目的:使用microRNAs基因芯片及实时定量PCR法测定骨肉瘤组织中miR-15a-5p和miR-16-5p的相对表达含量,并与瘤旁组织对比,分析骨肉瘤细胞内miR-15a-5p和miR-16-5p的表达变化.方法:选取34例骨肉瘤组织蜡块样本,使用microRNAs基因芯片观察miR-15a-5p和miR-16-5p在骨肉瘤和瘤旁组织内的表达差异;实时定量PCR法测定骨肉瘤组织和瘤旁组织中miR-15a-5p和miR-16-5p的相对表达含量,并将两种结果对比分析.结果:microRNAs基因芯片结果显示,在骨肉瘤组织中,miR-15a-5p在肿瘤中的表达较瘤旁组织低1.79倍,miR-16-5p较瘤旁组织低1.62倍.实时定量PCR实验结果表明,miR-15a-5p和miR-16-5p表达较瘤旁组织降低,差异有统计学意义(P<0.05).经过统计学计算,miR-15a-5p在肿瘤中的表达较瘤旁组织低3.14倍,miR-16-5p较瘤旁组织低5.65倍.结论:在骨肉瘤中,miR-15a-5p和miR-16-5p表达含量降低,提示这两种microRNAs在骨肉瘤中可能做为抑癌因子存在.
Objective:Use microRNA microarray chip and real time quantitative PCR method to analysis the relative expression levels of miR-15a-5p and miR-16-5p,then compare the difference between osteosarcoma paraffin block samples and peritumoral tissue.Methods:34 cases of osteosarcoma paraffin block samples were chosen,microRNAs microarray chip and real time quantitative PCR method was used to measure the relative expression levels of miR-15a-5p and miR-16-5p.Results:microRNAs microarray chip results showed that the expression level ofmiR-15a-5p (miR-16-5p) in tumor tissue was 1.79 (1.62) times lower than the adjacent tissue.Real time quantitative PCR results showed that compare with tumor adjaccnt tissues,the expression level of miR-15a-5p and miR-16-5p was low in osteosarcoma paraffin block samples.The results were considered significant through statistical analysis (P<0.05).Further analysis results showed that the expression level of miR-15a-5p (miR-16-5p) in tumor tissue was 3.14 (5.65) times lower than the adjacent tissue.Conclusions:miR-15a-5p and miR-16-5p was low expressed in osteosarcoma cells,which suggest that these two microRNAs could be act as oncogene.