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microRNA-10a对胃癌细胞系BGC823迁移和侵袭能力的影响 预览 被引量:11

Effect of microRNA-10a on migration and invasion of gastric cancer cell line BGC823
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摘要 目的:研究人微小RNA-10a(microRNA-10a,miR-10a)对胃癌细胞系BGC823迁移和侵袭能力的影响。方法:利用Transwell小室对胃癌细胞系BGC823进行侵袭筛选,获得高侵袭能力的BGC823-P3亚系;通过miRNA芯片差异分析发现miR-10a在高侵袭能力BGC823-P3细胞的表达显著高于BGC823细胞。通过化学方法合成成熟型的人miR-10a,以脂质体包裹合成的miR-10a(25、50、100、150nmol/L)转染BGC823细胞,并设空白转染、无关序列转染对照组;Real-timePCR分别检测以上各组细胞miR-10a的表达。采用细胞计数试剂盒-8(Cell Counting Kit-8,CCK-8)检测miR-10a对细胞增殖的影响,流式分析检测miR-10a对细胞凋亡的影响,Transwell小室检测miR-10a对细胞的迁移和侵袭能力的影响。结果:化学合成的成熟型miR-10a转染后,BGC823细胞miR-10a表达的提高以100nmol/LmiR-10a转染组最佳,较无关序列转染组提高了2.06倍。miR-10a(100nmol/L)的转染对胃癌细胞系BGC823的增殖和凋亡无明显影响,但对BGC823的迁移和侵袭能力有明显的促进作用,促进率分别为(88.34±0.61)%和(56.02±3.13)%。结论:转染成熟型人miR-10a能使胃癌细胞系BGC823中miR-10a的表达提高,并能显著促进胃癌细胞系BGC823的迁移和侵袭。 Objective:To investigate the effect of human microRNA-10a(miR-10a) on the migration and invasion of gastric cancer cell line BGC823. Methods: The Transwell system was used to select highly invasive sub-cell lines from gastric cancer cell line BGC823. Using miRNA microchip, we compared the miRNA expression in paired cell lines with high and low invasive potentials. MiR-10a was relatively overexpressed in the highly invasive cell lines when compared with its counterpart. The mature type human miR-10a was synthesized chemically. The synthesized miR-10a (25, 50, 100, and 150 nmol/L) was transfected into BGC823 cells via lipofectamin 2000. Cells were also transfected with empty vectors and unrelated fragment to serve as controls. The expression of mature type miR-10a was detected by real-time PCR. Cell counting kit- 8 was used to study the effect of miR-10a on the proliferation of BGC823 cells. Flow cytometry was performed to detect the effect of miR-10a on the apoptosis of BGC823 ceils. The migration and invasion of BGC823 cells were investigated by Transwell assay. Results: Real-time PCR showed that cells transfected with mature type miR-10a had significantly higher expression of miR-10a, with the optimal concentration of miR-10a being 100 nmol/L; the associated miR-10a expression was 2.06 folds that of unrelated group, miR-10a ( 100 nmol/L) had no obvious influence on the proliferation and apoptosis of BGC823 cells; however, it promoted the migration and invasion of BGC823 ceils, with the promoting rates being (88.34±0.61 )% and (56.02 ± 3. 13 )%, respectively. Conclusion: Synthesized mature type human miR-10a can effectively enhance miR-10a expression and promote the migration and invasion of the BGC823 cells.
作者 彭亮 潘健 胡海 孙力超 周转 冉宇靓 杨治华 PENG Liang, PAN Jian, HU Hai, SUN Li-ehao, ZHOU Zhuan, RAN Yu-liang , YANG Zhi-hua (Department of Cellular Biology and Molecular Biology, Cancer Institute, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100021, China)
出处 《中国肿瘤生物治疗杂志》 CAS CSCD 2008年第5期 417-421,共5页 Chinese Journal of Cancer Biotherapy
基金 国家自然科学基金资助项目(No.30570818) 国家重点基础研究发展计划(973)资助项目(No.2009CB521804).
关键词 胃癌细胞 微小RNA-10a 迁移 侵袭 gastric cancer cell microRNA-10a migration invasion
作者简介 彭亮(1980-),男,河北省唐山市人,博士,主要从事肿瘤基因功能方面的研究 Corresponding author. E-mail: ran_yuliang@ 126. com
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