目的 探討美洲大蠊提取物CII-3對(duì)卵巢癌SKOV3細(xì)胞的抗腫瘤作用及其潛在作用機(jī)制。方法 體外培養(yǎng)SKOV3細(xì)胞，分為對(duì)照組和CⅡ-3組，通過CCK-8法和細(xì)胞集落形成實(shí)驗(yàn)測(cè)定SKOV3細(xì)胞增殖能力；流式細(xì)胞術(shù)檢測(cè)SKOV3細(xì)胞線粒體膜電位和細(xì)胞周期的改變；衰老相關(guān)-β-半乳糖苷酶（SA-β-Gal）染色法檢測(cè)SKOV3細(xì)胞衰老陽性細(xì)胞率；熒光定量PCR檢測(cè)SKOV3細(xì)胞端粒酶逆轉(zhuǎn)錄酶（TERT）和磷脂酰肌醇-3-激酶（PI3K）、蛋白激酶B（Akt）、叉頭盒蛋白O4（FoxO4）mRNA表達(dá)情況；Western blotting檢測(cè)p-PI3K、p-Akt、p-FoxO4蛋白表達(dá)情況。結(jié)果 與對(duì)照組比較，美洲大蠊提取物CⅡ-3可有效抑制SKOV3細(xì)胞增殖，細(xì)胞集落形成能力明顯下降，細(xì)胞阻滯于G₀/G₁期，線粒體膜電位下降，SA-β-Gal細(xì)胞衰老陽性率增加，TERT活性降低，PI3K、Akt mRNA和p-PI3K、p-Akt蛋白表達(dá)上調(diào)，FoxO4 mRNA和p-FoxO4蛋白表達(dá)均下調(diào)。結(jié)論 美洲大蠊提取物CⅡ-3通過調(diào)控PI3K/Akt/FoxO4信號(hào)通路誘導(dǎo)SKOV3細(xì)胞衰老。

Objective To investigate the anti-tumour effects and potential mechanisms of the Periplaneta americana extract CⅡ-3 on ovarian cancer SKOV3 cells. Methods In vitro, SKOV3 cells were cultured and divided into control group and CⅡ-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 beta-galactosidase (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 G₀/G₁ 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.

R965

國(guó)家自然科學(xué)基金資助項(xiàng)目(81860038,81660731)