DLC-1通过Rho通路抑制耐恩杂鲁胺前列腺癌细胞的增殖与侵袭能力

Deleted in liver cancer-1 inhibits the proliferation and invasion of enzalutamide-resistant prostate cancer cells via the Rho pathway

摘要

目的 探索肝癌缺失因子1（deleted in liver cancer 1，DLC-1）对耐恩杂鲁胺前列腺癌（prostate cancer，PCa）细胞增殖，侵袭和凋亡的影响以及作用机制。 方法 体外以不同浓度的恩杂鲁胺药物培养人前列腺癌lncap及C4-2细胞株6个月以上，构建恩杂鲁胺耐药细胞系lncap-R，C4-2-R。CCK-8法检测其IC₅₀，验证恩杂鲁胺耐药株是否构建成功。利用慢病毒转染lncap-R，C4-2-R，以敲低和过表达DLC-1，采用克隆实验、Transwell实验和流式细胞术检测DLC-1对耐恩杂鲁胺的前列腺癌细胞的增殖、侵袭和凋亡的影响。 结果 与对照组相比，过表达组DLC-1显著抑制了2个耐药株的增殖和侵袭进程。机制上，WB实验结果表明DLC-1表达与2个耐药株中Rho蛋白的表达呈负相关。Rho抑制剂rhosin逆转了因敲低DLC-1后lncap-R细胞凋亡减少的情况。 结论 DLC-1可以下调耐药株中的Rho蛋白的表达，并抑制耐药株的增殖和侵袭。

Abstract

Objective To investigate the effect of deleted in liver cancer-1（DLC-1） on the proliferation，invasion，and apoptosis of enzalutamide-resistant prostate cancer cells and its mechanism of action. Methods Hμmol/Lan prostate cancer lncap and C4-2 cell lines were cultured in vitro with different concentrations of enzalutamide for more than 6 months，and enzalutamide-resistant cell lines were constructed as lncap-R and C4-2-R. IC₅₀ was measured by CCK-8 assay to verify the successful construction of enzalutamide-resistant cell lines. lncap-R and C4-2-R were transfected by lentivirus to achieve the knockdown and overexpression of DLC-1，and colony formation assay，Transwell assay，and flow cytometry were used to observe the effect of DLC-1 on the proliferation，invasion，and apoptosis of enzalutamide-resistant prostate cancer cells. Results Compared with the control group，the overexpression group showed that DLC-1 significantly inhibited the proliferation and invasion of the two enzalutamide-resistant cell lines. As for mechanism of action，Western blot showed that the expression of DLC-1 was negatively correlated with the expression of Rho protein in both enzalutamide-resistant cell lines. The Rho inhibitor rhosin reversed the reduction in the apoptosis of lncap-R cells due to knockdown of DLC-1. Conclusion DLC-1 can downregulate the expression of Rho protein in drug-resistant cell lines and inhibit their proliferation and invasion.

关键词

肝癌缺失因子1 / 前列腺癌 / 恩杂鲁胺 / Rho蛋白 / 增殖 / 侵袭

Key words

deleted in liver cancer-1 / prostate cancer / enzalutamide / Rho protein / proliferation / invasion

基金资助

国家自然科学基金资助项目(81802543)