蝎毒多肽提取物对卵巢癌SKOV_3细胞增殖抑制作用的实验研究

Inhibitory effect of polypeptide extracts from scorpion venom on proliferation of ovarian cancer SKOV_3 cells

目的:观察蝎毒多肽提取物(PESV)对卵巢癌SKOV3细胞的增殖抑制作用及机制。方法:采用噻唑蓝(MTT)方法检测PESV对卵巢癌SKOV3细胞的增殖抑制作用,流式细胞术观察PESV对细胞周期的影响,免疫组织化学方法(IHC)检测PESV干预后,Survivin,Capase-12及p27蛋白水平表达的变化,Westernblot检测survivin蛋白含量水平。结果:MTT结果显示,PESV在40 mg.L-1以上对卵巢癌SKOV3细胞增殖有抑制作用(浓度为40 mg.L-1时,增殖抑制率为34.66%),且随剂量加大作用增大,呈现剂量效应关系;流式细胞术检测细胞周期显示,PESV干预后,处于S期的细胞减少,处于G0/G1期的细胞增多;免疫组织化学检测显示:PESV干预后,SKOV3细胞p27表达上调,表达Survivin的肿瘤细胞减少,而表达Caspase-12的细胞显著增加。DDP干预后,仅发现Caspase-12的细胞表达增加。Western blot检测结果显示,PESV组survivin蛋白表达量下降。结论:PESV对卵巢癌SKOV3细胞增殖具有抑制作用,其机制是通过抑制细胞增殖周期进行及促进细胞凋亡。

Objective： To determine the effect of the polypeptide extracts from scorpion venom （PESV） on proliferation and apoptosis in the ovarian cancer cell line SKOV3 cells, and the possible mechanism. Methods： MTT assay was used to evaluate the effect of PESV on proliferation in SKOV3 cells, flow cytometry to test the effect on the cell cycle, immunocytochemistry to detect the changes in the expression of surviving, capase-12 and p27, and Western blotting to detect the protein content of survivin. Results： MTT assay showed that PESV inhibited proliferation of SKOV3 cells; the inhibition rate was 34.66% at 40 mg· L^-1, and the rate increased with concentration increasing. Flow cytometry showed that the S phase cells decreased while the G0/G1 phase cells increased after treatment with PESV. Immunocytochemistry showed that the cells expressing p27 increased, cells expressing survivin decreased, and cells expressing caspase-12 significantly increased. After DDP treatment, only the cells expressing caspase-12 increased. Western blotting showed that PESV reduced surviving protein expression. Conclusion： PESV can inhibit proliferation of the ovarian cancer SKOV3 cells; this may be regulated through retarding cell cycle and promoting apoptosis.