长链非编码RNA MALAT1调控骨肉瘤细胞侵袭能力的机制

Mechanism of long-chain non-coding RNA MALAT1 regulating the invasion ability of osteosarcoma cells

目的分析长链非编码RNA人肺腺癌转移相关转录本(MALAT)1与miR-205的关系,研究MALAT1调控骨肉瘤细胞侵袭能力的机制。方法采用实时荧光定量核酸扩增检测(QPCR)对MALAT1、miR-205在两组细胞的表达差异进行比较;利用细胞转染实验分析MALAT1、miR-205的表达关系;采用荧光素酶基因检测分析MALAT1、miR-205的靶向调控关系;利用细胞迁移侵袭试验(Transwell实验)研究MALAT1对骨肉瘤细胞侵袭能力的调控作用。结果MG63、Sao-2细胞的MALAT1的表达水平明显高于hFOB细胞,而MG63、Sao-2细胞的miR-205的表达水平明显低于hFOB细胞(P<0.05)。MG63、Sao-2细胞在转染si-MALAT1后的miR-205表达水平均明显高于对应NC组(P<0.05)。MG63、Sao-2细胞在转染miR-205模拟物后的MALAT1表达水平均明显低于对应NC组(P<0.05)。MG63、Sao-2细胞在共转染miR-205模拟物+MALAT1野生型载体后的荧光值明显低于对应NC组(P<0.05)。MG63、Sao-2细胞在共转染miR-205模拟物+MALAT1突变型载体后的荧光值与对应NG组比较差异无统计学意义(P>0.05)。MALAT1+miR-205模拟物组细胞侵袭数明显高于miR-205模拟物组(P<0.05)。结论在骨肉瘤细胞中,MALAT1的表达较高而miR-205的表达较低,二者的表达呈负相关,且存在相互抑制的关系。MALAT1与miR-205存在靶向调控效应;miR-205可抑制骨肉瘤细胞的侵袭能力,而MALAT1可靶向逆转此抑制效应,促进骨肉瘤细胞的侵袭与进展,对于骨肉瘤的预测与治疗具有重要指导意义。

Objective To analyze the relationship between metastasis-associated lung adenocarcinoma transcript(MALAT)1 and miR-205,and study the mechanism of MALAT1 regulating osteosarcoma cell invasion ability.Methods Real-time Quantitative PCR(QPCR)was used to compare the expression differences of MALAT1 and miR-205 in the two groups of cells.Cellular transfection experiments were used to analyze the expression relationship of MALAT1 and miR-205.The target regulatory relationship between MALAT1 and miR-205 was analyzed by luciferase gene detection;the cell migration invasion test(Transwell test)was used to study the regulatory effect of MALAT1 on the invasion ability of osteosarcoma cells.Results①The expression level of MALAT1 in MG63 and Sao-2 cells was significantly higher than that in hFOB cells,while the expression level of miR-205 in MG63 and Sao-2 cells was significantly lower than that in hFOB cells(P<0.05).②The miR-205 expression level of MG63 and Sao-2 cells after transfection with si-MALAT1 was significantly higher than that of the corresponding NC group(P<0.05).The MALAT1 expression level of MG63 and Sao-2 cells after transfection with miR-205 mimics was significantly lower than that of the corresponding NC group(P<0.05).③The fluorescence value of MG63 and Sao-2 cells after co-transfection of miR-205 mimic+MALAT1 wild-type vector was significantly lower than that of the corresponding NC group(P<0.05).The fluorescence value of MG63 and Sao-2 cells after co-transfection with miR-205 mimic+MALAT1 mutant vector was smaller than that of the corresponding NG group,with no statistical significance(P>0.05).④The number of invasion of MG63 cells after co-transfection of group D(MALAT1+miR-205 mimic)was significantly higher than that of group C(miR-205 mimic)(P<0.05).Conclusions In osteosarcoma cells,the expression of MALAT1 is higher and the expression of miR-205 is lower.The expression of them is negatively correlated,and there is a relationship of mutual inhibition.MALAT1 and miR-205 have targeted regulatory effects;miR-205 could inhibit the invasion ability of osteosarcoma cells,and MALAT1 could be targeted to reverse this inhibitory effect,promote the invasion and progress of osteosarcoma cells,and have a predictive and therapeutic effect on osteosarcoma.