整合素α5表达上调促进卵巢癌SKOV3细胞侵袭转移及机制探讨

Promoting effect of upregulation of integrin α5 on invasion and metastasis of ovarian cancer SKOV3 cells and discussion on mechanism

目的:检测整合素α5(integrinα5,ITGA5)表达上调对卵巢癌SKOV3细胞侵袭转移影响及机制探讨。方法:用pEGFP-ITGA5过表达质粒对卵巢癌SKOV3细胞进行转染,未转染的SKOV3细胞作为对照。RT-PCR方法检测SKOV3细胞中整合素α5的mRNA水平变化,Western Blot方法检测整合素α5蛋白水平的表达。为观察整合素α5表达上调对SKOV3细胞侵袭转移能力的影响,采用Transwell小室检测细胞侵袭及迁移能力。小鼠体内粘附实验观察整合素α5过表达的SKOV3-GFP细胞体内粘附能力变化。Western Blot方法检测EMT相关蛋白EMT相关基因表达情况。结果:与对照组相比,pEGFP-ITGA5转染SKOV3细胞后整合素α5基因表达水平明显增高。细胞侵袭、迁移实验表明,pEGFP-ITGA5转染组较对照组细胞侵袭、迁移力明显增加,对照组和实验组穿膜细胞数分别为(56.32±4.63)个和(135.58±7.94)个,对照组与实验组相比差异有统计学意义(P<0.05)。小鼠体内粘附实验证明,整合素α5过表达的SKOV3-GFP细胞体内粘附能力明显增强。Western Blot结果表明整合素α5过表达的SKOV3中,E-cadherin表达明显下调,同时Vimentin、MMP-9表达明显上调。结论:整合素α5表达上调能促进卵巢癌SKOV3细胞的粘附和侵袭转移,其可能是通过促进上皮间质转化发生进而引起细胞侵袭转移能力的增强。

Objective： To detect the promoting effect of upregulation of integrin α5 （ ITGA5 ） on invasion and metastasis of ovarian cancer SKOV3 cells and explore the mechanism. Methods： pEGFP - ITGA5 over - expression plasmid was used to transfect ovarian cancer SKOV3 cells （ experimental group） , the ovarian cancer SKOV3 cells without transfection were selected as control group. The change of IT- GA5 mRNA expression in ovarian cancer SKOV3 cells was detected by RT - PCR, the expression of ITGA5 protein was detected by Western Blot. Transwell was used to detect the invasion and migration of ovarian cancer SKOV3 cells. In vivo adhesion assay was used to observe the change of in vitro adhesion ability in ovarian cancer SKOV3 cells with ITGA5 over - expression. Western Blot was used to detect the expression of epithelial - mesenchymal transition （EMT） - related proteins and genes. Results： Compared with control group, the expression level of ITGA5 gene in experimental group increased significantly. Invasion and metastasis increased in experimental group compared with control group, the numbers of cell permeating septum in control group and experimental group were （56. 32 ± 4. 63 ） and （ 135.58 ±7. 94）, respectively, there was statistically significant difference between the two groups （ P 〈 0. 05 ） . In vitro adhesion ability strengthened significantly in ovarian cancer SKOV3 cells with ITGA5 over - expression. In ovarian cancer SKOV3 ceils with ITGA5 over - expression, E - cadherin expression was down - regulated, while Vimentin and MMP - 9 expressions were up - regulated. Conclusion： Up - regulation of IT- GAS can promote invasion and metastasis of ovarian cancer SKOV3 cells, which may take effect by promoting EMT.