摘要
目的检测微小RNA-3（miR-93）在胃癌组织、正常胃黏膜组织及胃癌细胞株中的表达，分析其表达与胃癌临床病理学特征的相关性；在体外研究miRN3对胃癌细胞增殖功能的影响及其相关的调节机制。方法实时定量反转录聚合酶链反应（RT-qPCR）被用来检测胃癌组织、正常胃黏膜组织及胃癌细胞株中miRN3的表达量。细胞计数试剂盒（CCK-8）、流式细胞术及Western blot法分别被用来研究miR-93对胃癌细胞增殖、周期分布的影响及其介导胃癌细胞增殖相关的调节机制。结果在胃癌组织中，miR-93表达量显著升高（癌旁组织中表达量为0．86±0．09，而胃癌组织中的表达量为2．08±0．13，P〈0．01），异常表达的miR-93与肿瘤的临床病理学分期相关（P〈0．01）；此外，miR03在胃癌细胞株AGS、HTB135及MKN-74中的水平分别为2．05、3．63、3．17，显著高于其在胃黏膜正常细胞株GES-1中的水平（0．96，P〈0．01）；在体外，miR-93可通过负调控细胞周期依赖性蛋白激酶抑制因子1A（p21）的表达而发挥促肿瘤效应：在转染miR03后48、72、96h，AGS细胞的增殖能力显著增强（P〈0．05）；此外，转染miR-93可促进AGS细胞周期的演进（G0／G1期细胞由56．37％下降至40．89％，而S期细胞由32．58％上升至45．27％）；相反，miR-93抑制剂却能显著抑制AGS细胞的增殖能力（P〈0．05），诱导细胞周期阻滞在G0／G1期（G0／G1期细胞由56．37％上升至63．78％，而S期细胞由32．58％下降至20．53％，P〈0．05）。结论检测miR-93有助于诊断早期胃癌，抑制其表达可为胃癌的治疗提供参考依据。
Objective To explore microRNA- 93 （miR- 93） expression and its role in gastric cancer （GC）. Methods MiR- 93 levels were detected by realtime reverse transcriptase -polymerase chain reaction （RT - qPCR） in 50 paired GC samples, 10 normal gastric mucosas, and three GC cell lines. The correlation between dysregulated miR -93 and clinicopathological features of GC patients was analyzed. The effects of miR -93 on GC cell proliferation and cell cycle distribution were examined by cell counting kit -8 （CCK -8 ） and flow cytometry. Western blotting was applied to explore the target gene of miR -93 in GC cells. Results As compared with their normal counterparts, miR -93 expression was obviously overexpressed in GC tissues （0. 86 ± 0. 09 vs. 2. 08 ±0. 13 ,P 〈0. 01 ） and three GC cell lines （0.96 vs. 2.05, 3.65, 5.17, respectively,P 〈0. 01 ）, and its levels in GC tissues was positively correlated with tumor clinical stages （ P 〈 0. 01 ）. miR - 93 may act an oncogene through negatively regulating p21 expression ： enforced expression of miR - 93 significantly prompted AGS cell proliferation rate （ P 〈 0. 05 ） and its cell cycle progression （ the percentage of G0/G1 cells was decreased from 56.37% to 40. 89% , while the portion of S phase cells was increased from 32. 58% to 45.27% ,P 〈0. 05）. On the contrary, miR - 93 inhibitor restrained AGS cell proliferation rate （ P 〈 0. 05 ） and cell cycle arrest at G0/G1 （ compared with control group, the number of cells at G0/G1 was greatly increased from 56. 37% to 63.78% , whereas the number of cells at S phase obviously diminished from 32. 58% to 20. 53% ,P 〈0. 05 ）. Conelusion Our data suggest that miR - 93 may act as a promising diagnostic candidate for early - stage GC and it may provide a novel approach for controlling GC development.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第2期299-302,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金（81172294）
