摘要
目的在体外检测Nrf3基因对肝癌SMMC7721的生长影响作用。方法构建其融合蛋白真核表达载体,脂质体介导该真核表达载体转染肝癌SMMC7721细胞系,FCM观察其在体外对肝癌SMMC7721细胞系细胞周期和凋亡的影响;荧光显微镜观察候选基因亚细胞定位。结果Nrf3在肝癌SMMC7721细胞中表达,荧光显微镜下观察Nrf3定位于细胞核内。FCM分析显示Nrf3影响下肝癌SMMC7721G2/M+S期细胞所占比例较对照组明显减少,G0/G1细胞所占比例明显增加。证实Nrf3可抑制肝癌SMMC7721的DNA合成和有丝分裂,促使细胞阻滞于G0/G1期,抑制肝癌SMMC7721细胞的体外生长。FCM分析,未观察到明显的"亚G1"峰(即凋亡峰),提示Nrf3在体外对肝癌SMMC7721细胞的凋亡无影响。结论Nrf3在体外具有抑制肝癌细胞增殖的功能,对肝癌细胞的凋亡无影响,为肿瘤抑制基因。
Objective To observe the effect of Nfr3 on liver cancer SMMC7721 cell line in vitro.Methods Plasmids were constructed by inserting the candidate gene and transfected into liver cancer SMMC7721.Cell cycle and apoptosis of liver cancer SMMC7721 cell line in vitro were detected by flow cytometry（FCM）,and the subcellular localization of this candidate gene was observed under a fluorescence microscope.Results The expression vector pEGFP-N1-Nrf3 was constructed.FCM showed that the number of SMMC7721 cells in G2/M＋S and G0/G1 was significantly decreased and increased,respectively,under the influence of Nrf3.DNA synthesis and mitosis of SMMC7721 were restrained by Nrf3.FCM did not show the peak of deuto-G1（the peak of apoptosis）.Conclusion As a tumor-suppressing gene,Nrf3 can inhibit in vitro cell proliferation of Civer cancer cells but has no effect on their apoptosis.
出处
《军医进修学院学报》
CAS
2010年第9期924-926,共3页
Academic Journal of Pla Postgraduate Medical School
