overexpression of SOX1 plays an important role in tumor inhibition by regulating Wnt pathway,promoting Hela cell apoptosis and inhibiting cell cycle progression.
作者
袁敏
姚立丽
古扎丽努尔·阿不力孜
YUAN Min;YAO Li-li;Guzhalinuer·Abulizi(Department of Gynecology,Affiliated Tumour Hospital of Xinjiang Medical University,Ulumuqi 830011,China)
出处
《哈尔滨医科大学学报》
CAS
2021年第5期466-470,共5页
Journal of Harbin Medical University
基金
省部共建中亚高发病成因与防治国家重点实验室开放课题(SKL-HIDCA-2020-34)。
