Journal of Hebei Medical University ›› 2023, Vol. 44 ›› Issue (7): 767-772.doi: 10.3969/j.issn.1007-3205.2023.07.005
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Abstract: Objective To explore the mechanism of zerumbone (Zer) in the treatment of mucoepidermoid carcinoma (MEC), in order to provide new ideas and theoretical basis for the development of MEC-related targeted drugs in clinical practice. Methods MEC cell lines H292 and H3118 were purchased, and the IC50 of Zer on MEC cell lines was detected by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Cell proliferation was detected by colony formation assay, and cell apoptosis and surface calreticulin(CRT) protein levels were detected by flow cytometry. The expression level of highmobility group protein B1(HMGB1) was detected by Western blot, to observe whether Zer could affect the immunogenic cell death(ICD) of MEC. Finally, to explore the possible molecular regulatory mechanism of Zer,the level of signal transducer and activator of transcription 3(STAT3) in MEC cells was up-regulated by pcDNA3.1-STAT3, and the transfection efficiency was detected by qRT-PCR. The expression levels of endoplasmic reticulum stress-related factors activating transcription factor 4(ATF4), ATF3 and C/EBP-homologousprotein(CHOP) were detected by real-time quantitative PCR (qRT-PCR) and Western blot. The adenosine triphosphate (ATP) level in the medium was detected by ATP assay kit. Results The apoptosis rate of cells in the Zer group was higher than that in the control group (P<0.05), and the HMGB1 protein level in the supernatant of H3118 cells in the Zer group was higher than that in the control group (P<0.05). The ATP level in H3118 cells in the Zer group was higher than that in the control group (P<0.05), and the protein and mRNA expression levels of endoplasmic reticulum stress-related genes ATF4, ATF3, and CHOP in the Zer group were higher than those in the control group (P<0.05). The STAT3 mRNA level in MECs in the Zer group was lower than that in the control group (P<0.05), and the protein and mRNA expression levels of endoplasmic reticulum stress-related genes ATF4, ATF3 and CHOP in the Zer+oe STAT3 group were lower than those in the Zer+oe NC group (P<0.05). Conclusion Zer-mediated STAT3 promotes endoplasmic reticulum stress in MEC cells and induces ICD of MEC cells.
Key words: carcinoma, mucoepidermoid, endoplasmic reticulum stress, immunogenic cell death
SU Feng-qin, SU Hui, WU Hui-wei, XING Shu-hua. Mechanism of zerumbone affecting mucoepidermoid carcinoma by immunogenic cell death [J]. Journal of Hebei Medical University, 2023, 44(7): 767-772.
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URL: https://xuebao.hebmu.edu.cn/EN/10.3969/j.issn.1007-3205.2023.07.005
https://xuebao.hebmu.edu.cn/EN/Y2023/V44/I7/767