CAJ | 학술논문

目的观察沉默叉头框C2 (FOXC2)基因对乳腺癌MDA-MB-231细胞多西紫杉醇化疗敏感性的影响.方法通过脂质体介导将FOXC2短发卡RNA干扰表达质粒FOXC2-小干扰RNA(siRNA)转染至MDA-MB-231细胞,并检测转染48 h后的FOXC2 mRNA及蛋白表达；荧光显微镜观察72 h后的细胞凋亡形态学；流式细胞术检测72 h后的细胞凋亡率及细胞周期.结果与未转染组细胞及空载体组细胞比较,干扰组细胞出现典型凋亡形态学改变,且FOXC2 mRNA及蛋白表达明显降低(P＜0.05)；噻唑蓝(MTT)显示干扰组细胞对多西紫杉醇敏感性增加,半数抑制浓度(IC50)由(6.08±1.23) mg/L降至(0.65 ±0.01) mg/L；同时,干扰组细胞早期凋亡率为(20.01±0.19)％,明显高于其他3组(P＜0.05),且S期及G0/G1期细胞比例减少,G2/M期细胞比例增多(P＜0.05).结论以siRNA下调FOXC2表达可有效诱导乳腺癌MDA-MB-231细胞凋亡,并增强其对多西紫杉醇的化疗敏感性.
목적 관찰침묵차두광C2 (FOXC2)기인대유선암MDA-MB-231세포다서자삼순화료민감성적영향.방법 통과지질체개도장FOXC2단발잡RNA간우표체질립FOXC2-소간우RNA(siRNA)전염지MDA-MB-231세포,병검측전염48 h후적FOXC2 mRNA급단백표체；형광현미경관찰72 h후적세포조망형태학；류식세포술검측72 h후적세포조망솔급세포주기.결과 여미전염조세포급공재체조세포비교,간우조세포출현전형조망형태학개변,차FOXC2 mRNA급단백표체명현강저(P＜0.05)；새서람(MTT)현시간우조세포대다서자삼순민감성증가,반수억제농도(IC50)유(6.08±1.23) mg/L강지(0.65 ±0.01) mg/L；동시,간우조세포조기조망솔위(20.01±0.19)％,명현고우기타3조(P＜0.05),차S기급G0/G1기세포비례감소,G2/M기세포비례증다(P＜0.05).결론 이siRNA하조FOXC2표체가유효유도유선암MDA-MB-231세포조망,병증강기대다서자삼순적화료민감성.
Objective To observe the effects of forkhead box C2 gene (FOXC2) short hairpin RNA on docetaxel chemotherapy sensitivity of human breast cancer MDA-MB-231 cells.Methods MDA-MB-231 cells were transfected with recombinant expression plasmid FOXC2-small interfering RNA (siRNA) by lipofectamine 2000,and the expression of FOXC2 mRNA and protein was detected at 48 h after transfection.The morphological changes of apoptotic cells were observed under fluorescent microscope at 72 h after transfection.Methyl thiazol tetrazolium (MTT) assay was used to examine the inhibitory rate and the 50％ inhibitory concentration (IC50).Flow cytometry was applied to examine apoptosis and cell cycle at 72nd h after transfection.Results As compared with negative control plasmid and FOXC2-siRNA non-transfection group,the typical apoptotic morphology of MDA-MB-231 cells was observed,and the mRNA and protein expression levels of FOXC2 were obviously reduced in FOXC2-siRNA transfection group (P ＜ 0.05).Silencing FOXC2 increased sensitivity of MDA-MB-231 cells to docetaxel (P ＜ 0.05),and IC50 was decreased from (6.08 ±1.23) mg/L to (0.65 ±0.01) mg/L.Meanwhile,the apoptosis rate of MDA-MB-231 cells transfected with pFOXC2-shRNA was (20.01 ± 0.19)％,which was significantly higher than other three groups,and the proportion of cells in S phase and G0/G1 phase was decreased and that in G2/M phase was increased (P ＜0.05).Conclusion Down-regulation of FOXC2 expression by siRNA in breast cancer MDA-MB-231 cells could effectively induce apoptosis of MDA-MB-231 cells and increase their sensitivity to docetaxel.