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Inhibitory effect of microRNA-149-5p in the proliferation and migration of HepG2 and Bel-7402 hepatoma cells
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摘要: 目的明确microRNA-149-5p（miR-149-5p）在肝癌组织中的表达,并探讨其在肝癌细胞中的分子生物学作用。方法收集2010年1月-2014年1月内蒙古医科大学附属医院通过肝切除术获得的65例肝癌组织及相应癌旁组织,采用荧光定量PCR检测65例肝癌组织及相应癌旁组织中miR-149-5p的表达情况。细胞实验分2组,实验组转染miR-149-5p模拟物,对照组转染模拟物阴性对照,采用噻唑蓝比色法和创伤愈合试验检测miR-149-5p对Hep G2和Bel-7402细胞增殖及迁移能力的影响。计量资料组间比较采用t检验或单因素方差分析。结果 miR-149-5p在肝癌组织的表达量（0.14±0.06）明显低于配对的癌旁组织（2.56±0.42）,两者差异有统计学意义（t=7.79,P<0.05）;miR-149-5p在肝癌细胞Hep G2、Bel-7402和正常肝上皮细胞L02中的表达量分别为（1.43±0.25）、（1.77±0.32）和（5.68±0.74）,3组间比较差异有统计学意义（F=11.27,P<0.05）。体外功能实验发现,miR-149-5p模拟物可...Abstract: Objective To investigate the expression of microRNA-149-5p (miR-149-5p) in liver cancer tissue and its molecular and biological role in hepatoma cells. Methods A total of 65 liver cancer tissue samples and corresponding adjacent tissue samples were collected from January 2010 to January 2014, in the Affiliated Hospital of Inner Mongolia Medical University. Quantitative real-time PCR was used to measure the expression of miR-149-5p in liver cancer tissue and corresponding adjacent tissue. The cells were divided into two groups; the cells in the experimental group were transfected with miR-149-5p mimic, and those in the control group were transfected with the negative control of the mimic. MTT colorimetry and wound-healing assay were performed to determine the effect of miR-149-5p on the proliferation and migration of HepG2 and Bel-7402 cells. The t-test or a one-way analysis of variance was used for comparison of continuous data between groups. Results The liver cancer tissue had significantly lower expression of miR-149-5p than the adjacent tissue (0. 14 ± 0. 06 vs 2. 56 ± 0. 42, t = 7. 79, P < 0. 05) . There were significantly differences in the expression of miR-149-5p in Hep G2 (1. 43 ± 0. 25) 、Bel-7402 (1. 77 ± 0. 32) , and the normal hepatic epithelial cells (5. 68 ± 0. 74) (F = 11. 27, P < 0. 05) . The in vitro functional experiment showed that miR-149-5p mimic significantly inhibited the proliferation of 24、48、72 hour of Hep G2 and Bel-7402 hepatoma cells (Hep G2: t = 4. 98, 5. 17, 7. 78, all P < 0. 05; Bel-7402: t = 6. 83, 7. 09, 15. 67, all P < 0. 05) and inhibited the migration of Hep G2 and Bel-7402 hepatoma cells (t = 23. 11, 17. 42, both P < 0. 05) . Conclusion The expression of miR-149-5p is downregulated in liver cancer tissue, and overexpressed miR-149-5p can inhibit the proliferation and migration of hepatoma cells, suggesting that miR-149-5p may be a promising and effective molecular target for the genetic treatment of liver cancer.
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Key words:
- liver neoplasms, experimental /
- microRNAs /
- cell proliferation /
- cell movement
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