氯喹抑制高糖环境下前列腺癌PC3细胞增殖机制研究

Inhibition of chloroquine on the proliferation of prostate cancer cell line PC3 in high glucose condition

目的:研究氯喹是否能通过自噬和活性氧(ROS)通路抑制高糖诱导的前列腺癌PC3细胞的增殖。方法:PC3细胞培养后分为4组(正常糖组、高糖组、正常糖+氯喹组、高糖+氯喹组)予相应干预。运用CCK-8实验检测细胞的增殖能力;Western blot检测自噬相关蛋白LC3的表达;DCFH-DA活性氧荧光探针检测细胞中ROS水平;JC-1法检测细胞线粒体膜电位的去极化。结果:CCK-8实验结果表明氯喹能够显著抑制高糖环境下PC3细胞的增殖。Western blot实验结果显示氯喹会使PC3细胞LC3-Ⅱ/Ⅰ比值升高,说明细胞自噬显著被抑制。DCFH-DA探针检测结果表明,氯喹能够显著上调高糖环境下PC3细胞中ROS水平。JC-1检测结果表明,氯喹能够诱导高糖环境下PC3细胞中线粒体膜电位去极化。结论:氯喹通过自噬和ROS通路抑制高糖环境下前列腺癌细胞的增殖。

Objective:To investigate whether chloroquine can inhibit the proliferation of prostate cancer cell line PC3 in high glucose condition via autophagy and reactive oxygen species(ROS)pathways.Methods:PC3 cells were divided into four groups:normal glucose group,high glucose group,normal glucose+chloroquine group and high glucose+chloroquine group.CCK-8 assay was applied to assess the effect of proliferation.The protein expression of LC3 related to autophagy was detected with Western blot.ROS levels were detected using DCFH-DA staining in PC3 cells.JC-1 staining was performed to investigate the depolarization of mitochondrial membrane in PC3 cells.Results:CCK-8 assay showed that the proliferation of PC3 cells were significantly inhibited by chloroquine in high glucose condition.Western blot assay indicated that the LC3-Ⅱ/Ⅰexpression of PC3 cells were elevated by chloroquine in high glucose condition,which illustrated that chloroquine inhibited the autophagy of PC3 cells.DCFH-DA staining assay showed that the ROS levels in PC3 cells were up-regulated by chloroquine in high glucose condition.Furthermore,results of JC-1 assay demonstrated that chloroquine induced the depolarization of mitochondrial membrane of PC3 cells in high glucose condition.Conclusion:Chloroquine can inhibit the proliferation of prostate cancer cell line PC3 in high glucose condition via autophagy and ROS pathways.