鸦胆子油乳对戈舍瑞林去势抵抗性前列腺癌PC3细胞株化疗敏感性的影响

Effect of Brucea javanica oil emulsion on chemotherapeutic sensitivity of goserelin castration resistant prostate cancer PC3 cell line

目的探讨质量分数10%鸦胆子油乳(Brucea javanica oil emulsion,BJOE)对戈舍瑞林去势抵抗性前列腺癌PC3细胞株化疗敏感性的影响及机制。方法对数生长期戈舍瑞林去势抵抗性前列腺癌PC3细胞株分为空白组、BJOE组、多西他赛组、多西他赛+BJOE组,细胞培养液中分别加入磷酸盐缓冲液、质量分数10%BJOE、0.6μg/L多西他赛、质量分数10%BJOE+0.6μg/L多西他赛进行干预。培养24、48、72h时,采用MTT法检测4组细胞增殖抑制率;培养48h时,采用流式细胞术检测4组细胞凋亡率,采用Hoechst 33258染色观察细胞形态学变化,采用实时荧光定量PCR法检测4组细胞B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2associated X protein,Bax)、caspase-3mRNA相对表达量,采用Western blot法检测4组细胞Bcl-2、Bax和caspase-3蛋白相对表达量。结果培养24、48、72h时,多西他赛+BJOE组、多西他赛组、BJOE组细胞增殖抑制率依次降低(P<0.05),3组细胞增殖抑制率均随时间延长而增高(P<0.05);培养48h时,空白组细胞核圆形或椭圆形,细胞质分布均匀;BJOE组、多西他赛组、多西他赛+BJOE组部分细胞呈不规则状态,染色质固缩、体积缩小,部分出现蓝色凋亡小体,其中多西他赛+BJOE组凋亡现象最明显;培养48h时,多西他赛+BJOE组、多西他赛组、BJOE组、空白组细胞凋亡率[(28.71±3.15)%、(18.85±2.03)%、(14.62±1.57)%、(9.10±1.02)%]依次降低(P<0.05),Bcl-2mRNA和蛋白相对表达量依次增高(P<0.05),Bax、caspase-3mRNA和蛋白相对表达量依次降低(P<0.05)。结论质量分数10%BJOE与多西他赛联用可抑制戈舍瑞林去势抵抗性前列腺癌细胞增殖,促进细胞凋亡,增强细胞对多西他赛化疗敏感性,可能与下调Bcl-2mRNA和蛋白表达,上调Bax、caspase-3mRNA和蛋白表达有关。

Objective To investigate the effect of 10%Brucea javanica oil emulsion(BJOE)on chemotherapy sensitivity of goserelin castration resistant prostate cancer PC3cell line.Methods The castration resistant prostate cancer PC3cell lines in logarithmic growth phase were divided into blank group,BJOE group,docetaxel group and docetaxel+BJOE group,added with phosphate buffer,10%BJOE,0.6μg/L docetaxel,and 10%BJOE+0.6μg/L docetaxel respectively to the cell culture medium for intervention.After culture for 24,48and 72h,MTT method was used to detect the inhibition rate of cell proliferation.After culture for 48h,the apoptosis rate was detected by flow cytometry.Hoechst 33258staining was used to observe the morphological changes.The expressions of B-cell lymphoma-2(Bcl-2),Bcl-2associated X protein(Bax)and caspase-3 mRNA were detected by real-time quantitative PCR.And the relative expressions of Bcl-2,Bax and caspase-3proteins were detected by Western blot.Results After culture for 24,48and 72h,the inhibition rate of cell proliferation decreased gradually in turn in docetaxel+BJOE group,docetaxel group and BJOE group(P<0.05),and increased with the time in three groups(P<0.05).After culture for 48h,the nucleus in blank group was round or oval and the cytoplasm was evenly distributed.In BJOE group,docetaxel group and docetaxel+BJOE group,some cells showed irregular,chromatin pyknosis and volume reduction,and some cells showed blue apoptotic bodies which was the most obvious in docetaxel+BJOE group.After culture for 48h,the apoptosis rate decreased gradually in turn in doxycycline+BJOE group,doxycycline group,BJOE group and blank group((28.71±3.15)%,(18.85±2.03)%,(14.62±1.57)%,(9.10±1.02)%)(P<0.05),the relative expressions of Bcl-2mRNA and protein increased gradually in turn(P<0.05),and the relative expressions of Bax and caspase-3mRNA and protein decreased gradually in turn(P<0.05).Conclusion The combination of 10%BJOE and docetaxel inhibits the proliferation of castration resistant prostate cancer cells,promotes the cell apoptosis and enhances its chemosensitivity to docetaxel,which might be correlated with the down-regulation of Bcl-2mRNA and protein,and up-regulation of Bax and caspase-3mRNA and protein.