医学研究与教育 ›› 2024, Vol. 41 ›› Issue (1): 1-8.DOI: 10.3969/j.issn.1674-490X.2024.01.001

• 基础医学 •    下一篇

美洲大蠊提取物CⅡ-3调控p16INK4a诱导SKOV3细胞衰老

毕子莹1,周玥1,张成桂2,刘衡2,汪小波1,田露1,吴渟1   

  1. 1.大理大学基础医学院, 云南 大理 671000;
    2.云南省昆虫生物医药研发重点实验室, 云南 大理 671000
  • 收稿日期:2023-11-23 出版日期:2024-02-25 发布日期:2024-02-25
  • 通讯作者: 周玥(1979—),女,云南大理人,教授,博士,硕士生导师,主要从事中药药理学研究。E-mail: zhouyue_120@sina.com
  • 作者简介:毕子莹(2000—),女,辽宁铁岭人,在读硕士,主要从事中药药理学研究。 E-mail: 2836766736@qq.com
  • 基金资助:
    国家自然科学基金面上项目(81860038)

  • Received:2023-11-23 Online:2024-02-25 Published:2024-02-25

摘要: 目的 探讨美洲大蠊提取物CⅡ-3调控p16INK4a蛋白诱导卵巢癌细胞SKOV3衰老的作用。方法 通过CCK-8法测定不同浓度和时间美洲大蠊提取物CⅡ-3对SKOV3细胞增殖的作用;运用细胞集落形成实验测定美洲大蠊提取物CⅡ-3对SKOV3细胞增殖能力的影响;流式细胞术检测CⅡ-3作用后SKOV3细胞周期的变化;衰老相关-β-半乳糖苷酶(SA-β-Gal)染色检测CⅡ-3作用后SKOV3细胞衰老变化;Western Blotting检测CⅡ-3作用后SKOV3细胞衰老调控分子p16INK4a蛋白表达的变化。结果 美洲大蠊提取物CⅡ-3可有效抑制SKOV3细胞增殖,最佳作用浓度为80 μg/mL,作用时间为48 h;CⅡ-3作用后,SKOV3细胞集落形成能力下降;阻滞于G0/G1期细胞比例增高,S期细胞比例减少;SA-β-Gal阳性细胞百分比增加;衰老调控分子p16INK4a蛋白表达上调。结论 美洲大蠊提取物CⅡ-3通过调控衰老调控分子p16INK4a蛋白表达诱导SKOV3细胞衰老。

关键词: 美洲大蠊提取物CⅡ-3, SKOV3细胞, 衰老, p16INK4a蛋白

Abstract: Objective To investigate the effect of Periplaneta americana extract CⅡ-3 in regulating p16INK4a protein in inducing SKOV3 aging. Methods SKOV3 cells were treated with different concentrations of Periplaneta americana extract CⅡ-3,and the effects of CⅡ-3 on the proliferation of SKOV3 cells were detected by CCK-8 assay. The effect of Periplaneta americana extract CⅡ-3 on the colony formation of SKOV3 cells was determined by colony culture experiments. SKOV3 cell cycle changes after CⅡ-3 action were examined with flow cytometry detection. Senescence changes in SKOV3 cells after CⅡ-3 action were determined with SA-β-Gal staining detects. Western Blotting was used to detect the effects of CⅡ-3 on SKOV3 cells phosphorylated p16INK4a protein expression. Results Periplaneta americana extract CⅡ-3 could effectively inhibit the proliferation of SKOV3 cells, and the optimal concentration was 80 μg/mL and an action time was 48 h. The colony formation ratio of SKOV3 cells were reduced after the effect of Periplaneta americana extract CⅡ-3 on SKOV3 cells, the proportion of cells blocked in G0/G1 phase were increased, the proportion of cells in S phase decreased, the percentage of positive cells of SA-β-Gal was increased and the upregulation of the expression of the aging regulatory molecule p16INK4a protein were raised. Conclusion Periplaneta americana extract CⅡ-3 could induce senescence in SKOV3 cells by modulating protein expression of the senescence regulatory molecule p16INK4a.

Key words: Periplaneta americana extract CⅡ-3, SKOV3 cells, senescence, p16INK4a protein

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