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目的:研究旋毛虫虫体蛋白对肝癌H7402细胞的抑制作用,确认其对肝癌细胞的抗肿瘤活性。方法:通过体内、体外培养获得旋毛虫成虫与新生幼虫的混合物,采用匀浆、离心方法制备旋毛虫虫体蛋白,紫外分光光度计检测蛋白浓度。分别设0.035、0.070、0.140 mg/ml与H7402细胞作用为实验组,并设虫体蛋白对正常肝细胞HL-7702细胞作用为对照组,采用MTT检测细胞增殖能力,划痕实验、侵袭实验检测旋毛虫虫体蛋白对H7402的迁移及侵袭能力的影响,采用TUNEL法和流式细胞术分析肿瘤细胞凋亡和细胞周期。结果:成功制备了旋毛虫虫体蛋白。0.035、0.070、0.140 mg/ml旋毛虫虫体蛋白对H7402细胞增殖的抑制率分别为(22.40±13.80)%、(29.45±16.80)%、(39.38±17.80)%。TUNEL实验和流式细胞术可观察到旋毛虫虫体蛋白诱导H7402凋亡,其凋亡率为(39.07±0.90)%,细胞周期阻滞于S期。划痕和侵袭的实验表明旋毛虫虫体蛋白对H7402细胞迁徙能力有抑制作用,对其侵袭的抑制率为(63.79±13.71)%。结论:旋毛虫虫体蛋白对肝癌H7402细胞的增殖、迁移和侵袭具有抑制作用,可能是一个有潜在应用价值的抗肝癌生物制剂。
Objective: To study the inhibitory effect of Trichinella spiralis protein on hepatocarcinoma H7402 cells and to confirm its anti-tumor activity on hepatoma cells. Methods: The mixture of adult Trichinella spiralis and neonatal larva was obtained in vitro and in vivo. The protein of Trichinella spiralis was prepared by homogenization and centrifugation. The protein concentration was determined by ultraviolet spectrophotometer. The effects of 0.035,0.070,0.140 mg / ml and H7402 cells on the proliferation of HL-7702 cells were evaluated by MTT. The cell proliferation was evaluated by scratch assay and invasion assay Trichinella spiralis worms body protein H7402 migration and invasion ability, using TUNEL method and flow cytometry analysis of tumor cell apoptosis and cell cycle. Results: Trichinella spiralis protein was successfully prepared. The inhibitory rates of 0.035, 0.070,0.140 mg / ml Trichinella spiralis protein to H7402 cells were (22.40 ± 13.80)%, (29.45 ± 16.80)% and (39.38 ± 17.80)%, respectively. TUNEL assay and flow cytometry showed that the Trichinella spiralis protein induced the apoptosis of H7402 cells. The apoptotic rate was (39.07 ± 0.90)%. The cell cycle arrest was in S phase. Scratches and invasion experiments showed that Trichinella spiralis protein had inhibitory effect on migration of H7402 cells, and the inhibitory rate of Trichinella spiralis invasion was (63.79 ± 13.71)%. Conclusion: Trichinella spiralis protein has inhibitory effect on the proliferation, migration and invasion of H7402 cells, which may be a potentially useful anti-HCC biological agent.