Effects of transfection of tbx2 siRNA on cell proliferation, migration, invasion, and EMT of human prostate cancer cells

FANG Qian; DU Wenliang; WANG Junqi; TENG Jingwei; CHEN Yue; XIAO Yongshuang

doi:10.3969/j.issn.1002-266X.2017.23.010

Abstract：Objective To observe the effects of transfection of tbx2 siRNA on cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of human non-androgen dependent prostate cancer cells PC3 and androgen-dependent prostate cancer cells LNCaP.Methods Human prostate cancer PC3 and LNCaP cells cultured in vitro were randomly divided into the observation group and the control group, which were transfected with tbx2 siRNA and negative control siRNA by SiLentFectTM Lipid Reagent, respectively, and then were cultured for 48 h.The protein expression of tbx2 in cells of the two groups was detected by Western blotting.The proliferative activity (OD450) was detected by using cell counting Kit (CCK-8).Transwell migration and invasion assay were performed to detect the migration and invasion cells of both groups.The expression of EMT-related markers including E-cadherin, N-cadherin, Vimentin, and Fibronectin, and EMT-related upstream transcription factors Snail and Twist, was detected by Western blotting.We did the same experiments for LNCaP cells.Results Tbx2 siRNA was successfully transfected into two kinds of prostate cancer cells.The expression levels of tbx2 cultured for 48 h in PC3 cells of the observation group and control group were 0.345±0.016 and 0.723±0.31, respectively, and they were 0.315±0.006 and 0.92±0.10, respectively, in LNCaP cells.The relative expression of tbx2 in PC3 and LNCaP cells of the observation group was lower than that in the control group (both P<0.05).After being cultured for 24, 48, and 72 h, the OD450 of PC3 was 0.487 0.026, 0.868 0.031, and 1.387±0.107, respectively, in the control group, versus 0.315±0.022, 0.550±0.027, and 0.765±0.046 in the observation group.The OD450 of LNCaP was 0.388±0.032, 0.722±0.032, and 1.187 0.135, respectively, in the control group, versus 0.251±0.021, 0.450±0.037, and 0.625±0.066 in the observation group.The OD450 of PC3 and LNCaP cells in the observation group was lower than that in the control group, respectively (both P<0.05).The number of migration cells in PC3 cells was 118.67±6.3 and 292.33±5.03, versus 85.67 4.31 and 192.33 8.07 in LNCaP cells, respectively, in the observation group and the control group.The number of migration cells of PC3 and LNCaP in the observation group was lower than that in the control group, respectively (both P<0.05).The number of invasion cells in PC3 cells was 89.00±3.02 and 186.33±5.84, versus 81.67±3.42 and 232.33±742 in LNCaP cells, respectively.The number of invasion cells of PC3 and LNCaP in the observation group was lower than that in the control group, respectively (both P<0.05).The expression of N-cadherin, Vimentin, Fibronectin, Snail, and Twist in PC3 and LNCaP cells of the observation group was significantly lower, while the expression of E-cadherin protein was higher than that in the control group (all P<0.05).Conclusion Silencing tbx2 gene can inhibit the proliferation, decrease the abilities of migration and invasion, and inhibit EMT process in prostate cancer cells.