隐丹参酮对前列腺癌DU145细胞中异黏蛋白表达的影响

Crypotanshione reduces the expression of metadherin in DU145 prostate cancer cells

目的:探讨隐丹参酮对前列腺癌DU145细胞增殖及细胞凋亡的作用,并初步探讨隐丹参酮对DU145细胞中异黏蛋白(MTDH)表达及下游PI3K/AKT信号通路的影响。方法:四氮唑蓝(MTT)比色法检测不同浓度隐丹参酮分别作用DU145细胞24、48、72 h后对细胞的生长抑制作用;原位末端转移酶标记(TUNEL)法检测细胞凋亡情况;Western印迹检测不同浓度隐丹参酮及作用不同时间对DU145细胞中MTDH蛋白的表达影响;RT-PCR技术检测隐丹参酮分别作用DU145细胞12、24、48 h后细胞中MTDH mRNA的表达情况;Western印迹检测隐丹参酮作用DU145细胞48 h后细胞中MTDH、AKT、p-AKT、Bcl-2蛋白的表达情况。结果:隐丹参酮能够明显抑制DU145细胞增殖,且抑制效应呈剂量和时间依赖性(P<0.05);以10μmol/L隐丹参酮作用DU145细胞24、48、72 h后细胞凋亡率分别为(29.42±4.51)%、(55.07±5.67)%和(70.84±4.66)%,明显高于对照组(3.1±2.48)%(P<0.05)。Western印迹和RT-PCR结果显示,隐丹参酮可在转录和翻译水平下调MTDH的表达(P<0.05),抑制AKT信号通路和抗凋亡蛋白Bcl-2的表达(P<0.05)。结论:隐丹参酮可抑制前列腺癌DU145细胞的增殖,促进细胞凋亡,其机制可能是通过下调MTDH表达,抑制其下游PI3K/AKT信号通路。

Objective ： To investigate the effects of crypotanshinone （CPT） on the proliferation and apoptosis of DU145 prostate cancer cells as well as on the metadherin expression and the downstream PI3K/AKT signaling pathway in the DU145 cells. Methods ： We treated DU145 prostate cancer cells with different concentrations of CPT for 24, 48, and 72 hours followed by evaluation of the proliferation and apoptosis of the cells by MTT assay and TUNEL, respectively. We determined the expressions of metadherin protein and mRNA in the DU145 cells by Western blot and RT-PCR respectively at different time points after CPT treatment. We also detected the expressions of the proteins metadherin, AKT, p-AKT, and Bcl-2 in the CPT-treated DU145 cells at 48 hours. Results ： Clef significantly inhibited the proliferation of the DU145 cells in a dose- and time-dependent manner （ P 〈 0.05 ）. After treatment with 10 μmol/L CPT for 24, 48, and 72 hours, the apoptosis rates of the DU145 cells were （29.42 ±4.51）, （55.07±5.67） and （70.84 ± 4.66） %, respectively, significantly higher than （ 3. 1 ± 2.48 ） % in the control group （ P 〈 0.05 ）. The expression of metadherin was remarkably downregulated at the transcription and translation levels （ P 〈 0.05 ） and the expressions of the AKT signaling pathway and the Bcl-2 protein were markedly inhibited in the DU145 cells after treated with 10μmol/L CPT for 48 hours （P 〈 0.05 ）. Conclusion ： CPT can inhibit the proliferation and induce the apoptosis of DU145 prostate cancer cells, which may be associated with its suppression of the downstream PI3K/AKT signaling pathway by reducing the expression of metadherin in the DU145 cells.