摘要
目的探讨旋毛虫肌幼虫虫体蛋白对小细胞肺癌H446细胞凋亡的影响及其作用的可能机制。方法将H446细胞与0.2mg/mL、0.4mg/mL、0.6mg/mL、0.8mg/mL、1.0mg/mL、1.2 mg/mL虫体蛋白分别培养并设为实验组,不处理的H446细胞设为对照组,采用噻唑兰(MTT)比色法检测旋毛虫肌幼虫虫体蛋白对H446细胞增殖活性的抑制作用;采用流式细胞术(FCM)检测旋毛虫肌幼虫虫体蛋白诱导H446细胞株凋亡的情况;采用Real-timePCR及Western blot法检测细胞色素C(cytochrome C,Cyt-C)和凋亡蛋白酶活化因子(apoptotic protease activating factor 1,Apaf-1)mRNA及蛋白的表达。结果 MTT比色法结果表明虫体蛋白能够抑制H446细胞增殖活性;流式细胞术结果表明虫体蛋白作用于H446细胞24h后,有明显的促凋亡作用;Real-timePCR及Western blot法结果显示,与阴性对照组相比,促凋亡基因Cyt-C和Apaf-1表达均上调。结论旋毛虫肌幼虫虫体蛋白能够抑制H446细胞增殖活性,并诱导细胞凋亡,其作用可能与Cyt-C和Apaf-1表达上调有关。
To probe the effects of Trichinella spiralis muscle larval somatic proteins on small cell lung cancer H446 cells and the possible mechanism of anti-tumor,H446 cells were culture with 0.2 mg/mL,0.4 mg/mL,0.6 mg/mL,0.8 mg/mL,1.0 mg/mL,and 1.2 mg/mL somatic proteins respectively.The experimental group was set and no dosing as control group.MTT colorimetric assay was used to test the effects of T.spiralis muscle larval somatic proteins on the proliferative activity of H446 cells.We used flow cytometry(FCM)to test the influence of T.spiralis muscle larval somatic proteins induced H446 cells apoptosis.The real-time PCR and Western blot methods were used to detect the expression of Cyt-C and apoptotic protease activating factor 1(Apaf-1)mRNA and protein.The MTT colorimetric assay showed that T.spiralis muscle larval somatic proteins could inhibit the proliferation of H446 cells;the flow cytometry showed that polypide proteins acted on H446 cells after 24 h appeared an obvious effect on promoting apoptosis.Results of real-time PCR and Western blot analysis indicated that compared with the control group,Cyt-C and Apaf-1 showed up-regulated expression.T.spiralis muscle larval somatic proteins could inhibit proliferation activity and induce the apoptosis of H446 cells,and its effects may be related to up-regulated expression of Cyt-C and Apaf-1.
作者
于莉
苏萌
谢广成
李丹
邴玉艳
方艳辉
赵蕾
杜娈英
YU Li;SU Men;XIE Guang-cheng;LI Dan;BING Yu-yan;FANG Yan-hui;ZHAO Lei;DU Luan-ying(Department of Pathogenic Biology,Chengde Medical University,Chengde 067000,China)
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2018年第2期124-128,共5页
Chinese Journal of Zoonoses
基金
河北省教育厅重点资助课题(No.ZD2014049)
河北省高等学校重点学科(No.201709)
承德医学院重点资助项目联合资助~~
关键词
旋毛虫
肌幼虫
虫体蛋白
小细胞肺癌
细胞凋亡
Trichinella spiralis
muscle larvae
somatic proteins
small cell lung cancer
apoptosis
