健脾化瘀方治疗原发性肝癌的实验研究
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摘要
研究目的:本研究旨在探讨:①健脾化瘀方体外对人肝癌细胞株SMMC-7721增殖的抑制;②健脾化瘀方体外对人肝癌细胞株SMMC-7721凋亡的影响;③健脾化瘀方体外对人肝癌细胞株SMMC-7721 ras/MAPK信号通路相关基因表达的影响。
研究方法:①采用MTT法,观察不同浓度的健脾化瘀方对人肝癌细胞株SMMC-7721增殖的影响及与药物作用时间的关系;②利用流式细胞仪,AnnexinV/PI双染色法,研究不同浓度健脾化瘀方对人肝癌细胞株SMMC-7721早期凋亡的诱导作用;③RT-PCR法研究健脾化瘀方对SMMC-7721 ras/MAPK信号传导通路相关基因的影响。
研究结果:①健脾化瘀方对人肝癌细胞株SMMC-7721增殖的抑制有一定的浓度依赖性:当药物浓度为0.625mg/ml时,无明显的抑制作用;当药物浓度在5mg/ml,对肝癌细胞株的增殖有明显的抑制作用,随着时间的增加抑制作用逐渐增强。②健脾化瘀方对人肝癌细胞SMMC-7721作用24H,有诱导人肝癌细胞株SMMC-7721凋亡的作用,并有一定的浓度依赖性。③健脾化瘀方对人肝癌细胞株SMMC-7721作用48H后,ras/MAPK信号通路上ERK、sos1、c-fos基因的表达较对照组低;MEK、Raf、ras、Grb2基因的表达与对照组比较,差异无统计学意义。
研究结论:①健脾化瘀方对人肝癌细胞株SMMC-7721的增殖具有抑制作用;②健脾化瘀方有诱导人肝癌细胞株SMMC-7721凋亡的作用;③健脾化瘀方能降低人肝癌细胞株SMMC-7721的ras/MAPK信号传导通路中ERK、sos1、c-fos基因的表达。
Objective:This study was designed to explore:①The inhibiting effect of Jianpi Huayu Fang in vitro on the proliferation of human hepatoma cell line SMMC-7721;②The effect of Jianpi Huayu Fang in vitro on the apoptosis of human hepatoma cell line SMMC-7721;③The effect of Jianpi Huayu Fang in vitro on human hepatoma cell line SMMC-7721 ras/MAPK signaling pathway-related gene expression.
Method:①MTT method was used to observe the effects of different concentrations of Jianpi Huayu Fang on human hepatoma cell line SMMC-7721 proliferation and the relationship between drug effect and time;②Flow cytometry,AnnexinV/PI double staining method was used to observe the early apoptosis induced by different concentrations of Jianpi Huayu Fang in human hepatoma cell line SMMC-7721;③RT-PCR method was used to observe the influence of Jianpi Huayu Fang on SMMC-7721 ras/MAPK signal transduction pathway related genes.
Results:①Jianpi Huayu Fang inhibited the proliferation of human hepatoma cell line SMMC-7721 with a certain degree of concentration-dependence:no significant inhibition was observed when the drug concentration was 0.625mg/ml;it significantly inhibited the proliferation of human hepatoma cell line when the drug concentration increased to 5mg/ml and the inhibition gradually increased with time.②Jianpi Huayu Fang applied on human hepatocellular carcinoma cell SMMC-7721 for 24H,induced the apoptosis of human hepatoma cell line SMMC-7721 with a certain degree of concentration-dependence.③Ras/MAPK signal transduction pathway ERK,SOS1,c-fos gene expression was decreased compared with control group after Jianpi Huayu Fang was applied on human hepatoma cell line SMMC-7721 for 48H; MEK,Raf,Ras,Grb2 gene expression showed no statistically significant difference compared with the control group.
Conclusions:①Jianpi Huayu Fang inhibited the proliferation of human hepatoma cell line SMMC-7721;②Jianpi Huayu Fang induced the apoptosis of human hepatoma cell line SMMC-7721;③Jianpi Huayu Fang affected the ras/MAPK signal transduction pathway ERK, sos1,c-fos gene expression of human hepatoma cell line SMMC-7721.
研究方法:①采用MTT法,观察不同浓度的健脾化瘀方对人肝癌细胞株SMMC-7721增殖的影响及与药物作用时间的关系;②利用流式细胞仪,AnnexinV/PI双染色法,研究不同浓度健脾化瘀方对人肝癌细胞株SMMC-7721早期凋亡的诱导作用;③RT-PCR法研究健脾化瘀方对SMMC-7721 ras/MAPK信号传导通路相关基因的影响。
研究结果:①健脾化瘀方对人肝癌细胞株SMMC-7721增殖的抑制有一定的浓度依赖性:当药物浓度为0.625mg/ml时,无明显的抑制作用;当药物浓度在5mg/ml,对肝癌细胞株的增殖有明显的抑制作用,随着时间的增加抑制作用逐渐增强。②健脾化瘀方对人肝癌细胞SMMC-7721作用24H,有诱导人肝癌细胞株SMMC-7721凋亡的作用,并有一定的浓度依赖性。③健脾化瘀方对人肝癌细胞株SMMC-7721作用48H后,ras/MAPK信号通路上ERK、sos1、c-fos基因的表达较对照组低;MEK、Raf、ras、Grb2基因的表达与对照组比较,差异无统计学意义。
研究结论:①健脾化瘀方对人肝癌细胞株SMMC-7721的增殖具有抑制作用;②健脾化瘀方有诱导人肝癌细胞株SMMC-7721凋亡的作用;③健脾化瘀方能降低人肝癌细胞株SMMC-7721的ras/MAPK信号传导通路中ERK、sos1、c-fos基因的表达。
Objective:This study was designed to explore:①The inhibiting effect of Jianpi Huayu Fang in vitro on the proliferation of human hepatoma cell line SMMC-7721;②The effect of Jianpi Huayu Fang in vitro on the apoptosis of human hepatoma cell line SMMC-7721;③The effect of Jianpi Huayu Fang in vitro on human hepatoma cell line SMMC-7721 ras/MAPK signaling pathway-related gene expression.
Method:①MTT method was used to observe the effects of different concentrations of Jianpi Huayu Fang on human hepatoma cell line SMMC-7721 proliferation and the relationship between drug effect and time;②Flow cytometry,AnnexinV/PI double staining method was used to observe the early apoptosis induced by different concentrations of Jianpi Huayu Fang in human hepatoma cell line SMMC-7721;③RT-PCR method was used to observe the influence of Jianpi Huayu Fang on SMMC-7721 ras/MAPK signal transduction pathway related genes.
Results:①Jianpi Huayu Fang inhibited the proliferation of human hepatoma cell line SMMC-7721 with a certain degree of concentration-dependence:no significant inhibition was observed when the drug concentration was 0.625mg/ml;it significantly inhibited the proliferation of human hepatoma cell line when the drug concentration increased to 5mg/ml and the inhibition gradually increased with time.②Jianpi Huayu Fang applied on human hepatocellular carcinoma cell SMMC-7721 for 24H,induced the apoptosis of human hepatoma cell line SMMC-7721 with a certain degree of concentration-dependence.③Ras/MAPK signal transduction pathway ERK,SOS1,c-fos gene expression was decreased compared with control group after Jianpi Huayu Fang was applied on human hepatoma cell line SMMC-7721 for 48H; MEK,Raf,Ras,Grb2 gene expression showed no statistically significant difference compared with the control group.
Conclusions:①Jianpi Huayu Fang inhibited the proliferation of human hepatoma cell line SMMC-7721;②Jianpi Huayu Fang induced the apoptosis of human hepatoma cell line SMMC-7721;③Jianpi Huayu Fang affected the ras/MAPK signal transduction pathway ERK, sos1,c-fos gene expression of human hepatoma cell line SMMC-7721.
引文
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