灰树花多糖对宫颈癌细胞增殖和转移的影响及机制

Effects and mechanism of Grifola frondosa polysaccharide on the proliferation and metastasis of cervical cancer cells

目的探讨灰树花多糖对宫颈癌Caski细胞增殖、凋亡、侵袭迁移及PI3K/AKT(磷脂酰肌醇3⁃激酶/蛋白激酶B)信号通路的影响。方法50、100、200μg/mL灰树花多糖处理Caski细胞48 h,通过MTT比色法、Transwell小室及Annexin V⁃FITC/PI双染法分别检测细胞增殖、侵袭迁移及凋亡率,Western blot检测AKT、p⁃AKT、PCNA(增殖细胞核抗原)、Cleaved caspase⁃3(裂解的半胱天冬酶3)和E⁃cadherin蛋白(上皮钙黏蛋白)表达。使用PI3K/AKT信号通路抑制剂LY294002(10μmol/L)单独或联合灰树花多糖(10μmol/L LY294002和200μg/mL灰树花多糖)处理Caski细胞48 h,检测各组细胞增殖、侵袭迁移、凋亡及AKT、p⁃AKT、PCNA、Cleaved caspase⁃3和E⁃cadherin蛋白表达。结果随着灰树花多糖质量浓度增加细胞增殖、侵袭和迁移能力降低,凋亡率升高,p⁃AKT和PCNA表达降低,Cleaved caspase⁃3和E⁃cadherin表达升高(P<0.05)。LY294002可增强灰树花多糖对Caski细胞增殖、侵袭迁移、凋亡及p⁃AKT、PCNA、Cleaved caspase⁃3和E⁃cadherin表达的影响。结论灰树花多糖可通过阻断PI3K/AKT信号通路抑制宫颈癌Caski细胞增殖、侵袭和迁移,促进细胞凋亡。

AIM To investigate the effects of Grifola frondosa polysaccharide on the proliferation,apoptosis,invasion and migration of cervical cancer Caski cells and the PI3K/AKT signaling pathway.METHODS Caski cells were treated with 50,100 and 200μg/mL Grifola frondosa polysaccharides for 48 hours.Cell proliferation,invasion,migration and apoptosis were detected by MTT colorimetry,Transwell cell and annexin V⁃FITC/PI double staining.Expression of AKT,p⁃AKT,PCNA,cleaved caspase3 and E⁃cadherin were detected by Western blot.Caski cells were treated with PI3K/AKT signal pathway inhibitor LY294002(10μmol/L)alone or in combination with Grifola frondosa polysaccharide(10μmol/L LY294002 and 200μg/ml Grifola frondosa polysaccharide)for 48 hours,Cell proliferation,invasion,migration,apoptosis and expression of AKT,p⁃AKT,PCNA,cleaved caspase 3 and E⁃cadherin were detected.RESULTS With the increase of polysaccharide concentration in Grifola frondosa,the ability of cell proliferation,invasion and migration decreased,the apoptosis rate increased,the expression of p⁃AKT and PCNA decreased,the expression of cleaved caspase 3 and E⁃cadherin increased(P<0.05).LY294002 can enhance the effects of Grifola frondosa polysaccharide on the proliferation,invasion,migration,apoptosis and expression of p⁃AKT,PCNA,cleaved caspase 3 and E⁃cadherin of Caski cells.CONCLUSION Grifola frondosa polysaccharide can inhibit the proliferation,invasion and migration of Caski cells and promote apoptosis by blocking the PI3K/AKT signal pathway.