基于ROS/PI3K/Akt/mTOR信号通路探讨玫瑰花甲醇提取物对前列腺癌PC-3细胞凋亡的影响

目的：探讨玫瑰花甲醇提取物（RE）对前列腺癌PC-3细胞增殖和凋亡的影响及潜在作用机制。方法：采用CCK8法检测不同浓度RE对PC-3细胞增殖的影响；采用Hoechst染色法和Annexin V/PI双染法检测细胞凋亡；DCFH-DA检测细胞ROS水平；Western blot法检测Bax、Bcl-2、cleaved caspase-3、Cyt-C、PI3K、p-PI3K、Akt、p-Akt、mTOR、p-mTOR的蛋白表达。结果：RE呈浓度依赖性抑制前列腺癌PC-3细胞增殖，24 h和48 h后的IC₅₀值分别为31.30 mg·mL^-1和16.76 mg·mL^-1；RE能显著诱导PC-3细胞凋亡（P<0.05，P<0.01），且呈现浓度依赖性；RE可显著提高PC-3细胞ROS水平（P<0.05，P<0.01），且具有浓度依赖性；RE能显著上调PC-3细胞Bax、cleaved caspase-3、Cyt-C的蛋白表达及Bax/Bcl-2的比值（P<0.05，P<0.01），显著下调Bcl-2、p-PI3K、p-Akt、p-mTOR的蛋白表达（P<0.05，P<0.01），以上均呈现浓度依赖性。结论：本研究表明RE在体外有明显抑制前列腺癌细胞PC-3的增殖作用，并可能通过调控ROS/PI3K/Akt/mTOR信号通路诱导其凋亡，研究结果可为玫瑰花及其组方临床治疗前列腺癌提供实验依据。

Effects of apoptosis induced by methanolic extract of Rosa rugosa Thunb. in prostate cancer PC-3 cells based upon ROS/PI3K/Akt/mTOR signaling pathway

OBJECTIVE To explore the effects of methanolic extract of Rosa rugosa Thunb. (RE) on the proliferation and apoptosis of prostate cancer PC-3 cell and elucidate its potential mechanism.METHODS The effects of different concentrations of RE on cellular proliferation were detected by CCK8 assay. Apoptosis was detected by Hoechst staining and annexin V/PI double staining. The cellular level of reactive oxygen species (ROS) was measured by DCFH-DA. And Bax, Bcl-2, cleaved caspase-3, Cyt-C, PI3K, p-PI3K, Akt, p-Akt, mTOR and p-mTOR were detected by Western Blot.RESULTS RE suppressed cellular proliferation in a concentration-dependent manner and IC₅₀ values were 31.30 and 16.76 mg·mL^-1 at 24 h and 48 h. It could significantly induce cellular apoptosis in a concentration-dependent manner (P<0.05, P<0.01), elevate the cellular level of ROS in a concentration-dependent manner (P<0.05, P<0.01), up-regulate the protein expressions of Bax, cleaved caspase-3, Cyt-C, enhance the ratio of Bax/Bcl-2 (P<0.05, P<0.01) and down-regulate the protein expressions of Bcl-2, p-PI3K, p-Akt and p-mTOR in a concentration-dependent manner (P<0.05, P<0.01).CONCLUSION RE can significantly suppress the in vitro proliferation of prostate cancer PC-3 cell and induce apoptosis through regulating ROS/PI3K/Akt/mTOR signaling pathway. The results of the study may provide experimental rationales for clinical treatment of prostate cancer with Rosa rugosa Thunb. and its formula.