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.
Chinese Journal of Cancer Biotherapy
Corresponding author. E-mail： ran_yuliang@ 126. com