miR-181a及miR-138对软骨肉瘤细胞增殖的影响

The Effects of miR-181a and miR-138 on the Proliferation of Chondrosarcoma Cells

【目的】探讨miR-181a及miR-138对软骨肉瘤细胞增殖的影响。【方法】收集20例软骨瘤及20例软骨肉瘤组织标本,采用qRT-PCR检测miR-181a及miR-138表达水平,采用miRNA模拟物促进软骨肉瘤细胞SW1353中miR-181a及miR-138的表达,通过细胞计数法检测miR-181a或miR-138单独过表达及联合过表达对SW1353细胞的生长能力的影响。通过TargetScan数据库预测miR-181a、miR-138与RAS家族关联结构域蛋白8(RASSF8)的结合位点,并采用双荧光素酶报告基因实验进行验证。【结果】与软骨瘤组织相比,miR-181a及miR-138在软骨肉瘤组织中的表达均显著上调(P<0.05);与阴性对照相比,miR-181a及miR-138单独过表达组的细胞生长能力均显著上调(P<0.05),且miR-181a及miR-138联合过表达组细胞生长能力上调最为显著;经预测,miR-181a及miR-138与RASSF8均存在结合位点;与共转染野生型RASSF8+scramble组相比,共转染野生型RASSF8+miR-181a组以及野生型RASSF8+miR-138组的荧光素酶活性均显著降低(P<0.05);与共转染突变型RASSF8+scramble组相比,共转染突变型RASSF8+miR-181a组及突变型RASSF8+miR-138组的荧光素酶活性无显著变化(P>0.05)。【结论】miR-181a及miR-138可能通过抑制RASSF8而促进软骨肉瘤细胞的生长。

【Objective】To investigate the effects of miR-181a and miR-138 on the proliferation of chondrosarcoma cells.【Methods】A total of 20 samples of osteosarcoma and 20 samples of chondrosarcoma tissues were collected.qRT PCR was used to detect the expression levels of miR-181a and miR-138.miRNA mimetics were used to promote the expression of miR-181a and miR-138 in chondrosarcoma cells SW1353.Cell counting was used to detect the effects of miR-181a or miR-138 overexpression alone or in combination on the growth ability of SW1353 cells.The binding sites of miR-181a,miR-138 and RAS family associated domain protein 8(RASSF8)were predicted using the TargetScan database,and gene experiments were validated using dual luciferase reporter technology.【Results】Compared with chondrosarcoma tissue,the expression of miR-181a and miR-138 was significantly upregulated in chondrosarcoma tissue(P<0.05);Compared with the negative control group,the cell growth ability of the miR-181a and miR-138 overexpression groups was significantly increased(P<0.05),and the combined overexpression group of miR-181a and miR-138 showed the most significant upregulation of cell growth ability;According to predictions,both miR-181a and miR-138 have binding sites with RASSF8;Compared with the cotransfected wild-type RASSF8+scaffold group,the luciferase activity in the cotransfected wild-type RASSF8+miR-181a group and wild-type RASSF8+miR-138 group was significantly reduced(P<0.05);Compared with the cotransfected mutant RASSF8+scaffold group,there was no significant change in luciferase activity in the cotransfected mutant RASSF8+miR-181a group and the mutated RASSF8+miR-138 group(P>0.05).【Conclusions】miR-181a and miR-138 may promote the growth of chondrosarcoma cells by inhibiting RASSF8.