Overexpression of Cry2 Suppresses Proliferation and Induces Apoptosis in Hepatocellular  Carcinoma Cells by Mediating Ubiquitination and Degradation of c-Myc

doi:10.3870/j.issn.1672-0741.25.08.035

Abstract

Abstract: Objective To investigate the effect and underlying mechanism of cryptochrome 2(Cry2)gene overexpression on the proliferation and apoptosis of hepatocellular carcinoma(HCC)cells.Methods The expression levels of Cry2 mRNA and protein in human immortalized hepatocytes(THLE-3)and HCC cell lines(HepG2,HuH-7,SNU-398,and MHCC97-H)were determined using qRT-PCR and Western blot.Cry2 overexpression plasmid(OE-Cry2),c-Myc overexpression plasmid(OE-c-Myc),and their respective negative controls(Vector)were transfected into HuH-7 cells.Cells were treated with either 10 μmol/L proteasome inhibitor(MG132)or 100 μg/mL cycloheximide(CHX).The mRNA expression levels of Cry2 and c-Myc were assessed by qRT-PCR,while Western blot was used to evaluate the protein expression levels of Cry2,c-Myc,and cleaved Caspase-3.Cell proliferation was measured using CCK-8 assay,and the apoptosis rate was determined by flow cytometry.Additionally,the ubiquitination level of the c-Myc protein following Cry2 overexpression was analyzed using an immunoprecipitation(IP)assay.Results Compared with THLE-3 cells,the expression levels of Cry2 mRNA and protein in all HCC cell lines were significantly reduced(all P<0.05).Cry2 overexpression reduced the proliferation rate,induced apoptosis,upregulated cleaved Caspase-3 expression,and promoted ubiquitin-mediated degradation of c-Myc in HuH-7 cells.However,c-Myc overexpression reversed the effects of Cry2 overexpression on both cell proliferation and apoptosis in HuH-7 cells.Conclusion Overexpression of Cry2 may inhibit the proliferation and induce apoptosis of HCC cells.Its mechanism of action may be related to mediating c-Myc ubiquitination and degradation.

Key words: hepatocellular carcinoma, proliferation, apoptosis, cryptochrome 2, myelocytomatosis oncogene, ubiquitination

CLC Number:

R735.7

Xiong Jie, Liu Yang, Zhang Xihua, et al. Overexpression of Cry2 Suppresses Proliferation and Induces Apoptosis in Hepatocellular Carcinoma Cells by Mediating Ubiquitination and Degradation of c-Myc[J]. Acta Medicinae Universitatis Scientiae et Technologiae Huazhong, 2026, 55(4): 519-525.

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Overexpression of Cry2 Suppresses Proliferation and Induces Apoptosis in Hepatocellular Carcinoma Cells by Mediating Ubiquitination and Degradation of c-Myc

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