口腔医学 ›› 2023, Vol. 43 ›› Issue (10): 889-893.doi: 10.13591/j.cnki.kqyx.2023.10.005

• 基础研究 • 上一篇    下一篇

盐酸小檗碱通过调节自噬抑制口腔鳞状细胞癌增殖的研究

肖金枝,余伟(),张昊   

  1. 十堰市人民医院(湖北医药学院附属人民医院)口腔科,湖北十堰(442000)
  • 修回日期:2023-06-16 出版日期:2023-10-28 发布日期:2023-10-20
  • 通讯作者: 余伟 E-mail:yuweikouqiang@163.com
  • 基金资助:
    湖北省教育厅科研项目(B2022133);十堰市科技局项目(21Y51)

Berberine hydrochloride inhibits the proliferation of oral squamous cell carcinoma by regulating autophagy

XIAO Jinzhi,YU Wei(),ZHANG Hao   

  1. Department of Stomatology, Shiyan People’s Hospital(People’s Hospital Affiliated to Hubei Medical University), Shiyan 442000, China
  • Revised:2023-06-16 Online:2023-10-28 Published:2023-10-20

摘要:

目的 探讨盐酸小檗碱(berberine hydrochloride,BH)通过调节细胞自噬对抑制口腔鳞状细胞癌增殖的影响。方法 不同浓度的盐酸小檗碱(60、90、120、240 μmol/L)作用于口腔鳞癌细胞Cal-27 24、48、72 h后,CCK-8实验检测其对Cal-27细胞存活率的作用;单克隆实验(colony formation)检测其对Cal-27细胞增殖能力的影响;蛋白免疫印迹(Western Blot)检测盐酸小檗碱对Cal-27细胞增殖、自噬蛋白表达的作用。设置分组:对照组(0),单独药物组(90 μmol/L BH),雷帕霉素组(50 nmol/L RAPA)以及盐酸小檗碱+雷帕霉素共同处理组(90 μmol/L BH+50nmol/L RAPA)。以此分组进行CCK8实验、单克隆实验以及蛋白免疫印迹实验检测盐酸小檗碱抑制细胞自噬与抑制Cal-27细胞增殖的关系。结果 CCK-8结果示,盐酸小檗碱作用于Cal-27细胞后其存活率逐渐降低;克隆形成实验结果示,经盐酸小檗碱处理后,Cal-27细胞增殖能力明显削弱;Western Blot结果示,随着盐酸小檗碱浓度的增加,增殖蛋白PCNA的表达量逐渐降低,且自噬家族蛋白Beclin-1及LC3Ⅱ/Ⅰ的表达也逐渐降低。自噬激活剂(雷帕霉素)与盐酸小檗碱共同处理组,与单独药物组对比,其细胞活性及克隆形成能力增强,PCNA、Beclin-1及LC3Ⅱ/Ⅰ的表达上调。结论 盐酸小檗碱抑制口腔鳞状细胞癌Cal-27细胞的自噬反应,并通过抑制自噬进一步抑制其增殖。

关键词: 盐酸小檗碱, 口腔鳞状细胞癌, 自噬, 增殖

Abstract:

Objective To investigate the effect of berberine hydrochloride(BH)on inhibiting the proliferation of oral squamous cell carcinoma by regulating autophagy. Methods After different concentrations of BH (60, 90, 120, 240 μmol/L)acted on oral squamous cell carcinoma Cal-27 for 24 h, 48 h, 72 h, CCK-8 experiments detected the effect on Cal-27 cell viability. Colony formation detected the effect on the proliferation capacity of Cal-27 cell. Western Blot detected the effect of berberine hydrochloride on the expression of proliferation and autophagy protein of Cal-27 cell. The groups were set up: Control group (0), BH group (90 μmol/L BH), rapamycin (RAPA) group (50 nmol/L RAPA), and BH + RAPA cotreatment group (90 μmol/L BH+50 nmol/L RAPA). CCK8 experiments, colony formation experiments and Western Blot experiments were carried out in this group to detect the relationship between berberine hydrochloride inhibition of autophagy and inhibition of Cal-27 cell proliferation. Results CCK-8 results showed that the survival rate of berberine hydrochloride gradually decreased after it was applied to Cal-27 cells. The results of colony formation indicated that Cal-27 cells were significantly weakened after berberine hydrochloride treatment. The results of Western Blot demonstrated that with the increase of BH concentration, the expression of PCNA protein, which was a proliferation index, gradually decreased, and the expression of autophagy family protein Beclin-1 and LC3Ⅱ/Ⅰ also gradually decreased. Compared with the BH group, the cell activity and clonal formation ability of BH+RAPA group were enhanced, and the expression of PCNA protein, autophagy protein Beclin-1 and LC3Ⅱ/Ⅰ was upregulated. Conclusion BH inhibits the autophagy response of oral squamous cell carcinoma Cal-27 cell and further inhibits its proliferation by inhibiting autophagy.

Key words: berberine hydrochloride, oral squamous cell carcinoma, autophagy, proliferation

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