目的:探讨健脾化瘀方对肝癌细胞Smad7蛋白及上皮间质转移标志蛋白E-cadherin/N-cadherin表达水平及侵袭转移能力的影响。方法:分别以不同浓度(20%,高浓度组;10%,低浓度组)健脾化瘀法中药含药血清及空白对照血清(空白对照组)培养肝癌细胞...目的:探讨健脾化瘀方对肝癌细胞Smad7蛋白及上皮间质转移标志蛋白E-cadherin/N-cadherin表达水平及侵袭转移能力的影响。方法:分别以不同浓度(20%,高浓度组;10%,低浓度组)健脾化瘀法中药含药血清及空白对照血清(空白对照组)培养肝癌细胞MHCC97-H,q PCR检测细胞Smad7、Snail、Zeb1、Zeb2 m RNA水平,Western blot检测Smad7、Snail、E-cadherin和N-cadherin蛋白表达水平。划痕法和Transwell小室检测肝癌细胞侵袭/迁移能力。结果:高浓度组Smad7 m RNA水平高于空白对照组(P<0.05),Snail、Zeb2 m RNA水平低于空白对照组(P<0.05)。与空白对照组比较,高浓度组48h的Snail、N-cadherin蛋白表达量明显降低,Smad7、E-cadherin蛋白表达量明显升高(P<0.05)。低浓度组48h的E-cadherin升高,N-cadherin降低(P<0.05)。划痕法结果显示,与空白对照组比较,高、低浓度组肝癌细胞的划痕愈合较慢(P<0.05,P<0.01)。Transwell显示,与空白对照组比较,高、低浓度组MHCC97-H细胞的迁移受抑制(P<0.05)。结论:健脾化瘀法中药能抑制肝癌细胞侵袭转移,可能与上调Smad7蛋白含量及抑制上皮间质转化有关。展开更多
OBJECTIVE:To examine the influence of SaponinⅠfrom Shuitianqi(Rhizoma Schizocapasae Plantagineae)(SSPHⅠ)on hepatocellular carcinoma(HCC)metastasis,and to elucidate the underlying mechanism.METHODS:The intrahepatic m...OBJECTIVE:To examine the influence of SaponinⅠfrom Shuitianqi(Rhizoma Schizocapasae Plantagineae)(SSPHⅠ)on hepatocellular carcinoma(HCC)metastasis,and to elucidate the underlying mechanism.METHODS:The intrahepatic metastasis Bagg's Albino/c(BALB/c)mouse model was established with human hepatocellular carcinomas(HepG2)cells,then treated with normal saline(once per day),cisplatin(2 mg/kg,once every 2 d),and SSPHⅠ(25,50,and 75 mg/kg,once per day).Then,we assessed alterations in the hepatic pathology and target protein expressions in the intrahepatic metastasis BALB/c mouse model using a series of molecular biology techniques.RESULTS:Based on our analysis,SSPHⅠsignificantly alleviated hepatocyte necrosis and tumor cells infiltration.Moreover,SSPHⅠsuppressed extracellular matrix(ECM)degradation and angiogenesis via a decrease in matrix etalloproteinase-2(MMP-2),MMP-9,CD31,CD34,and vascular endothelial growth factor(VEGF)levels.Furthermore,SSPHⅠrepressed invasion and metastasis by suppressing the transforming growth factor-β1(TGF-β1)/Smad7 axis and epithelial-mesenchymal transition(EMT),as evidenced by the scarce TGF-β1,Ncadherin,and Vimentin expressions,and elevated Smad7 and E-cadherin expressions.CONCLUSION:The SSPHⅠ-mediated negative regulation of the TGF-β1/Smad7 axis and EMT are critical for the inhibition of HCC invasion and metastasis.展开更多
文摘目的:探讨健脾化瘀方对肝癌细胞Smad7蛋白及上皮间质转移标志蛋白E-cadherin/N-cadherin表达水平及侵袭转移能力的影响。方法:分别以不同浓度(20%,高浓度组;10%,低浓度组)健脾化瘀法中药含药血清及空白对照血清(空白对照组)培养肝癌细胞MHCC97-H,q PCR检测细胞Smad7、Snail、Zeb1、Zeb2 m RNA水平,Western blot检测Smad7、Snail、E-cadherin和N-cadherin蛋白表达水平。划痕法和Transwell小室检测肝癌细胞侵袭/迁移能力。结果:高浓度组Smad7 m RNA水平高于空白对照组(P<0.05),Snail、Zeb2 m RNA水平低于空白对照组(P<0.05)。与空白对照组比较,高浓度组48h的Snail、N-cadherin蛋白表达量明显降低,Smad7、E-cadherin蛋白表达量明显升高(P<0.05)。低浓度组48h的E-cadherin升高,N-cadherin降低(P<0.05)。划痕法结果显示,与空白对照组比较,高、低浓度组肝癌细胞的划痕愈合较慢(P<0.05,P<0.01)。Transwell显示,与空白对照组比较,高、低浓度组MHCC97-H细胞的迁移受抑制(P<0.05)。结论:健脾化瘀法中药能抑制肝癌细胞侵袭转移,可能与上调Smad7蛋白含量及抑制上皮间质转化有关。
基金National Natural Science Foundation of China,a New Anti-cancer Plant drug,SaponinⅠfrom Shuitianqi(Rhizoma Schizocapasae Plantagineae),against Invasion and Metastasis of Non-small Cell Lung Cancer and Reversing Tyrosine Kinase Inhibitors Resistance basing on Human Growth Factor/c-Mesenchymal to Epithelial Transition Factor Pathway and its Molecular Mechanism of Regulating Epithelial-Mesenchymal Transition(No.8164062)the Natural Science Foundation of Guangxi Province,Study on the Antihepatic Fibrosis Mechanism of Saponins from Shuitianqi(Rhizoma Schizocapasae Plantagineae)based on Transforming Growth Factor-β/Smad Signaling Pathway(No.2019GXNSFAA245075)。
文摘OBJECTIVE:To examine the influence of SaponinⅠfrom Shuitianqi(Rhizoma Schizocapasae Plantagineae)(SSPHⅠ)on hepatocellular carcinoma(HCC)metastasis,and to elucidate the underlying mechanism.METHODS:The intrahepatic metastasis Bagg's Albino/c(BALB/c)mouse model was established with human hepatocellular carcinomas(HepG2)cells,then treated with normal saline(once per day),cisplatin(2 mg/kg,once every 2 d),and SSPHⅠ(25,50,and 75 mg/kg,once per day).Then,we assessed alterations in the hepatic pathology and target protein expressions in the intrahepatic metastasis BALB/c mouse model using a series of molecular biology techniques.RESULTS:Based on our analysis,SSPHⅠsignificantly alleviated hepatocyte necrosis and tumor cells infiltration.Moreover,SSPHⅠsuppressed extracellular matrix(ECM)degradation and angiogenesis via a decrease in matrix etalloproteinase-2(MMP-2),MMP-9,CD31,CD34,and vascular endothelial growth factor(VEGF)levels.Furthermore,SSPHⅠrepressed invasion and metastasis by suppressing the transforming growth factor-β1(TGF-β1)/Smad7 axis and epithelial-mesenchymal transition(EMT),as evidenced by the scarce TGF-β1,Ncadherin,and Vimentin expressions,and elevated Smad7 and E-cadherin expressions.CONCLUSION:The SSPHⅠ-mediated negative regulation of the TGF-β1/Smad7 axis and EMT are critical for the inhibition of HCC invasion and metastasis.
