从肿瘤干细胞Wnt通路探讨固本清源方药联合雄激素剥夺治疗控制去势抵抗性前列腺癌

Study on Guben Qingyuan Prescription(固本清源方)Combined with Androgen Deprivation Therapy to Control Castration-Resistant Prostate Cancer Based on Wnt Pathway of Cancer Stem Cells

目的从肿瘤干细胞Wnt通路探讨固本清源方药联合雄激素剥夺治疗控制去势抵抗性前列腺癌的作用机制。方法采用流式细胞术从前列腺癌PC-3细胞中分选出CD44+CD133+PC-3细胞并验证其肿瘤干细胞特性。将36只雄性裸鼠随机分为A~F组,每组6只,除A组接种前列腺癌PC-3细胞外,其他各组接种前列腺癌干细胞;除A、B组均给予氯化钠溶液外,其余组均给予醋酸戈舍瑞林缓释植入剂0.36 mg·kg^(-1),腹腔注射1次/25 d;比卡鲁胺片5 mg·kg^(-1),灌胃1次/d;其中D-F组在上述基础上进一步分别给予2.5 g·kg^(-1)、25 g·kg^(-1)、50 g·kg^(-1)固本清源方药,灌胃2次/d;观察荷瘤小鼠肿瘤的生长情况,连续给药32 d后剥离瘤组织,计算肿瘤抑制率;采用Western blot法分别检测肿瘤组织中Wnt、β-catenin的蛋白表达。结果从PC-3细胞中分选出的CD44+CD133+PC-3细胞亚群具有前列腺癌干细胞特性;低、中、高剂量固本清源方联合雄激素剥夺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%,与单纯雄激素剥夺治疗组比较差异具有统计学意义(P<0.05),进一步研究显示低、中、高剂量固本清源方联合雄激素剥夺治疗能下调Wnt、β-catenin的蛋白表达水平,与雄激素剥夺治疗组比较差异有统计学意义(P<0.05)。结论固本清源方联合雄激素剥夺治疗能够抑制前列腺癌干细胞荷瘤小鼠肿瘤生长,其作用机制与调控前列腺癌干细胞Wnt/β-catenin信号通路有关。

Objective To investigate the efficacy and mechanism of the Guben Qingyuan Prescription(固本清源方)combined with androgen deprivation therapy to control castration-resistant prostate cancer through the Wnt pathway of tumor stem cells.Methods CD44+CD133+PC-3 cells were sorted out from PC-3 cells by flow cytometry and their tumor stem cell properties were verified.Thirty-six male nude mice were randomly divided into 6 groups(groups A-F).All groups were inoculated with prostate cancer stem cells except group A which was inoculated with prostate cancer PC-3 cells.All groups were given goserelin(0.36 mg·kg^(-1))by intraperitoneal injection every 25 days and bicalutamide(5 mg·kg^(-1))by gavage once a day except group A and B which were both given the equal amounts of saline.Group D-F were further given Guben Qingyuan Prescription(2.5 g·kg^(-1),25 g·kg^(-1),50 g·kg^(-1))by gavage twice a day,respectively.The growth of tumors in the tumor-bearing mice was observed,and the tumor tissues were peeled off after 32 days of continuous administration to calculate the tumor inhibition rate.The protein expressions of Wnt andβ-catenin in tumor tissues were detected separately by Western blot.Results A subpopulation of CD44+CD133+PC-3 cells isolated from PC-3 cells had prostate cancer stem cell characteristics.The tumor suppression rates of low,medium and high doses of Guben Qingyuan Prescription combined with androgen deprivation treatment on prostate cancer stem cell-bearing mice were 24.58%,36.67%and 40.24%,respectively,which were statistically different from the androgen deprivation treatment group(P<0.05).Further study showed that low,medium and high doses of Guben Qingyuan Prescription combined with androgen deprivation treatment could down-regulate the protein expression levels of Wnt andβ-catenin,which were statistically different from the androgen deprivation treatment group(P<0.05).Conclusion The combination of androgen deprivation therapy with Guben Qingyuan Prescription may control the tumor growth in prostate cancer stem cell-bearing mice by regulating the Wn/β-catenin signaling pathway.