Yun Fu1, Bei-qi Jiang1, Yi Wu1, Zheng-dong Li1, Zhi-gang Zhuang2. 1. Department of Breast Surgery, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, China. 2. Department of Breast Surgery, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, China. Email:zhuang_zg@163.com.
Abstract
OBJECTIVE: To explore the effects of hsa-miR-206 on the proliferation, migration and invasion of breast cancer. METHODS: The hsa-miR-206 mimics, inhibitors and their paired negative controls were transfected into human breast cancer cell line MDA-MB-231 by liposome. The proliferation of cell was evaluated by CCK-8 and the migration and invasion was detected by Transwell. Matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9), breast cancer metastasis-suppressor 1 (BRMS-1) and connexin 43 (Cx43) were detected by both quantitative polymerase chain reaction (qPCR) and Western blot. The expression of miR-206 was detected by qPCR. Dual luciferase assay was detected to confirm the specific binding sites of miR-206 and Cx43. RESULTS: (1) The proliferation activity of 206m-group cell (0.74 ± 0.16) was significantly lower than that of control group (1.12 ± 0.23) (t = -3.066, P = 0.037) while that of 206i-group cell (1.43 ± 0.26) was higher than that of control group (0.98 ± 0.14) (t = 3.635, P = 0.022). (2) Transwell tests showed the migration and invasion of 206m-group cell decreased significantly (migration:0.56 ± 0.01 vs 0.63 ± 0.01, t = -23.00, P = 0.002; invasion:0.79 ± 0.01 vs 0.99 ± 0.01, t = -21.200, P = 0.002), but that of 206i-group cell increased significantly (migration:0.97 ± 0.11 vs 0.61 ± 0.09, t = 32.787, P = 0.001; invasion:1.10 ± 0.01 vs 0.93 ± 0.05, t = 5.167, P = 0.035). (3) The expressions of MMP-2,MMP-9 and Cx43 decreased and the expression of BRMS-1 increased in 206m-group cell and vice versa in 206i group. (4) The expression of miR-206 in lymph node-negative group of clinical breast cancer sample was higher than that of lymph node-positive one. And there was statistical difference (Z = -2.098, P = 0.003). And the expression of Cx43 was opposite. (5) Dual luciferase reporter assay confirmed the specific binding sites of hsa-miR-206 and Cx43. CONCLUSION: Hsa-miR-206 has negative controls of proliferation, migration and invasion of breast cancer cell by targeting Cx43.
OBJECTIVE: To explore the effects of hsa-miR-206 on the proliferation, migration and invasion of breast cancer. METHODS: The hsa-miR-206 mimics, inhibitors and their paired negative controls were transfected into humanbreast cancer cell line MDA-MB-231 by liposome. The proliferation of cell was evaluated by CCK-8 and the migration and invasion was detected by Transwell. Matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9), breast cancer metastasis-suppressor 1 (BRMS-1) and connexin 43 (Cx43) were detected by both quantitative polymerase chain reaction (qPCR) and Western blot. The expression of miR-206 was detected by qPCR. Dual luciferase assay was detected to confirm the specific binding sites of miR-206 and Cx43. RESULTS: (1) The proliferation activity of 206m-group cell (0.74 ± 0.16) was significantly lower than that of control group (1.12 ± 0.23) (t = -3.066, P = 0.037) while that of 206i-group cell (1.43 ± 0.26) was higher than that of control group (0.98 ± 0.14) (t = 3.635, P = 0.022). (2) Transwell tests showed the migration and invasion of 206m-group cell decreased significantly (migration:0.56 ± 0.01 vs 0.63 ± 0.01, t = -23.00, P = 0.002; invasion:0.79 ± 0.01 vs 0.99 ± 0.01, t = -21.200, P = 0.002), but that of 206i-group cell increased significantly (migration:0.97 ± 0.11 vs 0.61 ± 0.09, t = 32.787, P = 0.001; invasion:1.10 ± 0.01 vs 0.93 ± 0.05, t = 5.167, P = 0.035). (3) The expressions of MMP-2,MMP-9 and Cx43 decreased and the expression of BRMS-1 increased in 206m-group cell and vice versa in 206i group. (4) The expression of miR-206 in lymph node-negative group of clinical breast cancer sample was higher than that of lymph node-positive one. And there was statistical difference (Z = -2.098, P = 0.003). And the expression of Cx43 was opposite. (5) Dual luciferase reporter assay confirmed the specific binding sites of hsa-miR-206 and Cx43. CONCLUSION: Hsa-miR-206 has negative controls of proliferation, migration and invasion of breast cancer cell by targeting Cx43.
Authors: Eleni van Schooneveld; Hans Wildiers; Ignace Vergote; Peter B Vermeulen; Luc Y Dirix; Steven J Van Laere Journal: Breast Cancer Res Date: 2015-02-18 Impact factor: 6.466
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