| 冯 茜,周 汝,余沈桐,杨 桐,于舒鸿,刘 爽,高继伟,崔竹青,张 静.血管细胞黏附分子-1对卵巢癌细胞凋亡的影响及机制研究[J].,2018,(9):1658-1663 |
| 血管细胞黏附分子-1对卵巢癌细胞凋亡的影响及机制研究 |
| Effect and Mechanism of Vascular Cell Adhesion Molecule-1 on Cell Apoptosis of Ovarian Carcinoma |
| 投稿时间：2017-11-28 修订日期：2017-12-23 |
| DOI：10.13241/j.cnki.pmb.2018.09.012 |
| 中文关键词: 血管细胞黏附分子-1 卵巢癌 细胞凋亡 STAT3 |
| 英文关键词: Vascular cell adhesion molecule-1 Ovarian carcinoma Apoptosis STAT3 |
| 基金项目:国家自然科学基金项目（81372783） |
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| 中文摘要: |
| 摘要 目的：探讨过表达血管细胞黏附分子-1（vascular cell adhesion molecule-1，VCAM-1）对卵巢癌细胞凋亡的影响。方法：构建过表达VCAM-1的慢病毒载体GV358-VCAM1+，转染人类卵巢癌IGROV1细胞株，利用嘌呤霉素筛选稳定表达VCAM-1的IGROV1细胞，通过倒置荧光显微镜下观察绿色荧光，确定细胞转染效率，Western blot及RT-PCR法确定卵巢癌细胞VCAM-1蛋白和mRNA水平；采用流式细胞仪检测过表达VCAM-1的IGROV1的细胞凋亡变化，western blot法检测凋亡相关蛋白（Bcl-2、Bax、Casepase-3、Cleaved Casepase-3）以及STAT3、p-STAT3蛋白表达水平的变化。结果：成功构建的慢病毒载体GV358-VCAM1+在IGROV1细胞中的转染效率达到85%以上，转染细胞的VCAM-1蛋白及mRNA水平均呈稳定表达；VCAM-1过表达卵巢癌细胞的细胞凋亡显著高于空载体对照组（P=0.0149）；Bax、Casepase-3、Cleaved Casepase-3表达水平均较对照组显著升高（P<0.01），Bcl-2、p-STAT3表达水平明显低于对照组（P<0.01），但STAT3表达水平无显著改变。结论：VCAM-1可能通过下调STAT3的磷酸化水平诱导卵巢癌细胞凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of the overexpression of vascular cell adhesion molecule-1 on the apoptosis of ovarian carcinoma cells. Methods: The lentiviral vector with full length VCAM-1 gene (GV358-VCAM1+) was constructed and trans- fected into human ovarian carcinoma cell line, IGROV1. The tumor cells with VCAM-1 stable overexpression were selected by puromycin. The transfection rate was determined via the observation of green fluorescence under an inverted fluorescence microscope. The levels of VCAM-1 protein and mRNA were identified by western blot and RT-PCR analysis, respectively. The cell apoptosis was an- alyzed by flow cytometry assay. Meanwhile, the expression level of apoptosis related proteins (Bcl-2, Bax, Casepase-3 and Cleaved Casepase-3), STAT3 and p-STAT3 protein, were detected by western blot analysis. Results: The transfection rate of GV358-VCAM1+ in IGROV1 cells was more than 85%. These cells had stable VCAM-1 expression not only at the protein but also mRNA level. Tumor cells with VCAM-1 overexpression had significantly higher apoptosis than those with empty vector control group (P=0.0149). Moreover, compared with the empty vector control group, there were significantly higher expression of Bax, Casepase-3 and Cleaved Casepase-3(P<0.01), and lower expression of Bcl-2 and p-STAT3 in VCAM-1 overexpression group(P<0.01). However, the expression level of STAT3 was not significantly changed. Conclusion: VCAM-1 could induce the cell apoptosis of ovarian carcinoma through down-regula- tion of STAT3 phosphorylation. |
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