益肾通癃颗粒调控Wnt/β-catenin信号通路对人前列腺癌PC3细胞荷瘤裸鼠的干预作用

Interventional effects of Yishen Tonglong Granules on regulating the Wnt/β-catenin signaling pathway in nude mice bearing human prostate cancer cell PC3 tumor

目的观察益肾通癃颗粒对人前列腺癌PC3细胞荷瘤裸鼠Wnt/β-catenin信号通路的干预作用。方法利用人前列腺癌骨转移细胞PC3建立前列腺癌荷瘤裸鼠模型,将其随机分为模型对照组、阳性药物组、益肾通癃颗粒低剂量组、益肾通癃颗粒中剂量组、益肾通癃颗粒高剂量组和联合用药组,给予相应的药物干预,连续4周。给药结束后处死动物获取皮下移植瘤瘤体组织样本,分别进行HE染色观察病理变化,免疫组化检测细胞增殖情况,TUNEL检测细胞凋亡情况,Western blot及免疫组织化学法检测Wnt信号通路相关基因蛋白的表达水平。结果益肾通癃颗粒可以有效改善荷瘤裸鼠皮下移植瘤瘤体组织的病理改变,可以显著抑制肿瘤细胞的增殖,促进细胞凋亡的发生,药效呈浓度依赖性。益肾通癃颗粒还可以下调Wnt/β-catenin信号通路相关基因Wnt1、Wnt3a、β-catenin、APC蛋白的表达(P<0.01),上调Wnt/β-catenin信号通路相关基因GSK-3β蛋白的表达(P<0.01),降低p-GSK-3β蛋白的活性(P<0.01),药效与剂量呈正相关性,与Wnt信号通路阻断剂ICG联合时效果最佳(P<0.01)。结论益肾通癃颗粒能够抑制前列腺癌细胞的增殖,促进其凋亡,其干预作用可能与其阻断Wnt/β-catenin信号通路激活密切相关。

Objective To observe the intervention effects of Yishen Tonglong Granules(YSTLG)on the Wnt/β-catenin signaling pathway in nude mice bearing human prostate cancer cell PC3 tumor.Methods The models of nude mice bearing prostate cancer tumor were prepared using human prostate cancer bone metastasis cell PC3 and were randomized into model control,positive drug,and low-,medium-,and high-dose YSTLG,and combination drug groups.Corresponding drug interventions were given for four consecutive weeks.After administration,the animals were sacrificed to obtain subcutaneous transplanted tumor tissue samples.HE staining was performed to observe pathological changes,immunohistochemistry was used to check cell proliferation,TUNEL was used to check cell apoptosis,and Western blot and immunohistochemistry were used to check the expression levels of Wnt signaling pathway-related gene proteins.Results YSTLG can effectively relieve the pathological changes of subcutaneous transplanted tumor tissue in nude mice bearing tumor,significantly inhibit the proliferation of tumor cells,and promote cell apoptosis with concentration-dependent effects.YSTLG can also downregulate the APC protein expressions of Wnt/β-catenin signaling pathway related genes including Wnt1,Wnt3a,andβ-catenin(P<0.01),upregulate the GSK-3βprotein expression of Wnt/β-catenin signaling pathway related gene(P<0.01),and reduce the activity of p-GSK-3βprotein(P<0.01).The correlation between drug efficacy and dosage was positive,and the best effects are achieved when YSTLG was combined with the Wnt signaling pathway blocker ICG(P<0.01).Conclusion YSTLG could inhibit the proliferation of prostate cancer cells,promote their apoptosis,which are likely to be achieved by blocking Wnt/β-catenin signaling pathway.