小白菊内酯对宫颈癌细胞增殖、凋亡及PTEN/AKT通路的影响

Effects of parthenolide on proliferation,apoptosis and PTEN/AKT pathway of cervical cancer cells

目的研究小白菊内酯对宫颈癌细胞增殖、凋亡及磷酸酶及张力蛋白同系物/蛋白激酶(PTEN/AKT)通路的影响。方法体外培养宫颈癌Hela细胞,分别用不同浓度(10、50、60 mg/L)小白菊内酯(Par)处理对数期生长的Hela细胞为实验组,未经处理的Hela细胞为阴性对照组,2.5 mg/L顺铂处理为阳性对照组。采用四甲基偶氮唑蓝(MTT)法检测各组宫颈癌Hela细胞增殖情况;采用平板克隆试验法检测宫颈Hela细胞增殖能力;采用AnnexinV⁃FITC/碘化丙锭(PI)双染法体外检测宫颈癌Hela细胞凋亡;采用免疫印迹法(WB)检测宫颈癌Hela细胞中PTEN/AKT通路相关蛋白及凋亡相关蛋白表达。结果与阴性对照组比较,顺铂组、5、10、20、40、50 mg/L Par组对Hela细胞增殖抑制率均显著升高,差异有统计学意义(P均<0.05),且呈浓度依赖性和时间依赖性,其中50 mg/L Par作用48 h时,对Hela细胞抑制率达51.04%。与阴性对照组比较,顺铂组、10、50、60 mg/L Par组Hela细胞克隆形成率及B淋巴细胞瘤-2(Bcl⁃2)、p⁃AKT蛋白水平降低,细胞凋亡率及B淋巴细胞瘤2相关X蛋白(Bax)、cleaved⁃Caspase8、cleaved⁃Caspase3、PTEN蛋白水平升高,差异有统计学意义(P<0.05)。其中50、60 mg/L Par组与顺铂组Hela细胞克隆形成率、凋亡率及Bcl⁃2、p⁃AKT、Bax、cleaved⁃Caspase8、cleaved⁃Caspase3、PTEN蛋白水平比较,差异均无统计学意义(P>0.05)。结论小白菊内酯可能通过抑制PTEN/AKT通路,从而抑制宫颈癌Hela细胞增殖,促进其凋亡。

Objective To study the effects of parthenolide on proliferation,apoptosis and phosphatase and tensin homologue/protein kinase(PTEN/AKT)pathway of cervical cancer cells.Methods Hela cells of cervical cancer were cultured in vitro.The logarithmic Hela cells treated with different concentrations of Par(10,50,60 mg/L)were used as experimental group.The untreated Hela cells were treated as negative control group and 2.5 mg/L cisplatin treated Hela cells as positive control group.The proliferation of Hela cells was detected by MTT assay;the proliferative ability of Hela cells was detected by plate cloning test;Annexin V⁃FITC/PI double staining method was used to detect Hela cell apoptosis in cervical cancer in vitro,and the expressions of PTEN/AKT pathway⁃related proteins and apoptosis⁃related proteins in Hela cells of cervical cancer were detected by immunoblotting(WB).Results Compared with the negative control group,the inhibition rates of cisplatin group,5,10,20,40 and 50 mg/L Par groups were significantly increased(all P<0.05),and were concentration⁃dependent and time⁃dependent.The inhibition rate of 50 mg/L Par on Hela cell reached 51.04%at 48 h.Compared with negative control group,the clone formation rate and B⁃cell lymphoma⁃2(Bcl⁃2),p⁃AKT protein levels of Hela cells in cisplatin treated group,Par 10,50 and 60 mg/L groups decreased,while the apoptotic rate and Bax,cleaved⁃Caspase 8,cleaved⁃Caspase 3 and PTEN protein levels increased(P<0.05).There were no statistically significant differences in the cloning formation rate,apoptosis rate and Bcl⁃2,p⁃AKT,Bax,cleaved⁃Caspase8,cleaved⁃Caspase3 and PTEN protein levels between the 50,60 mg/L Par groups and cisplatin group(P>0.05).Conclusion Parthenolide might inhibit the proliferation and promote apoptosis of Hela cells by inhibiting PTEN/AKT pathway.