4-OHT inhibits cell migration by reversal of eMT in triple negative breast cancer cells / 中国医师进修杂志

ABSTRACT

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.