基于AR通路及Wnt通路探讨固本清源方联合雄激素剥夺治疗去势抵抗性前列腺癌的机制

Mechanism of Guben Qingyuan Formula combined with androgen deprivation therapy inhibits castration-resistant prostate cancer by regulating the ARpathway and Wntpathway

目的:从前列腺癌AR通路及肿瘤干细胞Wnt通路探讨固本清源方联合雄激素剥夺(ADT)治疗控制去势抵抗性前列腺癌(CRPC)的作用机制。方法:建立CRPC荷瘤小鼠模型,采用RT-PCR、WesternBlot检测固本清源方联合ADT治疗肿瘤组织中AR、NKX3.1、PSA的表达;分选前列腺癌干细胞,观察固本清源方联合ADT治疗对接种前列腺癌干细胞荷瘤小鼠的抑瘤率,采用WesternBlot检测肿瘤组织中Wnt、β-catenin的表达。结果:中、高剂量固本清源方联合ADT治疗组AR、PSAmRNA及蛋白表达水平下调,而NKX3.1mRNA及蛋白表达水平上调,与ADT组治疗比较差异有统计学意义(P<0.05)。接种前列腺癌干细胞荷瘤小鼠较接种普通CRPC细胞荷瘤小鼠肿瘤显著增大(P<0.05)。低、中、高剂量固本清源方联合ADT治疗对前列腺癌干细胞荷瘤小鼠的抑瘤率分别为24.58%、36.67%、40.24%,与ADT组比较差异有统计学意义(P<0.05),进一步研究显示低、中、高剂量固本清源方联合ADT治疗能下调Wnt、β-catenin的蛋白表达水平,与ADT组比较差异有统计学意义(P<0.05)。结论:固本清源方联合ADT治疗CRPC作用机制与调控前列腺癌AR通路及肿瘤干细胞Wnt/β-catenin通路有关。

Objective:To explore the mechanism of Guben Qingyuan Formula combined with androgen deprivation therapy in the control of castration-resistant prostate cancer from the AR pathway of prostate cancer and the Wnt pathway of cancer stem cells.Methods:After establishing a castration-resistant prostate cancer tumor-bearing mouse model,RT-PCR and Western Blot were used to detect the expressions of AR,NKX3.1 and PSA in tumor tissues of Guben Qingyuan Formula combined with androgen deprivation therapy.Prostate cancer stem cells were sorted,and the tumor inhibition rate of Guben Qingyuan Formula combined with androgen deprivation therapy on prostate cancer stem cell-bearing mice was calculated.Then,the expressions of Wnt andβ-catenin in tumor tissues were detected by Western Blot.Results:The mRNA and protein expression levels of AR and PSA in the middle and high doses of Guben Qingyuan Formula combined with androgen deprivation therapy were significantly down-regulated,and NKX3.1 was significantly up-regulated,which was significantly different from androgen deprivation therapy(P<0.05).The tumors of tumor-bearing mice inoculated with prostate cancer stem cells were larger than those of castration-resistant prostate cancer-bearing mice,and there was a statistically significant difference in tumor mass between the two groups(P<0.05).The tumor inhibition rates of the low,medium and high-dose Guben Qingyuan Formula combined with androgen deprivation therapy groups were 24.58%,36.67%,and 40.24%,respectively,which were significantly different from those of the androgen deprivation therapy group alone(P<0.05).The study showed that low,medium and high-dose Guben Qingyuan Formula combined with androgen deprivation therapy group could down-regulate the protein expression levels of Wnt andβ-catenin,which were significantly different from those of the androgen deprivation therapy group alone(P<0.05).Conclusion:The mechanism of Guben Qingyuan Formula combined with androgen deprivation therapy in the control of castration-resistant prostate cancer is related to the regulation of the classical AR pathway and the Wnt/β-catenin pathway of cancerstem cells.