论文部分内容阅读
目的观察转录因子C/EBPβ在人永生化正常肝细胞和肝癌细胞系中的表达差异性,探讨其与肝癌细胞内质网应激介导的细胞死亡的相关性。方法培养人永生化正常肝细胞系HHL5、HL7702和肝癌SMMC7721、Bel7402、HepG2、Hep3B细胞系;Hep3B细胞用衣霉素诱导内质网应激细胞模型;倒置光学显微镜下观察细胞形态;MTT法检测细胞生长抑制率;Hoechst 33258染色后荧光显微镜下检测细胞凋亡;RT-PCR和Western blotting技术分别检测mRNA和蛋白的表达水平。结果 C/EBPβ的mRNA和蛋白亚型C/EBPβ-1在4种人肝癌细胞系和永生化正常肝细胞系中均有表达,但C/EBPβ-3仅在人永生化正常肝细胞系中表达,在肝癌细胞系中不表达;衣霉素能够上调人肝癌Hep3B细胞C/EBPβ的mRNA和蛋白表达水平,明显上调C/EBPβ-3的表达水平;衣霉素能够诱导人肝癌Hep3B细胞内质网应激介导的细胞死亡,表现为细胞凋亡和类凋亡两种方式。结论 C/EBPβ的蛋白亚型C/EBPβ-3在人永生化正常肝细胞中表达而在肝癌细胞中不表达;C/EBPβ参与内质网应激介导的人肝癌细胞死亡。
Objective To observe the difference of transcription factor C / EBPβ in human immortalized normal hepatocytes and hepatocellular carcinoma cell lines, and to explore the relationship between transcription factor C / EBPβ and endoplasmic reticulum stress-induced cell death in hepatoma cells. Methods Human immortalized normal hepatocytes HHL5 and HL7702 and hepatocellular carcinoma SMMC7721, Bel7402, HepG2 and Hep3B cell lines were cultured. Fluorescein was used to induce endoplasmic reticulum stress cells in Hep3B cells. Cell morphology was observed under inverted light microscope. MTT assay The cell growth inhibition rate was measured by Hoechst 33258 staining. The mRNA and protein expression levels were detected by RT-PCR and Western blotting respectively. Results C / EBPβ mRNA and protein subtype C / EBPβ-1 were expressed in all four human hepatocellular carcinoma cell lines and immortalized normal liver cell lines, but C / EBPβ-3 was only expressed in human immortalized normal liver cell lines The expression of C / EBPβ-3 was significantly upregulated by tunicamycin in Hep3B cells. Tunicamycin could up-regulate the mRNA and protein expression of C / EBPβ in Hep3B cells, Cytoplasmic stress-mediated cell death, manifested as apoptosis and apoptosis in two ways. Conclusion The protein subtype C / EBPβ-3 of C / EBPβ is expressed in human immortalized normal liver cells but not in HCC cells. C / EBPβ is involved in endoplasmic reticulum stress-mediated death of human hepatoma cells.