Integrin β3 down-regulates invasive features of ovarian cancer cells in SKOV3 cell subclones
Integrin β3 down-regulates invasive features of ovarian cancer cells in SKOV3 cell subclones
复制标题
DOI：
10.1007/s00432-008-0526-8
复制
发表时间：
2009-07-01
影响因子：
3.6
通讯作者：
Fu, Maosun
中科院分区：
文献类型：
--
作者：
Chen, Jie;Zhang, Jie;Fu, Maosun
为了探讨整合素β 3在卵巢癌SKOV 3细胞侵袭性中的作用，通过比较不同的转移亚克隆，本研究采用有限稀释法从人卵巢癌亲代细胞系SKOV 3中分离并建立了两个分别具有高转移潜力和低转移潜力的细胞亚克隆，命名为S1和S21。采用实时荧光定量RT-PCR和流式细胞仪检测两种细胞亚克隆中整合素α v、整合素α v β 3、整合素β 3、E-钙粘素、FAK和ILK的表达。随后，将整合素β 3的siRNA转染S21细胞，通过MTT法、集落形成实验、Matrigel侵袭实验和细胞迁移实验检测转染后S21细胞中整合素α v β 3、整合素β 3和E-cadherin的表达明显下调;而整合素α v、FAK和ILK β的表达无显著差异。值得注意的是，通过siRNA技术获得了超过70%的整联蛋白β 3表达的敲低。整合素β 3-siRNA转染的细胞显示出与模拟转染的细胞相比，细胞增殖、细胞迁移和侵袭活性显著增加。整合素β 3-siRNA转染的SKOV 3细胞中整合素α v β 3和E-cadherin的表达均低于对照组，整合素β 3可能与E-cadherin一样，是卵巢癌细胞侵袭性的抑制基因。
To investigate the role of integrin beta 3 in invasive features of ovarian cancer SKOV3 cells, by comparing different metastatic subclones.In the present study, two cell subclones, termed as S1 and S21, which possessed high and low metastatic potential, respectively, were isolated and established from human ovarian cancer parental cell line SKOV3 by the limited dilution method. The expressions of integrin alpha v, integrin alpha v beta 3, integrin beta 3, E-cadherin, FAK and ILK in the two cell subclones were compared by means of real-time RT-PCR or flow cytometry. Subsequently, S21 was transfected with siRNA for integrin beta 3 and the effects of transfection were examined by methyl thiazolyl tetrazolium (MTT) assay, colony formation assay, Matrigel invasion assay and cell migration assay.The expressions of integrin alpha v beta 3, integrin beta 3 and E-cadherin were markedly down-regulated in S1; however, there were no significant differences in the expressions of integrin alpha v, FAK and ILK beta. Of note, more than 70% knockdown of integrin beta 3 expression was obtained by siRNA technique. The integrin beta 3-siRNA-transfected cells showed significant increases in cell proliferation, cell migration and invasive activity in contrast with the mock-transfected cells. The expressions of integrin alpha v beta 3 and E-cadherin were lower in the integrin beta 3-siRNA-transfected cells compared to the mock control.Integrin beta 3, like E-cadherin, may be also a suppressor gene down-regulating invasive features of ovarian cancer cells in SKOV3 cell subclones.