FAM21对绒毛膜癌细胞系JEG-3迁移和侵袭功能的影响

Influence of FAM21 on migration and invasion properties of the choriocarcinoma cell line JEG-3

目的探讨FAM21对人绒毛膜癌细胞系JEG-3迁移和侵袭功能的影响。方法设计并合成2条FAM21特异性小分子干扰RNA(siRNA 1和siRNA 2)及1条随机序列siRNA(control siRNA),瞬时转染JEG-3细胞,依次设为siRNA1组、siRNA 2组和control siRNA组。同时选取未行转染的JEG-3细胞作空白对照(空白组)。普通PCR法检测正常胎盘滋养细胞株HTR-8/Sveno和JEG-3细胞中FAM21 mRNA水平。分别于转染后24、48h采用实时荧光定量PCR和Western blotting检测各组FAM21的mRNA和蛋白水平,Transwell法检测各组转染24h后的细胞迁移和侵袭能力,实时荧光定量PCR检测各组转染24h后的迁移侵袭相关基因(KISS-1、BRMS1和INSL4)的mRNA水平。结果 JEG-3细胞的FAM21 mRNA水平高于HTR-8/Svneo细胞(P<0.05)。siRNA 1组和siRNA 2组转染24h后的FAM21 mRNA和转染48h后的FAM21蛋白水平均低于control siRNA组和空白组(P<0.05),转染24h后的迁移和侵袭能力均低于control siRNA组和空白组(P<0.05)。siRNA 1组和siRNA 2组转染24h后的KISS-1和BRMS1 mRNA水平降低,而INSL4 mRNA水平升高,与其余两组比较,差异均有统计学意义(P<0.05);control siRNA组和空白组上述指标的差异均无统计学意义(P>0.05)。结论 FAM21在人绒毛膜癌JEG-3细胞中具有调节迁移和侵袭的功能,在绒毛膜癌的病理机制中发挥了重要的作用。

Objective To explore the influence of FAM21 on migration and invasion properties of choriocarcinoma cell line JEG-3. Methods Two FAM21-specific small interfering RNA（ siRNA1 and siRNA2） and one scramble siRNA（ control siRNA） were designed. According to the experimental design, the JEG-3 cells with stable transfection of siRNA1, siRNA2 and control siRNA were assigned into three groups：siRNA1 group, siRNA2 group and control siRNA group. The JEG-3 cells without transfection were selected as blank（ blank group） . FAM21 mRNA levels of JEG-3 cells and human trophoblast HTR-8/Svneo cells were detected by normal PCR. The quantitative PCR and Western blotting were employed to measure the FAM21 mRNA level at 24h after transfection and protein lev-el at 48h after transfection of each group, respectively. The migration and invasion properties of each group at 24h after transfection were detected by Transwell method. The quantitative PCR was employed to detect the mRNA levels of invasion-related genes（ KISS-1, BRMS1 and INSL4） at 24h after transfection. Results There was a higher mRNA level of FAM21 in JEG-3 cells versus HTR-8/Svneo cells （ P＆lt;0？05） . The FAM21 mRNA level at 24h and FAM21 protein level at 48h after transfection of siRNA1 and siRNA2 groups were lower than those of control siRNA group and blank group with significant differences（ P＆lt;0？05） . There were lower migration and invasion properties in siRNA1 and siRNA2 groups versus control siRNA group and blank group at 24h after transfection（P＆lt;0？05）. There were decreased mRNA levels of KISS-1 and BRMS1 and increased level of INSL4 in siRNA1 and siRNA2 groups versus control siRNA group and blank group at 24h after transfection（P＆lt;0？05）. No significant difference was observed on the above indicators be-tween control siRNA group and blank group（ P＆gt;0？05） . Conclusion FAM21 protein can regulate the migration and invasion properties of the choriocarcinoma cell line JEG-3, which plays an important role in the pathogenesis of choriocarcinoma.