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目的:探讨下调Krüppel样因子4(Krüppel-like factor 4,KLF 4)对前列腺癌细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)和细胞迁移及侵袭能力的影响。方法:构建稳定敲降KLF 4的前列腺癌LNCaP-shKLF4细胞和对照组LNCaP-con细胞。应用实时荧光定量PCR及蛋白质印迹法检测LNCaPshKLF4和LNCaP-con细胞中KLF4 mRNA和蛋白的表达,以及稳定敲降KLF 4的前列腺癌PC3-shKLF4细胞、对照组PC3-con细胞、LNCaPcon细胞和LNCaP-shKLF4细胞中EMT相关分子mRNA和蛋白的表达。Transwell小室法检测PC3-con、PC3-shKLF4、LNCaP-con和LNCaPshKLF4细胞的迁移及侵袭能力。结果:成功构建LNCaP-shKLF4和对照组LNCaP-con细胞。LNCaPshKLF4细胞中KLF4 mRNA及蛋白表达水平均低于对照组LNCaP-con细胞(P<0.01,P<0.05)。PC3-shKLF4和LNCaP-shKLF4细胞中E-钙黏蛋白(E-cadherin,E-cad)mRNA的表达水平分别高于对照组的PC3-con和LNCaP-con细胞(P值均<0.01),而PC3-shKLF4和LNCaPshKLF4细胞中N-钙黏蛋白(N-cadherin,N-cad)、锌指E-盒结合同源异体框1(Zinc finger E-box-binding homeobox 1,Zeb1)、Snail1、波形蛋白(vimentin,Vim)和基质金属蛋白酶1(matrix metallopeptidase 1,MMP1)mRNA的表达水平均分别低于对照组的PC3-con和LNCaP-con细胞(P值均<0.05)。PC3-shKLF4和LNCaP-shKLF4细胞中E-cad的表达水平分别高于对照组的PC3-con和LNCaP-con细胞(P值均<0.05),而PC3-shKLF4和LNCaP-shKLF4细胞中N-cad、Zeb1、Snail1、Vim和MMP1蛋白的表达水平均分别低于对照组的PC3-con和LNCaP-con细胞(P值均<0.05)。PC3-shKLF4和LNCaP-shKLF4细胞的迁移和侵袭能力均分别低于对照组PC3-con和LNCaP-con细胞(P<0.01,P<0.05)。结论:下调前列腺癌细胞中KLF4表达可促进上皮相关基因的表达,抑制间质相关基因的表达,从而在体外抑制前列腺癌细胞的迁移和侵袭。
Objective: To investigate the effects of down-regulation of Krüppel-like factor 4 (KLF 4) on epithelial-mesenchymal transition (EMT) and cell migration and invasion in prostate cancer cells. Methods: To construct LNCaP-shKLF4 prostate cancer cells stably knocked down KLF 4 and control group LNCaP-con cells. The expression of KLF4 mRNA and protein in LNCaPshKLF4 and LNCaP-con cells was detected by real-time fluorescence quantitative PCR and Western blotting, and PC3-shKLF4 cells in prostate cancer cells, LNCaPcon cells and LNCaP- Expression of EMT related molecules mRNA and protein in shKLF4 cells. Transwell chamber assay was used to detect the migration and invasion of PC3-con, PC3-shKLF4, LNCaP-con and LNCaPshKLF4 cells. Results: LNCaP-con cells were successfully constructed in LNCaP-shKLF4 and control groups. The expression of KLF4 mRNA and protein in LNCaPshKLF4 cells was lower than that in LNCaP-con cells (P <0.01, P <0.05). The expression of E-cadherin mRNA in PC3-shKLF4 and LNCaP-shKLF4 cells was higher than that in PC3-con and LNCaP-con cells (P <0.01) N-cadherin, Zinc finger E-box-binding homeobox 1 (Zeb1), Snail1, Waveform in PC3-shKLF4 and LNCaPshKLF4 cells The expression levels of vimentin (Vim) and matrix metallopeptidase 1 (MMP1) mRNA were lower in PC3-con and LNCaP-con cells than those in control group (all P <0.05). The expression of E-cad in PC3-shKLF4 and LNCaP-shKLF4 cells was higher than that in PC3-con and LNCaP-con cells (all P <0.05), while N-cad in PC3-shKLF4 and LNCaP-shKLF4 cells , And the expression levels of Zeb1, Snail1, Vim and MMP1 were lower than that of PC3-con and LNCaP-con cells in control group (all P <0.05). The migration and invasion ability of PC3-shKLF4 and LNCaP-shKLF4 cells were lower than that of PC3-con and LNCaP-con cells respectively (P <0.01, P <0.05). Conclusion: Down-regulation of KLF4 expression in prostate cancer cells can promote the expression of epithelial-related genes and inhibit the expression of stromal-related genes, thereby inhibiting the migration and invasion of prostate cancer cells in vitro.