隐丹参酮调节TGF-β1/Smad3介导的上皮间质转化对抑制前列腺癌小鼠肿瘤生长的影响

Cryptotanshinone Regulates TGF-β1/Smad3 Mediated Epithelial-mesenchymal Transformation and Inhibits Tumor Growth in Prostate Cancer Mice

目的 探讨隐丹参酮调节转化生长因子β1(TGF-β1)/Smad3信号通路介导的上皮间质转化(EMT)对前列腺癌小鼠肿瘤生长的影响。方法 50只小鼠右侧腋皮下注射PC-3细胞悬液建立前列腺癌移植瘤小鼠模型,将造模成功的44只小鼠随机分为模型组(10只)、隐丹参酮低剂量组(12只)、隐丹参酮高剂量组(12只)和多西紫杉醇组(10只),另取10只作为正常组。隐丹参酮低和高剂量组小鼠分别腹腔注射隐丹参酮20和40 mg/kg,1次/2 d,多西紫杉醇组小鼠腹腔注射多西紫杉醇0.25 mg/mL,1次/7 d,连续给药42 d。测定肿瘤体积和抑瘤率,ELISA检测血清中白介素(IL)-6、肿瘤坏死因子α(TNF-α)和前列腺特异性抗原(PSA)水平以及超氧化物歧化酶(SOD)活性和丙二醛(MDA),qRT-PCR法检测上皮钙粘蛋白(E-cad)、神经钙粘蛋白(N-cad)和波形蛋白(Vimentin) mRNA水平,蛋白印迹法检测E-cad、N-cad、Vimentin、TGF-β1和p-Smad3蛋白相对表达量。结果 与模型组比较,隐丹参酮低、高剂量组和多西紫杉醇组肿瘤体积减小,IL-6、TNF-α和PSA、MDA水平降低,SOD活性以及E-cad mRNA水平和蛋白相对表达量升高,N-cad和Vimentin mRNA水平和蛋白相对表达量降低,TGF-β1和p-Smad3蛋白相对表达量降低,其中各指标变化以多西紫杉醇组最为显著,其次是隐丹参酮高剂量组和隐丹参酮低剂量组(P<0.05)。结论 隐丹参酮可能通过抑制TGF-β1/Smad3信号通路逆转EMT过程,从而抑制前列腺癌小鼠肿瘤生长,降低炎症反应。

Objective To investigate the effect of cryptotanshinone on tumor growth in mice with prostate cancer by regulating transforming growth factorβ1(TGF-β1)/Smad3 signaling pathway mediated by epithelial-mesenchymal transformation(EMT).Methods 50 mice on the right side of axillary hypodermic PC-3 cell suspension prostate cancer transplanted tumor mice model is set up,building a successful 44 mice were randomly divided into model group(10),the cryptotanshinone low-dose group(12),cryptotanshinone high dose group(12)and docetaxel group(10),another 10 mice as normal group.Mice in the low-dose and high-dose cryptotanshinone groups were intraperitoneally injected with cryptotanshinone 20 and 40 mg/kg,once for 2 d,and mice in the docetaxel group were intraperitoneally injected with docetaxel 0.25 mg/mL,once for 7 d,for consecutive 42 d.Tumor volume and tumor inhibition rate were determined.Serum levels of interleukin(IL-6),tumor necrosis factorα(TNF-α)and prostate specific antigen(PSA)were detected by ELISA.Histopathological morphology of tumor tissue was observed by HE staining.MRNA levels of epithelial cadherin(E-cad),neurocadherin(N-cad)and Vimentin were detected by qRT-PCR,and relative protein expression levels of E-cad,N-cad,Vimentin,TGF-β1 and P-Smad3 were detected by western blot.Results Compared with model group,tumor volume decreased,MDA,IL-6,TNF-αand PSA levels decreased,activity of SOD and E-cad mRNA levels and protein relative expression increased,N-cad and Vimentin mRNA levels and protein relative expression decreased in low and high dose cryptotanshinone groups and docetaxel groups.The relative expression levels of TGF-β1 and p-smad3 protein were decreased,and the changes of each index were most significant in docetaxel group,followed by cryptotanshinone high-dose group and cryptotanshinone low-dose group(P<0.05).Conclusion Cryptotanshinone may reverse the EMT process by inhibiting TGF-β1/Smad3 signaling pathway,thereby inhibiting tumor growth and reducing inflammatory response in prostate cancer mice.