Objective: It is discussed whether saikosaponin A induces apoptosis of human hepatoma Huh7 cells is related to the change of autophagy level.Methods: The effects of different concentrations of SSA on proliferation and...Objective: It is discussed whether saikosaponin A induces apoptosis of human hepatoma Huh7 cells is related to the change of autophagy level.Methods: The effects of different concentrations of SSA on proliferation and apoptosis of Huh7 cells were detected by MTT and flow cytometry, and then constructed recombinant plasmid pEGFP-N1-LC3B and transfected into Huh7 cells. After intervened by SSA culture medium, the autophagy level was observed under confocal microscope. The expression of apoptosis proteins Bax, Bcl-2, PCNA and autophagy-related proteins LC3B, Beclin1, and Apg12-Apg5 were detected by Western Blot. Results: SSA can significantly inhibit the proliferation of Huh7 cells, promote apoptosis, increase the number of autophagy bodies in the cytoplasm, up-regulate the expression of Bax, LC3B-II, Beclin1, Apg12-Apg5 and down-regulate the expression of Bcl-2, PCNA. Conclusion:SSA induced apoptosis of Huh7 cells in vitro and upregulated the autophagy level.展开更多
目的检测抑制素在人肝癌细胞SMMC-7721及Huh-7中的基因表达,扩增抑制素全长编码序列。方法分别提取肝癌细胞SMMC-7721及Huh-7细胞总RNA,逆转录合成cDNA。使用Real time PCR,以GAPDH为内参检测抑制素的mRNA表达情况。采用RT-PCR方法扩增...目的检测抑制素在人肝癌细胞SMMC-7721及Huh-7中的基因表达,扩增抑制素全长编码序列。方法分别提取肝癌细胞SMMC-7721及Huh-7细胞总RNA,逆转录合成cDNA。使用Real time PCR,以GAPDH为内参检测抑制素的mRNA表达情况。采用RT-PCR方法扩增抑制素全长编码序列。结果 Real time PCR结果显示,SMMC-7721和Huh-7肝癌细胞株中均有抑制素的表达,且SMMC-7721细胞中抑制素表达量是Huh-7细胞中的6.652倍。RT-PCR方法扩增得到约835 bp的抑制素全长编码序列,条带特异,无非特异性扩增。结论人肝癌细胞株SMMC-7721中抑制素的表达量较高。展开更多
基金This study was supported by National Natural Science Foundation(81273745).
文摘Objective: It is discussed whether saikosaponin A induces apoptosis of human hepatoma Huh7 cells is related to the change of autophagy level.Methods: The effects of different concentrations of SSA on proliferation and apoptosis of Huh7 cells were detected by MTT and flow cytometry, and then constructed recombinant plasmid pEGFP-N1-LC3B and transfected into Huh7 cells. After intervened by SSA culture medium, the autophagy level was observed under confocal microscope. The expression of apoptosis proteins Bax, Bcl-2, PCNA and autophagy-related proteins LC3B, Beclin1, and Apg12-Apg5 were detected by Western Blot. Results: SSA can significantly inhibit the proliferation of Huh7 cells, promote apoptosis, increase the number of autophagy bodies in the cytoplasm, up-regulate the expression of Bax, LC3B-II, Beclin1, Apg12-Apg5 and down-regulate the expression of Bcl-2, PCNA. Conclusion:SSA induced apoptosis of Huh7 cells in vitro and upregulated the autophagy level.
文摘目的检测抑制素在人肝癌细胞SMMC-7721及Huh-7中的基因表达,扩增抑制素全长编码序列。方法分别提取肝癌细胞SMMC-7721及Huh-7细胞总RNA,逆转录合成cDNA。使用Real time PCR,以GAPDH为内参检测抑制素的mRNA表达情况。采用RT-PCR方法扩增抑制素全长编码序列。结果 Real time PCR结果显示,SMMC-7721和Huh-7肝癌细胞株中均有抑制素的表达,且SMMC-7721细胞中抑制素表达量是Huh-7细胞中的6.652倍。RT-PCR方法扩增得到约835 bp的抑制素全长编码序列,条带特异,无非特异性扩增。结论人肝癌细胞株SMMC-7721中抑制素的表达量较高。
