The effect and mechanism of differentiation of HepG2 cells induced by mouse embryonic hepatocytes at day 13. 5 of gestation
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1. College of Integrated Chinese and Western Medicine of Hebei Medical University, Shijiazhuang 050017, China. 2. Hebei Key Lab of Laboratory Animal Science of Hebei Medical University, Shijiazhuang 050017. 3. Xingtai Medical College, Xingtai 054000

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R-33

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    Abstract:

    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.

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History
  • Received:June 15,2021
  • Revised:
  • Adopted:
  • Online: June 20,2022
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