下调HOX转录反义RNA对三阴性乳腺癌细胞MDA-MB-231增殖、周期及侵袭能力的影响

Effects of down-regulation of HOTAIR on proliferation, cycle and invasive ability of triple-negative breast cancer cell line MDA-MB-231

目的研究下调HOX转录反义RNA(HOTAIR)水平对三阴性乳腺癌(TNBC)细胞MDA-MB-231增殖、周期及侵袭能力的影响。方法si-HOTAIR对MDA-MB-231细胞转染,实验分为对照组、si-阴性对照组、si-HOTAIR组,实时荧光定量法测定转染后MDA-MB-231细胞中HOTAIR表达水平;分别用MTT法、流式细胞仪、Transwell小室实验测定转染后MDA-MB-231细胞活性、周期、侵袭能力;蛋白印迹法(WB)测定转染后MDA-MB-231细胞中c-myc、Ki-67、PCNA、基质金属蛋白酶9(MMP-9)、基质金属蛋白酶2(MMP-2)表达水平。结果与MCF-10A细胞组相比,对照组MDA-MB-231细胞中HOTAIR表达水平升高[(1.85±0.21)比(1.06±0.13),P<0.05]。与对照组、si-阴性对照组相比,si-HOTAIR组MDA-MB-231细胞中HOTAIR表达水平(1.34±0.15)、细胞活性(98.04±6.28)%、侵袭能力(35.66±8.04)降低(P<0.05),细胞G0/G1期下调,阻滞在G2/M期;与对照组、si-阴性对照组相比,si-HOTAIR组MDA-MB-231细胞中c-myc、Ki-67、PCNA、MMP-9、MMP-2表达水平降低(P<0.05)。结论HOTAIR在MDA-MB-231细胞中高表达,下调HOTAIR表达水平可抑制MDA-MB-231细胞增殖、阻滞细胞周期、降低肿瘤细胞侵袭能力。

Objective To investigate the effects of down-regulation of HOX transcript antisense RNA(HOTAIR)on proliferation,cell cycle and the invasive ability of triple-negative breast cancer cell line MDA-MB-231.Methods MDA-MB-231 cells were transfected with si-HOTAIR,the experiment was divided into control group,si-negative control group and si-HOTAIR group,the expression of HOTAIR in transfected MDA-MB-231 cells was measured by real-time fluorescence quantitative method;MTT assay,flow cytometry and Transwell cell assay were used to determine the activity,cycle and invasive ability of MDA-MB-231 cells after transfection;and the expression levels of c-myc,Ki-67,PCNA,MMP-9 and MMP-2 in MDA-MB-231 cells were determined by Western blotting(WB).Results Compared with MCF-10A cell group,HOTAIR expression level in MDA-MB-231 cells in control group increased significantly[(1.85±0.21)vs.(1.06±0.13),P<0.05].Compared with control group and si-negative control group,HOTAIR expression level(1.34±0.15),cell activity(98.04±6.28)%and invasive ability(35.66±8.04)of MDA-MB-231 cells in si-HOTAIR group were significantly decreased(P<0.05),G0/G1 cell phase was down-regulated and cells blocked in G2/M phase;compared with the control group and the sinegative control group,the expression levels of c-myc,Ki-67,PCNA,MMP-9 and MMP-2 in MDA-MB-231 cells in the si-HOTAIR group decreased significantly(P<0.05).Conclusion HOTAIR is highly expressed in MDA-MB-231 cells,and downregulation of HOTAIR can inhibit MDA-MB-231 cell proliferation,block cell cycle and reduce cell invasion ability.