Influence Of Silencing Med19on Metastasis And Invasion Of Human Breast Cancer MCF-7Cells

Objective To study the influence of silencing Med19on metastasis and invasion ofhuman breast cancer MCF-7cells， further explored Med19gene role and activemechanism in metastasis and invasion of the breast cancer.Method Construction of lentivirus expression vector according to the sequence ofMed19gene, then transfection of human breast cancer MCF-7cell line withsiRNA-Med19lentirivus expression vector. RT-PCR and Western blot were used todetermine transcription levels of Med19gene mRNA and protein expression, scratchingand transwell tests were applied to determine the ability of metastasis and invasion ofcells. RT-PCR was used to determine the expressions of MMP-9and VEGF mRNA.Results siRNA-Med19lentirivus expression vector was transfected into humanbreast cancer MCF-7cells successfully. The Med19mRNA expressions of the group ofMCF-7cells which transfected siRNA-Med19lentivirus expression vector(KD group),negative control group(NC group) and control group(CON group) were0.076±0.033,0.274±0.010,0.272±0.057. Compared with MCF-7cells of negative controlgroup and control group, the Med19expression of KD group decreased by72.3%and72.1%and the differences were significant(P<0.05). The Med19protein expressions of KDgroup, NC group and CON group were0.106±0.037,0.725±0.134and0.721±0.135. TheMed19protein expression of KD group decreased by85.4%and85.3%compared to NCgroup and CON group. The level of Med19protein expression of KD group decreasedsignificantly(P<0.05). The cells passed through polycarbonate membrane of KD group,NC group and CON group were23.8±4.32,43.4±3.65and45.8±5.81. The metastatic abilityof KD group was significant reduced(P<0.05). The invasive distances of KD group, NC group and CON group MCF-7cells were compared at the time of6hour,12hour and24hour. The invasive ability of MCF-7cells decreased significantly with the extension oftime(P<0.05). The MMP-9mRNA expressions of KD group, NC group and CON groupwere361±42.899,581±42.723and588±18.330. The MMP-9mRNA expression of KDgroup decreased by37.87%and38.61%and the differences were significant (P<0.05).TheVEGF mRNA expressions of KD group, NC group and CON group were1307.667±75.295,2067.667±54.123and2096±151.615. The VEGF mRNA expression ofKD group decreased by36.77%and37.64%and the differences were significant(P<0.05).The expressions of Med19mRNA and protein of NC group had no significant differencefrom CON group(P>0.05). The differences of the ability of cells metastasis and invasionwere not significant (P>0.05). The differences of the expressions of MMP-9and VEGFmRNA were not significant(P>0.05).Conclusion1、 Construction of siRNA-Med19lentirivus expression vectortransfected into human breast cancer MCF-7cells can decrease the expressions of Med19mRNA and protein of human breast cancer MCF-7cells.2、Silencing of Med19gene can decrease human breast cancerMCF-7cells ability of metastasis and invasion,then suppress the malignant biologicalbehavior of breast cancer.3、Silencing of Med19gene can decrease the expressions of MMP-9and VEGF mRNA, so suppress the metastasis and invasion of breast cancer cells.