摘要
目的 研究miR-135b-5p通过靶向KLF4基因对人胃癌SGC-7901细胞生物学行为的影响。方法通过RT-PCR检测人正常胃黏膜上皮细胞株GES-1及胃癌细胞株SGC-7901、MKN-45、BGC-823、AGS中miR-135b-5p表达水平,将miR-135b-5p inhibitor转染至胃癌SGC-7901细胞中,RT-PCR和Western blotting分别检测转染后细胞中miR-135b-5p和KLF4蛋白表达水平,MTT法、流式细胞术、Transwell实验分别检测转染对细胞增殖、凋亡、侵袭和迁移能力的影响。生物信息学软件预测及双荧光素酶报告基因实验验证KLF4是否为miR-135b-5p的靶基因。结果胃癌细胞SGC-7901、MKN-45、BGC-823、AGS中miR-135b-5p表达水平显著高于人正常胃黏膜上皮细胞GES-1,且胃癌SGC-7901细胞中miR-135b-5p表达水平最高。在SGC-7901细胞中转染miR-135b-5p inhibitor 48 h 后,miR-135b-5p的表达水平显著降低,KLF4 mRNA和蛋白表达水平明显升高。下调miR-135b-5p后SGC-7901细胞增殖能力下降,凋亡率升高,侵袭和迁移能力减弱。双荧光素酶报告基因实验结果显示,KLF4是miR-135b-5p靶基因。结论 miR-135b-5p能够通过靶向KLF4抑制胃癌SGC-7901细胞增殖、侵袭和迁移,诱导细胞凋亡。
Objective To investigate the effect of miR-135 b-5Pon the biological behavior of human gastric cancer SGC-7901 cells by targeting KLF4 gene. Methods The expression of miR-135 b-5Pin human normal gastric mucosal epithelial cell line GES-1 and gastric cancer cell lines SGC-7901, MKN-45, BGC-823 and AGS was detected by RT-PCR. The expressions of miR-135 b-5Pand KLF4 proteins in transfected cells were detected by RT-PCR and Western blotting, respectively. MTT assay, flow cytometry and Transwell assay were used to detect the proliferation of the transfected cells, the effects of apoptosis, invasion and migration. Bioinformatics software prediction and dual luciferase reporter gene experiments verified whether KLF4 was a target gene of miR-135 b-5 p. Results The expressions of miR-135 b-5Pin gastric cancer cells SGC-7901, MKN-45, BGC-823 and AGS were significantly higher than shose in human gastric epithelial cells GES-1, and the expression of miR-135 b-5Pin gastric cancer SGC-7901 cells was the highest. After transfection of miR-135 b-5Pinhibitor for 48 hours in SGC-7901 cells, the expression of miR-135 b-5Pwas significantly decreased, and the expressions of KLF4 mRNA and protein were significantly increased. After down-regulating miR-135 b-5 p, the proliferation of SGC-7901 cells was decreased, the apoptotic rate was increased, and the invasion and migration ability were decreased. The results of the dual luciferase reporter gene assay showed that KLF4 was a miR-135 b-5Ptarget gene. Conclusion miR-135 b-5Pcan inhibit the proliferation, invasion and migration of gastric cancer SGC-7901 cells by targeting KLF4 and can induce apoptosis.
作者
刘小娟
孙科
LIU Xiaojuan;SUN Ke(Department of Gastroenterology,Chengdu Second People’s Hospital,Chengdu 610000,China;Department of Hepatobiliary,Chengdu Second People’s Hospital,Chengdu 610000,China)
出处
《胃肠病学和肝病学杂志》
CAS
2019年第6期614-619,共6页
Chinese Journal of Gastroenterology and Hepatology
基金
广西壮族自治区卫生厅自筹经费科研课题(Z5986879099)
