• 首页
  • 期刊介绍
  • 编委会
  • 投稿指南
  • 期刊订阅
  • 广告合作
  • 留言板
  • 联系我们
  • English
肖志刚,郑 丽,王梓瀛,刘 昆,王 瑜,齐锦生,栗彦宁.小鼠第 13. 5 天胚肝细胞诱导 HepG2 细胞分化的作用机制[J].中国比较医学杂志,2022,32(4):47~53.
小鼠第 13. 5 天胚肝细胞诱导 HepG2 细胞分化的作用机制
The effect and mechanism of differentiation of HepG2 cells induced by mouse embryonic hepatocytes at day 13. 5 of gestation
投稿时间:2021-06-15  
DOI:10. 3969 / j.issn.1671-7856. 2022. 04. 007
中文关键词:  HepG2  细胞分化  胚肝细胞  β-Catenin  HNF-4α
英文关键词:HepG2  cell differentiation  embryonic hepatocytes  β-Catenin  HNF-4α
基金项目:
作者单位E-mail
肖志刚 河北医科大学中西医结合学院,石家庄 050017 1345449822@ qq.com 
郑 丽 1.河北医科大学中西医结合学院,石家庄 050017
3.邢台医学高等专科学校,河北 邢台 054000 
 
王梓瀛 河北医科大学中西医结合学院,石家庄 050017  
刘 昆 河北医科大学中西医结合学院,石家庄 050017  
王 瑜 河北医科大学实验动物学部,石家庄 050017  
齐锦生 河北医科大学中西医结合学院,石家庄 050017 qijinsheng777@ 163.com 
栗彦宁 河北医科大学实验动物学部,石家庄 050017  
摘要点击次数: 54
全文下载次数: 25
中文摘要:
       目的 探讨小鼠第 13. 5 天胚肝细胞是否通过抑制 β-Catenin 诱导人肝细胞肝癌细胞株 HepG2 细胞分化及其作用机制。 方法 用免疫荧光法检测肝细胞标记分子-白蛋白(ALB)的分布,以鉴定小鼠第 13. 5 天胚肝细胞。 小鼠第 13. 5 天胚肝细胞与 HepG2 细胞株共培养 24 h、48 h 后,采用 qRT-PCR方法检测 HepG2 细胞株中肝癌标记分子-甲胎蛋白(AFP)、肝细胞分化调控因子-肝细胞核因子 4α(HNF-4α)和成熟肝细胞标记分子-细胞色素 P450 家族成员 1B1(CYP1B1)和乙醇脱氢酶 1C(ADH1C)的 mRNA 表达。 共培养 48 h 后,观察 HepG2 细胞株形态变化,采用 Western blot 法检测 AFP、HNF-4α 和 β-Catenin 的蛋白含量,免疫荧光法检测 β-Catenin 蛋白在细胞的分布。 应用 β-Catenin 抑制剂处理后,观察 HepG2 细胞株形态变化并且检测 AFP 蛋白表达。 结果 原代培养细胞 ALB 荧光显示阳性。 与对照组相比,共培养后 HepG2 细胞株中,AFP 相对 mRNA 表达量在 24 h 和 48 h 均显著降 低,HNF-4α、CYP1B1 和 ADH1C 的相对 mRNA 表达量在 24 h 和 48 h 均显著升高。 与对照组相比,共培养 48 h 后, HepG2 细胞株生长明显抑制,细胞形态趋于正常肝细胞的六边形,AFP 和 HNF-4α 蛋白表达明显升高。 与对照组相比,共培养 48 h 后,β-Catenin 的蛋白含量明显降低,同时 HepG2 细胞株中 β-Catenin 的荧光明显减弱。 与对照组相比,β-Catenin 抑制剂处理可明显改变 HepG2 细胞株形态,引起共培养组 HepG2 细胞株形态更剧烈的变化,显著抑制 AFP 的蛋白表达。 结论 小鼠第 13. 5 天胚肝细胞可能主要通过抑制 β-Catenin,诱导 HepG2 细胞株分化。
英文摘要:
       Objective To explore the differentiation of HepG2 cells induced by mouse embryonic hepatocytes at day 13. 5 of gestation through the inhibition of β-Catenin. Methods The distribution of hepatocyte marker molecule- albumin (ALB) was detected by immunofluorescence to identify the hepatocytes of mouse embryo on day 13. 5. After HepG2 cells were co-cultured with embryonic hepatocytes at day 13. 5 of gestation for 24 h and 48 h, the mRNA expressions of AFP (biomarker of hepatocellular carcinoma), HNF-4α (controller of hepatocyte differentiation), CYP1B1 (biomarker of mature hepatocytes) and ADH1C ( another biomarker of mature hepatocytes) were measured by reverse transcriptional quantitative polymerase chain reaction (RT-qPCR). After HepG2 cells were co-cultured with day 13. 5 of gestation embryonic hepatocytes for 48 h, the morphology was observed; AFP, HNF-4α and β-catenin expressions were measured by Western blot and β-Catenin distribution was determined by immunofluorescence. Following treatment with the β-catenin inhibitor, morphology was observed and AFP content was measured. Results Most cells were ALB-positive. The relative mRNA expression of AFP decreased significantly after 24 h and 48 h co-culture, and HNF-4α, CYP1B1 and ADH1C increased significantly at 24 h and 48 h co-culture compared with the levels in the controls. The proliferation of HepG2 cells was suppressed and cell morphology tended to be hexagonal like normal hepatocytes; AFP protein content decreased and HNF-4α protein content increased after co-culture with embryonic hepatocytes at day 13. 5 of gestation for 48 h. Compared with the levels in the control, β-Catenin protein content decreased in the co-culture group and β-catenin was markedly attenuated in the co-cultured HepG2 cells in immunofluorescence result. Compared with the effects in the control group, β- catenin inhibitor treatment caused notable morphological changes of HepG2 cells and induced more dramatic morphological changes of HepG2 cells; furthermore, it reduced AFP protein content. Conclusions HepG2 cells may be induced to undergo differentiation by embryonic hepatocytes at day 13. 5 of gestation mainly through the suppression of β-catenin.
查看全文  查看/发表评论  下载PDF阅读器
关闭
您是第 5790124 位访问者
版权所有:中国实验动物学会 主管单位:中国科学技术协会 主办单位:中国实验动物学会 中国医学科学院医学实验动物研究所
地  址: 北京市朝阳区潘家园南里5号 邮编:100021 电话:010-67779337 E-mail:bjb@cnilas.org
本系统由北京勤云科技发展有限公司设计
微信关注二维码