miRNA-130b在骨肉瘤组织中的表达及其对骨肉瘤细胞增殖与凋亡的影响

发布时间：2018-07-23 15:23

【摘要】：目的探讨骨肉瘤组织中miRNA-130b的表达及其对骨肉瘤细胞增殖、细胞周期以及凋亡的影响。方法用Real-time PCR法检测48例骨肉瘤患者肿瘤组织与癌旁组织中miRNA-130b的表达水平。用脂质体法将miRNA-130b inhibitor瞬时转染入U2人骨肉瘤细胞,实时定量RT-PCR检测U2细胞miRNA-130b的表达,CCK8增殖实验检测各组细胞的增殖情况,流式细胞仪检测各组细胞周期及细胞凋亡率,荧光素酶基因报告实验检测miRNA-130b与RUNX3之间的调控机制、荧光素酶的相对活性和转染后RUNX3及miRNA-130b的表达水平。结果骨肉瘤组织中miRNA-130b的表达明显高于癌旁组织,转染miRNA-130b inhibitor后U2细胞miRNA-130b的表达水平明显受抑制,人骨肉瘤细胞U2转染miRNA-130b inhibitor的细胞增殖速度明显下降,细胞周期阻滞在G0/G1期,凋亡率增高,细胞中RUNX3蛋白及m RNA表达水平明显升高;miRNA-130b显著抑制野生型RUNX3-3’UTR表达载体的荧光素酶活性;抑制miRNA-130b表达后,RUNX3表达明显升高;抑制RUNX3表达后,miRNA-130的表达无明显变化。结论 miRNA-130b在骨肉瘤中呈高表达水平,抑制miRNA-130b可能通过调控RUNX3抑制骨肉瘤细胞的增殖,并促进细胞凋亡。
[Abstract]:Objective to investigate the expression of miRNA-130b in osteosarcoma and its effect on proliferation, cell cycle and apoptosis of osteosarcoma cells. Methods Real-time PCR method was used to detect the expression of miRNA-130b in 48 patients with osteosarcoma. MiRNA-130b inhibitor was transiently transfected into U2 human osteosarcoma cells by liposome method. The expression of miRNA-130b in U2 cells was detected by real-time quantitative RT-PCR. The proliferation of U2 cells was detected by CCK8 proliferation assay. The cell cycle and apoptosis rate of each group were detected by flow cytometry. The regulatory mechanism between miRNA-130b and RUNX3, the relative activity of luciferase and the expression level of RUNX3 and miRNA-130b after transfection were detected by luciferase gene report experiment. Results the expression of miRNA-130b in osteosarcoma tissue was significantly higher than that in paracancerous tissue. The expression of miRNA-130b in U2 cells was significantly inhibited after transfection of miRNA-130b inhibitor. The proliferation rate of U2 transfected miRNA-130b inhibitor cells in human osteosarcoma cells decreased significantly, and the cell cycle was blocked in G0/G1 phase. The expression of RUNX3 protein and m RNA in the cells increased significantly with the increase of apoptosis rate, the luciferase activity of wild-type RUNX3-3'UTR expression vector was significantly inhibited by miRNA-130b, the expression of rUNX3 increased significantly after inhibiting miRNA-130b expression, and the expression of miRNA-130 did not change after inhibiting RUNX3 expression. Conclusion miRNA-130b is highly expressed in osteosarcoma. Inhibition of miRNA-130b may inhibit the proliferation of osteosarcoma cells and promote apoptosis by regulating RUNX3.
【作者单位】：新余钢铁集团有限公司中心医院骨科;南昌大学第二附属医院骨科;
【分类号】：R738.1

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