4-羟基他莫昔芬逆转三阴性乳腺癌细胞间质表型降低侵袭转移能力的机制研究  

4-OHT inhibits cell migration by reversal of eMT in triple negative breast cancer cells

作　　者：汪茜[1] 王一丁[2] Wang Qian;Wang Yiding(Department of Oncology, Liaoning Cancer Hospital & Cancer Hospital of China Medical University,Shenyang 110042, China;Department of Urology Surgery, Liaoning Cancer Hospital & Cancer Hospitalof China Medical University, Shenyang 110042, China)

机构地区：[1]辽宁省肿瘤医院中国医科大学附属肿瘤医院肿瘤内科,沈阳110042 [2]辽宁省肿瘤医院中国医科大学附属肿瘤医院泌尿外科,沈阳110042

出　　处：《中国医师进修杂志》2019年第6期485-491,共7页Chinese Journal of Postgraduates of Medicine

基　　金：2017年辽宁省博士科研启动基金指导计划项目(20170520005).

摘　　要：目的探讨4-羟基他莫昔芬(4-OHT)能否通过逆转三阴性乳腺癌(TNBC)细胞间质表型降低细胞侵袭转移的能力。方法采用倒置显微镜观察细胞形态、Transwell法测定细胞迁移能力、Western blot检测E-cadherin,Vimentin和Actin蛋白的表达、免疫荧光显微技术观察E-cadherin,Vimentin蛋白的分布、Real-time聚合酶链式反应(PCR)技术检测微小RNA(miR)-200c表达观察细胞侵袭转移能力的变化。动物实验建立乳腺癌肺转移模型,检测裸鼠肺组织标本中乳腺癌细胞的侵袭转移情况。结果4-OHT能够通过上调TNBC中E-cadherin表达,下调Vimentin表达逆转上皮间质转化(EMT);Transwell实验证实在MDA-MB-231和MCF-7/ADR细胞中4-OHT能够分别降低细胞的迁移能力至59.4%和43.2%;4-OHT处理后细胞转移能力减退,肺转移成瘤率降至66.7%;4-OHT能够将TNBC MDA-MB-231和MCF-7/ADR中miR-200c的相对表达量分别升高至280%和450%,而MCF-7的miR-200c水平无明显变化;抑制miR-200c的表达后,4-OHT不能够降低TNBC细胞的侵袭迁移能力。结论4-OHT能够上调miR-200c的表达,逆转三阴性乳腺癌细胞间质表型降低细胞侵袭转移的能力。Objective To investigate whether 4-OHT could decrease the ability of invasion and metastasis by reversal of EMT in TNBC. Methods Inverted microscope was used for observing cell morphological change. Cell migration ability was measured using transwell assay. Expression of E-cadherin, Vimentin and Actin proteins were analyzed by Western blot. The distribution of E-cadherin and Vimentin were analyzed by immune- fluorescence microscopy. Relative levels of miR-200c were measured by Real-time PCR. Transient transfection was performed using Lipofectamine 2000 reagent. Animal model was established to test the migration rate of lung metastasis and metastase focals. The HE staining was performed in mice lung tissue using Immunohistochemistry to confirm the metastasis. Results 4-OHT upregulates E-cadherin expression, downregulates vimentin expression to reverse EMT in TNBC cells. 4-OHT decreases migration ability of MDA-MB-231 and MCF-7/ADR cells by transwell test to 59.4% and 43.2%;The migration ability of cells decrease to 66.7%. 4-OHT up-regulates the expression of miR-200c in MDA-MB-231 and MCF-7/ADR cells to 280% and 450%. Inhibits expression of miR-200c, 4-OHT cannot decrease migration ability of TNBC cells. Conclusions 4-OHT could decrease the ability of invasion and metastasis, up-regulate miR-200c and reverse EMT in TNBC.

关 键 词：乳腺肿瘤 肿瘤侵润 4-轻化他莫昔芬 上皮间质转化 

分 类 号：R737.9[医药卫生—肿瘤]