ABSTRACT
Objective: To investigate the effects of WNT5B over-expression on the invasion and migration of human breast cancer MDA-MB-231 cells, and to explore its possible mechanisms. Methods: After the recombinant vector pcDNA3.1-WNT5B+EGFP containing enhanced green fluorescent protein (EGFP) and WNT 5B genes was transfected into human breast cancer MDA-MB-231 cells, the expression levels of WNT5B mRNA and protein were examined by real-time fluorescent quantitative PCR and Western blotting, respectively. The abilities of cell migration and invasion were analyzed by wound-healing test and Transwell chamber assay, respectively. Furthermore, the expression levels of epithelial-mesenchymal transition (EMT)-related molecules N-cadherin, E-cadherin and Vimentin were detected by Western blotting. Results: Compared with the untransfection and the empty vector-transfection groups, the expression levels of WNT5B mRNA (P < 0.01) and protein (P < 0.05) in MDA-MB-231 cells transfected with the recombinant vector pcDNA3.1-WNT5B+EGFP were up-regulated. The abilities of migration (P < 0.05) and invasion (P < 0.01) of MDA-MB-231 cells after WNT5B over-expression were significantly decreased. The expression level of E-cadherin in MDAMB- 231 cells with WNT5B over-expression was up-regulated (P < 0.05), whereas the expression levels of N-cadherin and Vimentin were down-regulated (both P < 0.05). Conclusion: WNT5B over-expression can inhibit the migration and invasion of breast cancer MDA-MB-231 cells, which may be related to the regulation of EMT process.
ABSTRACT
AIM: To detect the expression of WNT5B in normal breast epithelial cells and different breast cancer cell lines, and to investigate the effects of WNT5B over-expression on the viability and apoptosis of human breast cell line MCF-7.METHODS: The mRNA expression of WNT5B was detected by RT-PCR in different breast cancer cells. MCF-7 cells were transfected with plasmid pcDNA3.1 /WNT5B or pcDNA3.1, and the expression of WNT5B at mRNA and protein levels was examined in the 2 groups by real-time PCR and Western blotting, respectively.Subsequently, the chan-ges of cell viability and cell apoptosis were analyzed by CCK-8 assay and flow cytometry, respectively.RESULTS: The ex-pression of WNT5B in the breast cancer cell lines was lower than that in MCF10A cells.The WNT5B expression in the MCF-7 cells in experimental group was significantly higher than that in vector group (P <0.05).However, the cell viabili-ty in experimental group decreased significantly as compared with vector group (P <0.05).The number of the cells in S-phase obviously increased, while the percentage of the cells in G1-phase and G2 /M-phase decreased compared with vector group.The number of apoptotic cells in WNT5B group was significantly higher than that in vector group.CONCLUSION:The expression of WNT5B is decreased in breast cancer cells.WNT5B over-expression significantly inhibits the cell growth and promotes the cell apoptosis in breast cancer MCF7 cells.
ABSTRACT
[ ABSTRACT] AIM:To study the expression of WNT5B in the breast cancer and further to discuss the correlation between WNT5B and clinicopathologic characteristics of breast cancer.METHODS:The expression of WNT5B at mRNA and protein levels was measured by real-time PCR and Western blot in 67 cases of breast cancer and the tissue adjacent to carcinoma.In addition, the immunohistochemical method was used to detect the expression of WNT5B in the breast cancer and the tissue adjacent to carcinoma.The relationships between WNT5B expression and clinicopathologic indexes were also analyzed.RESULTS:The expression of WNT5B in the breast cancer was obviously lower than that in the tissue adjacent to carcinoma (P20 mm group (P0.05) in the breast cancer. CONCLUSION:The expression of WNT5B decreases obviously in breast cancer.The expression of WNT5B is related to primary tumor size, which provides new ideas for the diagnosis and treatment of breast cancer, suggesting that WNT5B may be a new molecular marker for prognosis of breast cancer.