基于PI3K/Akt/FoxO4信号通路探究美洲大蠊提取物CⅡ-3诱导卵巢癌SKOV3细胞衰老的作用机制  

Exploring effect and mechanism of Periplaneta americana extract CII-3 to induce senescence of ovarian cancer SKOV3 cells based on the PI3K/Akt/FoxO4 signaling pathway

作　　者：田露 刘衡 张成桂 周玥 毕子莹 吴渟 TIAN Lu;LIU Heng;ZHANG Chenggui;ZHOU Yue;BI Ziying;WU Ting(Department of Histology and Embryology,Dali University,Dali 671000,China;Yunnan Provincial Key Laboratory of Aging and Disease Research,Dali 671000,China;Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical R&D,Dali 671000,China)

机构地区：[1]大理大学组织学与胚胎学教研室,云南大理671000 [2]云南省教育厅衰老与疾病研究重点实验室,云南大理671000 [3]云南省昆虫生物医药研发重点实验室,云南大理671000

出　　处：《现代药物与临床》2024年第9期2204-2211,共8页Drugs & Clinic

基　　金：国家自然科学基金资助项目(81860038,81660731)。

摘　　要：目的 探讨美洲大蠊提取物CⅡ-3对卵巢癌SKOV3细胞的抗肿瘤作用及其潜在作用机制。方法 体外培养SKOV3细胞,分为对照组和CⅡ-3组,通过CCK-8法和细胞集落形成实验测定SKOV3细胞增殖能力;流式细胞术检测SKOV3细胞线粒体膜电位和细胞周期的改变;衰老相关-β-半乳糖苷酶(SA-β-Gal)染色法检测SKOV3细胞衰老阳性细胞率;荧光定量PCR检测SKOV3细胞端粒酶逆转录酶(TERT)和磷脂酰肌醇-3-激酶(PI3K)、蛋白激酶B(Akt)、叉头盒蛋白O4(FoxO4)mRNA表达情况;Westernblotting检测p-PI3K、p-Akt、p-FoxO4蛋白表达情况。结果 与对照组比较,美洲大蠊提取物CⅡ-3可有效抑制SKOV3细胞增殖,细胞集落形成能力明显下降,细胞阻滞于G0/G1期,线粒体膜电位下降,SA-β-Gal细胞衰老阳性率增加,TERT活性降低,PI3K、Akt mRNA和p-PI3K、p-Akt蛋白表达上调,FoxO4 mRNA和p-FoxO4蛋白表达均下调。结论 美洲大蠊提取物CⅡ-3通过调控PI3K/Akt/FoxO4信号通路诱导SKOV3细胞衰老。Objective To investigate the anti-tumour effects and potential mechanisms of the Periplaneta americana extract C II-3 on ovarian cancer SKOV3 cells.Methods In vitro,SKOV3 cells were cultured and divided into control group and C II-3 group,and the proliferative capacity of SKOV3 cells was determined by the Cell Counting Kit-8(CCK-8)and the cell colony formation assay.Flow cytometry was used to detect the changes in the cell cycle and mitochondrial membrane potential of SKOV3 cells.Positive senescence-associated betagalactosidase(SA-β-Gal)staining assay to detect senescence in SKOV3 cells.q-PCR was utilized to assess the modification of telomerase reverse transcriptase(TERT),and PI3K,Akt,FoxO4 mRNA in SKOV3 cells.Western blotting assays were conducted to determine the expression of p-PI3K,p-Akt,p-FoxO4 protein in cells.Results Compared with the control group,Periplaneta americana extract CII-3 exhibited a significant inhibitory effect on the proliferation of SKOV3 cells,accompanied by a notable reduction in colony-forming ability,an increase in the proportion of cells arrested in the Go/Gi phase.The mitochondrial membrane potential decreased,the positive rate of senescence of SA-β-Gal cells increased,the activity of TERT decreased,the expression of PI3K,Akt mRNA and p-PI3K,p-Akt protein expression were up-regulated,and both FoxO4 mRNA and p-FoxO4 protein expression were down-regulated.Conclusion Periplaneta americana extract CII-3 induced cellular senescence in SKOV3 cells by regulating the PI3K/Akt/FoxO4 signaling pathway.

关 键 词：美洲大蠊 卵巢癌 细胞衰老 PI3K/Akt/FoxO4信号通路 磷脂酰肌醇-3-激酶 

分 类 号：R965[医药卫生—药理学]