核因子E2相关因子2基因沉默对人前列腺癌PC-3细胞增殖、侵袭和转移的影响

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Abstract Objective To investigate the effect of NRF2 gene silencing on proliferation, invasion and metastasis of human prostate cancer PC-3 cells and the mechanism.Methods The expressions ofNRF2 mRNA in human prostate epithelial cell line RWPE-1 and prostate cancer cell line PC-3 were detected by RT-PCR.After the designed NRF2 siRNA-1, NRF2 siRNA-2 andNRF2 siRNA-3 interference sequences were transfected into PC-3 cells by lipofection, the bestNRF2 siRNA sequence was screened by Western blot and RT-PCR. PC-3 cells were divided into Con group (untransfected), NC group (transfected negative control) and NRF2siRNA group (transfected NRF2 siRNA-3). The proliferation, cloning, invasion and migration of each group were detected with CCK-8 method, plate cloning test and Transwell test,while the expressions of Ki67,CyclinD1, MMP-9 and N-cadherin proteins in each group were detected by Western blot.Results Compared with prostatic epithelial cell line RWPE-1, the expression of NRF2mRNA in prostate cancer cell line PC-3 was significantly increased (0.84±0.05 vs 0.22±0.03,P<0.05). The expressions of NRF2 protein(0.56±0.04,0.39±0.03,0.11±0.02 vs 0.76±0.07) and mRNA(0.80±0.05,0.52±0.03,0.26±0.03 vs 1.00±0.05) in PC-3 cells could be down-regulated by NRF2 siRNA-1, NRF2 siRNA-2 and NRF2 siRNA-3 interfering sequences,and the interference effect of NRF2 siRNA-3 was significantly greater than that of NRF2 siRNA-1 andNRF2 siRNA-2.Compared with Con group, the proliferation, cloning, invasion and migration abilities[survival rate:24 h (82.05±5.24)% vs (99.86±5.05)%,48 h (64.57±4.85)% vs(97.28±6.36)%,72 h (45.62±3.76)% vs(94.57±5.49)%;clone formation rate:(39.35±3.26)% vs(66.52±4.85)%;number of invasive cells:42.00±5.50 vs 85.00±7.50;number of migrating cells:58.00±3.00 vs 118.50±8.00]of NRF2 siRNA group were significantly weakened, while the expressions of Ki67, Cyclin D1, MMP-9 and N-cadherin (Ki67:0.25±0.03 vs 0.78±0.05;CyclinD1 0.41±0.03 vs 0.85±0.06;MMP-9:0.10±0.02 vs 0.89±0.07; N-cadherin:0.22±0.02 vs 0.49±0.04)were significantly decreased (P<0.05), but there was no significant difference between NC group and Con group (P>0.05).Conclusions NRF2gene silencing can inhibit the proliferation, invasion and metastasis of human prostate cancer PC-3 cells, and the molecular mechanism may be related to down-regulation of the expressions of Ki67, Cyclin D1, MMP-9 and N-cadherin proteins.

Key words： prostate cancer NRF2 gene cell proliferation cell invasion cell migration

Received: 15 March 2019

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