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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 investigate the effect of chromodomain helicase (ATPD) DNA-binding protein 1-like gene (CHD1L) on invasion and migration of prostate cancer cells and its possible mechanism. Methods: The expression of CHD1L mRNA in prostate cancer cell lines LNCAP, PC3, DU145 and prostate epithelial cell line RWPE-1 was detected by real-time fluorescence quantitative PCR. The expression of CHD1L mRNA in prostate cancer PC3 cells transfected with siRNA was detected by Transwell invasion assay and Transwell invasion assay The effect of silencing CHD1L on invasion and migration of prostate cancer cells was analyzed by Western blotting. The expression of MMP-9, N-cadherin and E-cadherin in PC3 cells was detected by Western blotting. Results: The expression level of CHD1L mRNA in prostate cancer cells was significantly higher than that in prostate epithelial cells (P <0.01), and the expression of PCD3 in PC3 cells was the highest. In invasive experiments, the number of transmembrane cells in the interference group was significantly lower than that in the negative control group and the blank control group (49.67 ± 6.67 vs 113.67 ± 5.69 and 112.00 ± 12.49, P <0.05). In the scratch test, the wound healing rate at 48 h in the interference group was also lower than that in the negative control group and the blank control group [(21.27 ± 3.27)% vs (48.47 ± 5.72)% vs (49.93 ± 3.35)%, P <0.05]. The expression of MMP-9 and N-cadherin was down-regulated and the expression of E-cadherin was up-regulated in the interference group. Conclusion: Silencing CHD1L can reduce the invasion and migration of prostate cancer PC3 cells, which may be through the regulation of MMP-9 and EMT-related protein expression.