沉默MACC1基因对人卵巢癌耐药细胞增殖、凋亡和侵袭的影响

发布时间：2018-03-26 19:32

本文选题：卵巢癌　切入点：结肠癌转移相关基因　出处：《山东医药》2017年26期

【摘要】：目的观察应用基因沉默技术抑制人卵巢癌耐药细胞结肠癌转移相关基因1(MACC1)表达后,细胞增殖、凋亡和侵袭能力的变化及其分子机制。方法将人卵巢癌耐药细胞株SKOV-3/DDP分成空质粒组和转染组,分别转染空质粒p-super-EGFP、重组质粒p-super-EGFP-MACC1 shRNA,另取未经处理的SKOV-3/DDP细胞作为空白对照组。采用RT-PCR方法检测各组MACC1 mRNA表达,Western blotting法检测MACC1蛋白表达,MTT法检测细胞增殖能力,流式细胞术检测细胞凋亡情况,体外黏附试验检测细胞体外黏附力,Transwell小室检测细胞侵袭能力,Western blotting法测定C-met、ERK1/2、p-ERK1/2蛋白表达。结果与空白对照组和空质粒组比较,转染组MACC1 mRNA和蛋白表达减少,细胞48、72 h增殖能力减弱,细胞凋亡率增加,体外黏附和侵袭能力减弱,C-met、pERK1/2蛋白表达增加(P均0.05),而空白对照组和空质粒组各指标差异无统计学意义(P均0.05)。结论MACC1基因表达抑制后,SKOV-3/DDP的增殖、黏附和侵袭能力减弱,细胞凋亡增加,这可能与HGF/C-met和ERK通路受抑制有关。
[Abstract]:Objective to observe the effect of gene silencing on the proliferation of human ovarian cancer cells after inhibiting the expression of metastasis associated gene 1 (MACC1) in human ovarian cancer cells. Methods Human ovarian cancer cell line SKOV-3/DDP was divided into empty plasmid group and transfection group. The empty plasmids p-super-EGFP were transfected, the recombinant plasmid p-super-EGFP-MACC1 shRNAs were transfected, and the untreated SKOV-3/DDP cells were used as the blank control group. The expression of MACC1 mRNA in each group was detected by RT-PCR method and the expression of MACC1 protein was detected by Western blotting method. Flow cytometry was used to detect cell apoptosis, and in vitro adhesion assay was used to detect cell invasion ability. Western blotting assay was used to detect the expression of C-metERK1 / 2 p-ERK1 / 2 protein. The results were compared with those of blank control group and empty plasmid group. In the transfected group, the expression of MACC1 mRNA and protein decreased, the proliferation ability of the cells decreased and the apoptosis rate increased. The ability of adhesion and invasion was decreased in vitro. The expression of pERK1 / 2 protein increased significantly (P < 0.05), but there was no significant difference between blank control group and blank plasmid group (P < 0.05). Conclusion the proliferation, adhesion and invasion of SKOV-3 / DDP were decreased after the inhibition of MACC1 gene expression. The increase of apoptosis may be related to the inhibition of HGF/C-met and ERK pathways.
【作者单位】：郑州市妇幼保健院;郑州大学第一附属医院;
【分类号】：R737.31

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