Influence of silencing Med19 on metastasis and invasion of human breast cancer MCF-7 cells
10.3969/j.issn.1006-5725.2014.09.010
- VernacularTitle:沉默中介体19对人乳腺癌MCF-7细胞侵袭及迁移的影响
- Author:
Xiaoyan WANG
;
Zijian GUO
;
Xiaohong WU
;
Lihua LI
- Publication Type:Journal Article
- Keywords:
Med19;
Lentivirus;
Breast cancer cells;
RNA interference
- From:
The Journal of Practical Medicine
2014;(9):1373-1375
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the influence of silencing Med19 on metastasis and invasion of human breast cancer MCF-7 cells. Methods lentivirus expression vector delivering small hairpin RNA (shRNA) against Med19 gene was constructed, then transfected of human breast cancer MCF-7 cell line. There were three experimental groups: non-infected (CON) group, Lv-NC-infected (NC) group, and Lv-shMed19-infected (KD) group. To determine the lentiviral infection efficiency, expression of GFP was detected with fluorescence microscopy. The mRNA and protein levels of Med19 in three groups MCF-7 cells were detected by real-time PCR and Western blotting. Scratching and transwell tests were employed to determine the ability of metastasis and invasion of cells. Results The highest infection efficiency was obtained, resulting fluorescent expression identified in more than 90%of MCF-7 cells 120 h after infection. The expression of Med19 mRNA in the KD group was dramatically decreased by 72.3%compared with the NC group, and by 72.1% compared with the CON group, respectively (P < 0.05). The Med19 protein expression in the KD group was significantly lower than that of the NC group and the CON group, with a reduction of 85.4%and 85.3%, respectively (P<0.05). No statistical significance was detected between the NC groups and the CON groups. Thus, the constructed Lv-shMed19 was demonstrated to be active and specific in inhibiting the expression of Med19. Moreover, the invasive distances of KD group, NC group and CON group MCF-7 cells were compared at 6 hour, 12 hour and 24 hour. The invasive ability of MCF-7 cells decreased significantly with the extension of time (P<0.05). The cells passed through polycarbonate membrane in KD group, NC group and CON group were 23.8 ± 4.32, 43.4 ± 3.65 and 45.8 ± 5.81 respectively. The metastatic ability of KD group was significant reduced (P<0.05). Conclusion Silencing of Med19 gene significantly decreases the ability of metastasis and invasion of human breast cancer MCF-7 cells, and then suppresses the malignant biological behavior of breast cancer.
