微小核糖核酸-218在人骨肉瘤组织及Saos-2细胞中的表达及其意义

Expression level and significance of miRNA-218 in human osteosarcoma and Saos-2 cells

目的:检测微小核糖核酸-218(miRNA-218,miR-218)在人类骨肉瘤组织和骨肉瘤细胞Saos-2中的表达水平并探讨miR-218对骨肉瘤发生发展的影响。方法:收集12例(对)人类骨肉瘤及其癌旁组织(正常对照)样本,培养人骨肉瘤细胞系Saos-2,并建立高表达miR-218的细胞系Saos-2miR-218+,运用实时定量PCR方法检测上述组织及细胞中miR-218的表达水平,通过ATPlite化学发光法检测细胞增殖水平,趋化小室实验观察高表达miR-218的Saos-2细胞的侵袭能力变化。结果:与正常对照组相比较,miR-218在骨肉瘤组织中的表达显著下降(0.61±0.02vs 0.37±0.03),Saos-2细胞中miR-218的表达亦显著下调(0.59±0.15 vs 0.17±0.04);Saos-2miR-218+细胞增殖水平较对照细胞及空载体转染细胞显著降低[培养72 h的活细胞数分别为(1200±85)、(1960±41)、(1950±52)个/μL],以上差异均有统计学意义(P<0.05);趋化小室实验显示,Saos-2miR-218+细胞的侵袭能力下降。结论:miRNA-218在人骨肉瘤组织及Saos-2细胞中的表达水平下降,提高miRNA-218的表达水平可抑制Saos-2细胞增殖和侵袭。

Objective:To detect the expression level of miRNA-218(miR-218)in human osteosarcoma tissues and Saos-2 cells,and explore the effect of miR-218 on the development of osteosarcoma.Methods:Twelve cases of human osteosarcoma and their adjacent normal tissues(normal control)were collect-ed,human osteosarcoma cell line Saos-2 was cultured,and a cell line Saos-2 miR-218+with high expres-sion of miR-218 was established.The expression levels of miR-218+in the above tissues and cells were detected using real-time quantitative polymerase chain reaction(PCR).ATPlite luminescence as-say systems was used to detect the cell proliferation level,and the chemotactic chamber experiment was used to observe the invasion ability of Saos-2 cells with high expression of miR-218.Results:Compared with that in normal control,the expression of miR-218 in osteosarcoma was significantly decreased(0.61±0.02 vs 0.37±0.03),and that in Saos-2 cells was also significantly down-regulated(0.59±0.15 vs 0.17±0.04).The proliferation level of Saos-2 miR-218+cell was significantly lower than that of control cells and empty vector transfected cells(living cells count at 72 h was 1200±85,1960±41,and 1950±52 per micro-liter,respectively),with significant difference(all P<0.05).The chemotactic chamber experiment showed that the invasiveness of Saos-2 miR-218+cells decreased.Conclusion:The expression of miR-218 in human osteosarcoma tissue and Saos-2 cells decreased,and higher expression of miR-218 could inhibit the proliferation and invasion of Saos-2 cells.