摘要
目的研究微小RNA-302a(miRNA-302a)在胃癌标本中的表达水平与临床病理特征的关系,以及其对胃癌细胞的凋亡和细胞周期调控作用,以探讨将miR-302a用于胃癌生物靶向治疗的可能性。方法采用实时PCR(RT-PCR)方法检测胃癌组织和正常胃黏膜组织中miR-302a的相对表达水平,并分析其与对应患者的临床病理资料的相关性。同时,采用RTPCR方法检测miR-302a在胃癌细胞株SGC-7901、MGC-803和BGC-823及正常胃黏膜细胞GES-1中的表达情况;选取miR-302a表达水平最低者为癌细胞,并转染过表达miR-302a的真核重组质粒p CDNA3.1-miR-302a;转染后,采用流式细胞仪Annexin V/PI双染法和PI单染法检测miR-302a对细胞凋亡和细胞周期的影响。结果胃癌组织中miR-302a的相对表达水平显著低于癌旁正常胃癌黏膜组织(P<0.05),且其表达与临床分期、病理分级和转移均有密切关系(P<0.05)。同时,在胃癌细胞SGC-7901、MGC-803和BGC-823中miR-302a的相对表达水平显著低于正常细胞GES-1中的表达(P<0.05),且在BGC-823中表达水平最低。p CDNA3.1-miR-302a质粒转染组与空白组和p CDNA3.1质粒转染组比较,miR-302a质粒转染后48 h后细胞凋亡水平显著升高,且G2/M期细胞数显著增高(P<0.05)。结论在胃癌组织和细胞中miR-302a均呈现低水平表达,当miR-302a过表达后,能显著增高胃癌细胞的凋亡水平并使细胞出现G2/M期阻滞,可能成为胃癌治疗的新靶点。
Objective To investigate the relationship between micro RNA-302a（ miRNA-302a） expression levels and clinical pathological features in gastric carcinoma,and the regulation of gastric cancer cell apoptosis and cell cycle,in order to explore the possibility of miR-302 a to gastric cancer targeted therapy. Methods Real-time RT-PCR method was used to detect the relative expression of miR-302 a in gastric cancer and normal gastric mucosa tissues. The correlation between the clinical pathological data and the clinical pathological data of the patients was analyzed. At the same time,using the real-timeRT-PCR method to detect the expression of miR-302 a in gastric cancer cell lines SGC-7901,MGC-803 and BGC-823 and in normal gastric mucosa cells GES-1,while the expression of miR-302 a was selected for the lowest level of cancer cell,it was transfected by eukaryotic recombinant expression plasmid p CDNA3. 1-miR-302 a miR-302 a. After transfection,flow cytometry Annexin V / PI double staining method and PI staining of miR-302 a were used to examine cell apoptosis and cell cycle effects. Results The relative expression level of miR-302 a in gastric cancer was significantly lower than that in normal gastric cancer（ P〈0. 05）,and its expression was closely related to clinical stage,pathological grade and metastasis（ P〈0. 05）. At the same time,the relative expression level of miR-302 a in gastric cancer cells SGC-7901,MGC-803 and BGC-823 was significantly lower than that in normal cell GES-1（ P〈0. 05）,and the expression level was the lowest in BGC-823. Compared with the blank group and the p CDNA3. 1 plasmid transfection group,the level of apoptosis was significantly increased at 48 hours after miR-302 a transfection,and the cell number in G2 / M phase was significantly increased in p CDNA3. 1-miR-302 a plasmid transfection group（ P〈0. 05）. Conclusion In gastric cancer tissues and cells miR-302 a showed low expression level. With overexpression of miR-302 a,the cell apoptosis can significantly increased and the cells were arrested in G2 / M phase in gastric cancer cells. There may be a new target in the treatment of gastric cancer.
出处
《临床军医杂志》
CAS
2017年第1期42-45,共4页
Clinical Journal of Medical Officers
基金
辽宁省自然科学基金(2013021066)
