摘要
目的:探讨微小RNA-152-3p(miR-152-3p)在结肠癌(CC)中的表达情况及靶向Krüppel样因子4(KLF4)对CC细胞的增殖、迁移和侵袭的影响。方法:通过starBase及TCGA数据库分析预测miR-152-3p与KLF4之间的靶向关系,以及各自在正常组织和CC组织中的表达情况差异;双荧光素酶和RIP实验验证miR-152-3p与KLF4之间的靶向关系;RT-q PCR检测miR-152-3p与KLF4在CC细胞系中的mRNA表达量;Western blot实验检测KLF4在CC细胞系中的蛋白表达水平;MTT实验检测细胞的增殖能力;划痕愈合实验检测细胞的迁移能力;Transwell实验检测细胞的侵袭能力;细胞周期实验和凋亡实验检测细胞的周期分布情况和凋亡百分率。结果:miR-152-3p在CC组织和细胞系中高表达(P<0.001),而KLF4低表达(P<0.01),且miR-152-3p能够靶向下调KLF4的表达(P<0.01)。miR-152-3p过表达能增高CC细胞系的增殖、迁移和侵袭能力,降低细胞周期在G0/G1期的分布,并抑制其凋亡(P<0.05);KLF4过表达可以抑制miR-152-3p对CC细胞增殖、迁移和侵袭能力的促进作用(P<0.05),使G0/G1期细胞比例增加(P<0.05),并促进细胞凋亡(P<0.05)。结论:miR-152-3p通过靶向下调KLF4的表达促进CC细胞的增殖、迁移和侵袭。
Objective:To investigate the expression of microRNA-152-3p(miR-152-3p)in colon cancer(CC)and its effect of targeting Krüppel-like factor 4(KLF4)on proliferation,migration and invasion of CC cells.Methods:The targeting relationship between miR-152-3p and KLF4 was predicted by starBase and TCGA database analysis,as well as the difference in their respective expression in normal and CC tissues.Dual luciferase and RIP experiments verified the targeting relationship between miR-152-3p and KLF4.The mRNA expression of miR-152-3p and KLF4 in CC cell lines was measured by RT-qPCR;the protein expression of KLF4 in CC cell lines was measured by Western blot;the proliferation ability of cells was examined by MTT;the migration ability of cells was examined by scratch healing assay;the invasion ability of cells was examined by Transwell assay;the cell cycle distribution and apoptosis percentage of cells were examined by cell cycle assay and apoptosis assay.Results:miR-152-3p was highly expressed in CC tissues and cell lines(P<0.001),while KLF4 was lowly expressed(P<0.01).miR-152-3p was able to target and downregulate KLF4 expression(P<0.01).miR-152-3p overexpression increased the proliferation,migration and invasion ability of CC cell lines,as well as cell cycle distribution in S and G2/M phases,and inhibited their apoptosis;KLF4 overexpression decreased the proliferation,migration and invasion ability of CC cell lines,contributing to cell cycle retention in G0/G1 phase,and accelerated apoptosis(P<0.05).The up-regulation of KLF4 expression inhibited the promotion of miR-152-3p on the proliferation,migration and invasion of CC cells(P<0.05),increased the proportion of cells in G0/G1 phase(P<0.05),and promoted apoptosis(P<0.05).Conclusion:miR-152-3p promotes proliferation,migration and invasion of CC cells by targeting down-regulation of KLF4 expression.
作者
赵建国
朱晓灵
金学英
江黎明
李振军
陈遐林
ZHAO Jianguo;ZHU Xiaoling;JIN Xueying;JIANG Liming;LI Zhenjun;CHEN Xialin(Department of Oncology,Shaoxing People’s Hospital,Shaoxing Hospital of Zhejiang University,Shaoxing 312000,China;Department of Colorectal Surgery,Shaoxing People’s Hospital,Shaoxing Hospital of Zhejiang University,Shaoxing 312000,China)
出处
《温州医科大学学报》
CAS
2023年第12期954-962,968,共10页
Journal of Wenzhou Medical University
基金
浙江省公益技术研究计划项目(LGF21H160011)。
