摘要
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 investigate the inhibitory effect of Trichinella spiralis polypide protein on H7402 cell line in vitro and to assess its anti-tumor effect against hepatocellular carcinoma （ HCC ）. Methods： The TrichineUa spiralis polypide protein was isolated from adult worms and newborn mixture. Trichinella spiralis polypide protein was prepared through homogenization and centrifugation, and its concentrations were determined by UV-visable spectrometers. H7402 cells treated with TrichineUa spiralis polypide protein （at 0. 035,0. 070, 0. 140 mg/ml） served as treatment group, and normal hepatic cell line HL-7702 treated with TrichineUa spiralis polypide protein served as control group. Cell proliferation was detected by MTT; the effect of Trichinella spiralis polypide protein on migration and invasion ability of H7402 cell line was assessed by scarification test and invasion test. Cell apoptosis was observed by TUNEL. Apoptosis and cell cycle were analyzed by FCM. Results： Trichinella spiralis polypide protein was successfully prepared. The protein at 0. 035,0. 070,0. 140 mg/ml inhibited H7402 cells proliferation by （22.40 ± 13.80） %, （29.45 ± 16.80 ） %, and （ 39.38 ± 17.80 ） %, respectively. Apoptosis was observed by TUNEL and FCM, with an apoptosis rate of （39.07 ± 0.90）% ; the cell cycle was blocked at S phase. Scarification test and invasion test suggested that TrichineUa spiralis polypide protein inhibited the migration and invasion of H7402 cell line, with the inhibition rate being （63.79 ± 13.71 ）%. Conclusion： Trichinella spiralis polypide protein can inhibit the proliferation, migration, invision of H7402 cell line, and it might be a potential anti-cancer agent.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
2007年第5期440-444,共5页
Chinese Journal of Cancer Biotherapy
基金
国家高技术研究发展(863)计划资助项目(No.2006AA02Z451)
Supported by the National High Technology Research and Development Program of China(No.2006AA02Z451)
关键词
旋毛虫
抗肿瘤
抗侵袭
抗增殖
Trichinella spiralis
antitumor
anti-invasion
antiproliferation
