骨肉瘤miR-96的表达与细胞增殖和凋亡

摘要

背景与目的：微小RNA(microRNA，miRNA)是一类内源性的非编码小分子RNA，其主要通过与靶mRNA作用抑制转录后的翻译。miR-96是在多类肿瘤细胞中高表达的小RNA分子。本研究旨在检测miR-96在骨肉瘤组织中的表达，并研究其在骨肉瘤细胞的增殖、细胞周期及凋亡中的意义。方法：运用TagMan MGB探针法检测40例骨肉瘤组织及对应的骨组织中miR-96的表达；应用反义技术降低骨肉瘤细胞(U2-OS和MG-63)中miR-96的表达；采用MTT比色法检测细胞增殖的改变；应用流式细胞技术检测骨肉瘤细胞细胞周期和凋亡情况。结果：在40例骨肉瘤组织中，62.5%(25/40)的骨肉瘤组织miR-96表达明显高于对应骨组织(P<0.05)；反义miR-96转染骨肉瘤细胞U2-OS和MG-63后，miR-96的表达明显降低，U2-OS和MG-63骨肉瘤细胞生长受到明显抑制，其生长主要停滞在G0/G1期，而S期和G2/M期细胞的比例下降。另外，降低miR-96的表达，U2-OS和MG-63骨肉瘤细胞早期凋亡明显增加。结论：miR-96在骨肉瘤组织中表达明显上调，降低其表达能明显抑制U2-OS和MG-63骨肉瘤细胞的生长，并诱导细胞早期凋亡增加，miR-96可能成为骨肉瘤基因表达调控的新靶点。%　　Background and purpose:MicroRNAs are small non-coding endogenous RNA molecules that can inhibit protein translation partly through binding with target mRNAs. MiR-96 is over expressed in many kinds of human tumor cells. The present study aimed to investigate the expression of miR-96 in osteosarcoma tissues and demonstrate its role in osteosarcoma cell lines proliferation, cell cycle and apoptosis. Methods:The expressions of miR-96 in 40 pairs of osteosarcoma and their adjacent nontumorous tissues were detected by TagMan MGB probe method. The antisense technology was used to decrease the expression of miR-96 in osteosarcoma cells (U2-OS, MG-63), MTT assay and flow cytometry were performed to investigate the effect of miR-96 on the cell proliferation, cell cycles and apoptosis. Results:We examined miR-96 expression levels in 40 pairs of human osteosarcoma and their corresponding nontumorous tissues. miR-96 was found to be over-expressed in 62.5% (25/40) of the osteosarcoma cases (P<0.05). The cell growth ability of U2-OS and MG-63 cells was significantly inhibited following transfection of antisense-microRNA-oligonucleotides-96 (AMO-miR-96). U2-OS and MG-63 cells were mainly detained in G0/G1, and the percentage of cells at S and G2/M decreased. In addition, knocking down the expression of miR-96 in U2-OS and MG-63 cells resulted in an increase in early apoptosis. Conclusion:miR-96 is over-expressed in human osteosarcoma. Inhibition of miR-96 expression can not only effectively suppress the growth of U2-OS and MG-63 cells, but also induce cell apoptosis. MiR-96 may serve as a new molecular target for the regulation of gene expression in osteosarcoma.