文章摘要
张荣,姚春和.过表达微小 RNA-877-5p通过靶向叉头框转录因子 M1调控胃癌细胞活力和凋亡[J].安徽医药,2022,26(10):2026-2032.
过表达微小 RNA-877-5p通过靶向叉头框转录因子 M1调控胃癌细胞活力和凋亡
Over-expression of miR-877-5p regulates the viability and apoptosis of gastric cancer cells by targeting FOXM1
  
DOI:10.3969/j.issn.1009-6469.2022.10.028
中文关键词: 胃肿瘤  叉头转录因子类  叉头框蛋白 M1  微小 RNA-877-5p  细胞活力  凋亡
英文关键词: Stomach neoplasms  Forkhead transcription factors  Forkhead box protein M1  miR-877-5p  Cell viability  Apoptosis
基金项目:
作者单位E-mail
张荣 延安大学咸阳医院消化内科二病区陕西咸阳 712000  
姚春和 延安大学咸阳医院普通外科一病区陕西咸阳 712000 wts0s2@163.com 
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中文摘要:
      目的探讨微小 RNA-877-5p(miR-877-5p)对胃癌细胞活力、凋亡的影响及其分子机制。方法本研究时间为 2020年 1—7月。胃癌和正常胃黏膜上皮细胞株购自美国典型培养物保藏中心。实时定量基因扩增荧光检测( qPCR)检测胃癌细胞株 HGC-27、SUN-1、AGS和正常胃黏膜上皮细胞株 GES-1中 miR-877-5p和叉头框转录因子 M1(FOXM1)信使核糖核酸( mRNA)表达。建立 miR-877-5p过表达或抑制 FOXM1表达细胞株,观察其在 HGC-27细胞的活力、凋亡中的作用。 MTT法检测细胞的活力,流式细胞术检测细胞凋亡,蛋白质印迹法( Western blotting)检测 FOXM1、细胞周期蛋白 D1(cyclinD1)、细胞周期依赖性激酶抑制因子 p21、p27、B细胞淋巴瘤 /白血病 -2(Bcl-2)、 Bcl-2相关 X蛋白( Bax)、活化的含半胱氨酸的天冬氨酸蛋白水解酶 3(cleaved-caspase 3)蛋白表达。 TargetScan预测结合双荧光素酶报告实验分析 miR-877-5p和 FOXM1的靶向关系。共转染 miR-877-5p模拟物和 FOXM1过表达载体( pcDNA-FOXM1),研究 FOXM1过表达对 miR-877-5p过表达诱导的 HGC-27细胞增殖和凋亡的影响。结果与正常胃黏膜上皮细胞 GES-1比较,胃癌细胞 HGC-27、SUN-1、AGS中的 miR-877-5p表达下调( 1.00±0.08比 0.34±0.03,0.51±0.05,0.44±0.04,P<0.05)FOXM1 mRNA和蛋白表达上调( 1.00±0.09比 2.41±0.23,2.58±0.24,2.26±0.23,P< 0.05)。 miR-877-5p过表达显著降低 HGC-27细,胞 48 h、72 h的细胞活力( P<0.05),明显提高细胞凋亡率、 p21、p27、Bax、cleaved-caspase3蛋白的水平( P<0.05)显著减少 cyclinD1、Bcl-2蛋白表达量( P<0.05)。抑制 FOXM1表达显著降低 48 h、72 h的细胞活力( P<0.05)提高细胞凋亡率、 p,21、Bax蛋白水平( P<0.05),减少 cyclinD1、Bcl-2蛋白表达量( P<0.05)。 miR-877-5p靶向调控 FOXM1的表达,。FOXM1过表达后, miR-877-5p过表达对 HGC-27细胞增殖、 cyclinD1、Bcl-2蛋白表达的抑制作用被逆转,其对细胞凋亡、 p21、Bax蛋白表达的促进作用也被逆转。结论过表达 miR-877-5p通过靶向调控 FOXM1表达抑制胃癌细胞的活
英文摘要:
      Objective To investigate the effect of microRNA-877-5p (miR-877-5p) on the viability and apoptosis of gastric cancer cells and its molecular mechanism.Methods This study was conducted from January to July 2020. Gastric cancer and normal gastricmucosal epithelial cell lines were purchased from the American Type Culture Collection. qPCR was used to detect the expressions ofmiR-877-5p and Forkhead box M1 (FOXM1) transcription factor messenger ribonucleic acid (mRNA) in gastric cancer cell lines HGC-27, SUN-1, AGS and normal gastric mucosal epithelial cell line GES-1. The miR-877-5p over-expressing or inhibiting FOXM1 express. ing cell line was established, and their role in the proliferation and apoptosis of HGC-27 cells was observed. The cell viability was de.tected by MTT assay, apoptosis was detected by flow cytometry, and the protein levels of FOXM1, cyclinD1, cycle-dependent kinase in. hibitor (p21, p27), B cell lymphoma/leukemia-2 (Bcl-2), Bcl-2 associated X protein (Bax), cleaved cysteinyl aspartate specific protein. ase 3 (cleaved-caspase 3) were determined by Western blotting. TargetScan together with the dual luciferase reporter assay was used toanalyze the targeting relationship between miR-877-5p and FOXM1. miR-877-5p mimic and FOXM1 overexpression vector (pcDNA-FOXM1) were co-transfected to study the effect of FOXM1 over-expression on the viability and apoptosis of HGC-27 cells induced by miR-877-5p over-expression.Results Compared with normal gastric mucosal epithelial cell line GES-1, the expressions of miR-877-5p in the gastric cancer cell lines HGC-27, SUN-1 and AGS were down-regulated (1.00±0.08 vs. 0.34±0.03, 0.51±0.05, 0.44±0.04, P< 0.05), and the FOXM1 mRNA and protein levels were up-regulated (1.00±0.09 vs. 2.41±0.23, 2.58±0.24, 2.26±0.23, P<0.05). Over-ex. pression of miR-877-5p significantly decreased the cell viability of HGC-27 cells at 48 h and 72 h (P<0.05), evidently increased the apoptotic rate, p21, p27, Bax, cleaved-caspase3 protein levels (P<0.05), and reduced the levels of cyclinD1 and Bcl-2 protein (P<0.05). Inhibition of FOXM1 expression obviously reduced cell viability at 48h and 72h (P<0.05), increased apoptotic rate, p21 and Bax pro. tein levels (P<0.05), and decreased cyclinD1 and Bcl-2 protein levels (P<0.05). miR-877-5p targeted to regulate the expression of FOXM1. Over-expression of FOXM1 reversed the inhibitory effect of miR-877-5p over-expression on the proliferation of HGC-27 cells, the protein expressions of cyclinD1 and Bcl-2, and the promotion of apoptosis, p21 and Bax protein levels.Conclusion Over-expres. sion of miR-877-5p inhibits the viability of gastric cancer cells and induces apoptosis by targeting FOXM1 expression.
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