美洲大蠊提取物CⅡ-3调控p16^(INK4a)诱导SKOV3细胞衰老

Periplaneta americana extract CⅡ-3 inducing SKOV3 cells aging by regulating p16^(INK4a)protein

目的探讨美洲大蠊提取物CⅡ-3调控p16^(INK4a)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p16^(INK4a)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80μg/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G_(0)/G_(1)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p16^(INK4a)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p16^(INK4a)蛋白表达诱导SKOV3细胞衰老。

Objective To investigate the effect of Periplaneta americana extract CⅡ-3 in regulating p16^(INK4a)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 p16^(INK4a)protein expression.Results Periplaneta americana extract CⅡ-3 could effectively inhibit the proliferation of SKOV3 cells,and the optimal concentration was 80μg/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 G_(0)/G_(1)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 p16^(INK4a)protein were raised.Conclusion Periplaneta americana extract CⅡ-3 could induce senescence in SKOV3 cells by modulating protein expression of the senescence regulatory molecule p16^(INK4a).