沉默CHD1L对前列腺癌PC3细胞恶性生物学行为的影响及其可能机制

Effect of CHD1L gene silencing on malignant biologic behaviors of prostate cancer PC3 cell line and its possible mechanism

目的:探讨染色质解旋酶DNA结合蛋白1样基因(chromodomain helicase/ATPase DNA binding protein 1-like gene,CHD1L)对前列腺癌细胞侵袭、迁移能力的影响及其可能的作用机制。方法:采用实时荧光定量PCR技术检测前列腺癌细胞株LNCAP、PC3、DU145以及前列腺上皮细胞株RWPE-1中CHD1L mRNA表达水平;转染siRNA干扰前列腺癌PC3细胞CHD1L的表达,并用Transwell侵袭实验和划痕实验分析沉默CHD1L对前列腺癌细胞侵袭和迁移能力的影响;Western blotting检测PC3细胞MMP-9、N-钙黏蛋白和E-钙黏蛋白的表达水平。结果:CHD1L mRNA在前列腺癌细胞中的表达水平明显高于前列腺上皮细胞(P<0.01),其中以前列腺癌PC3细胞的表达水平最高。侵袭实验中,干扰组的穿膜细胞数明显低于阴性对照组和空白对照组[(49.67±6.67)vs(113.67±5.69)和(112.00±12.49)个,P<0.05)。划痕实验中,干扰组48 h伤口愈合率也低于阴性对照组和空白对照组[(21.27±3.27)%vs(48.47±5.72)%和(49.93±3.35)%,P<0.05]。干扰组细胞MMP-9和N-钙黏蛋白表达下调,E-钙黏蛋白表达上调。结论:沉默CHD1L可降低前列腺癌PC3细胞的侵袭迁移能力,该作用可能是通过调控MMP-9和EMT相关蛋白表达实现的。

Objective：To explore the influence of CHD1L gene （chromodomain helicase/ATPase DNA binding protein 1-like gene） on the invasion and migration ability of prostate cancer cells and the potential mechanisms. Methods： Real-time fluorescent quantitative-PCR assay was carried out to detect mRNA expression of CHD1L gene in prostate cancer cell lines LNCAP, PC3 and DU 145 as well as normal prostate epithelial cell line RWPE-1. siRNA transfection was used to deplete CHD1L expression in PC3 cells. Then Transwell invasion assay and wound healing test were employed to detect the effect ofCHD1L silencing on the invasion and migration ability of PC3 cells. Protein expressions of MMP-9, N-cadherin and E-cadherin were examined by Western blotting assay. Results： Compared to normal prostate epithelial cells, three prostate cancer cell lines all showed significantly higher expression of CHD1L mRNA （P〈0.01）, and PC3 cells demonstrated the highest CHD1LmRNA expression. Invasion assay showed that the trans-membrane cell number in CHD1L silencing group was significantly lower than that in negative and blank control groups （49.67±6.67vs 113.67±5.69 and 112.00±12.49, P〈0.05）. Wound healing test also exhibited lower healing degree in CHD1L silencing group than that in negative and blank control groups at 48 h （[21.27±3.27]%vs [48.47±5.72]% and [49.93±3.35]%, respectively, P〈0.05）. Furthermore, protein expressions of MMP-9 and N-cadherin decreased while E-cadherin increased in CHD1L gene depletion cells. Conclusions： CHD1L silencing could dampen the invasion and migration ability of prostate cancer PC3 cell line, and this might be accomplished by regulating MMP-9 and EMT related proteins.