CTEN对雌激素受体阳性乳腺癌细胞的三苯氧胺敏感性及迁移侵袭能力的影响

Effects of CTEN on tamoxifen sensitivity,invasion and migration of estrogen receptor positive breast cancer cells

目的探讨CTEN对雌激素受体(ER)阳性乳腺癌细胞的三苯氧胺敏感性及迁移侵袭的影响。方法采用Lipofectamine 2000脂质体法分别将CTEN高表达质粒p CMV-CTEN和对照质粒p CMV-Con、CTEN干扰质粒si CTEN和对照质粒si NC及HER-2干扰质粒si HER-2和对照质粒si NC共3对质粒载体分别转染至MCF-7细胞后,采用实时定量PCR和Western blotting检测CTEN和HER-2的m RNA及蛋白水平,MTT法检测MCF-7细胞的增殖情况以评价三苯氧胺敏感性,细胞划痕实验和Transwell实验检测细胞迁移和侵袭能力。结果转染CTEN高表达质粒的MCF-7细胞中CTEN和HER-2的表达均升高,转染CTEN干扰质粒的MCF-7细胞中CTEN和HER-2的表达均降低;转染CTEN高表达质粒的MCF-7细胞对三苯氧胺的敏感性降低,细胞增殖增强,细胞迁移率升高且侵袭至下室的细胞数增多,而转染CTEN干扰质粒的MCF-7细胞则对三苯氧胺的敏感性升高,细胞增殖减弱,细胞迁移率降低且侵袭至下室的细胞数减少。在MCF-7细胞中干扰HER-2后再过表达CTEN,其促进三苯氧胺耐药及细胞增殖和迁移侵袭的作用也明显减弱。结论 CTEN具有促进ER阳性乳腺癌细胞MCF-7的三苯氧胺耐药及细胞增殖、迁移和侵袭的作用,其发生机制可能与HER-2有关。

Objective To explore the effects of CTEN on tamoxifen sensitivity, invasion and migration of estrogen receptor （ER） positive breast cancer cells. Methods Lipofectamine 2000 liposomes were used to transfect MCF-7 cells with three pairs of plasmid vectors： CTEN over-expression plasmid pCMV-CTEN and control plasmid pCMV-Con, CTEN interference plasmid siCTEN and control plasmid siNC and HER-2 interference plasmid siHER-2 and control plasmid siNC. The mRNA and protein levels of CTEN and HER-2 were detected by real-time quantitative PCR and Western blotting, respectively. The proliferation of MCF-7 cells was detected by MT/＂ as to evaluate tamoxifen sensitivity. Migration and invasive ability were detected by cell scratch test and Transwell test. Results The expression of CTEN and HER-2 in MCF-7 cells transfected with pCMV-CTEN were enhanced, while the expression of CTEN and HER-2 in MCF-7 cells transfected with siCTEN were weakened. In MCF-7 ceils transfected with pCMV-CTEN, there were reduced sen- sitivity to tamoxifen, enhanced cell proliferation and increased migration rate and number of cells invaded to the lower chamber, while opposite results were found in terms of sensitivity to tamoxifen, cell proliferation, invasion and migration ability in MCF-7 ceils trans- fected with siCTEN. The weakened tamoxifen sensitivity and enhanced cell proliferation and metastasis induced by CTEN could be re- versed by interfering HER-2. Conclusion CTEN weakened tamoxifen sensitivity and enhanced cell proliferation, migration and inva- sion ability in human ER positive breast cancer MCF-7 cells, which may act through activating HER-2.