蛇毒半胱氨酸蛋白酶抑制剂重组蛋白对人肝癌MHCC97H细胞TGF-β2、TNF-α表达的影响  

Effect of recombinant snake venom cystatin on TGF-β2 and TNF-α expression in MHCC97H cells

作　　者：谢群[1,2] 林扬元[2] 唐南洪[1,3] 林建银[1] 

机构地区：[1]福建医科大学分子医学研究中心消化道恶性肿瘤教育部重点实验室,福建福州350004 [2]福建省莆田学院基础医学院,福建莆田351100 [3]福建医科大学附属协和医院省肝胆外科研究所,福建福州350001

出　　处：《中国药理学通报》2013年第9期1208-1212,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 30371747);福建省科技重大项目(No 2002Y003);福建省教育厅资助省属高校科研专项(JK类)(No JK2011050)

摘　　要：目的研究蛇毒半胱氨酸蛋白酶抑制剂(snake venom cystatin,sv-cystatin)重组蛋白对人肝癌MHCC97H细胞转化生长因子-β2(transforming growth factor-β2,TGF-β2)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达的影响。方法采用ELISA方法检测不同浓度sv-cystatin重组蛋白对人肝癌MHCC97H细胞TGF-β2、TNF-α分泌的影响;实时定量PCR和Western blot法分别分析不同浓度sv-cystatin重组蛋白体外处理人肝癌MHCC97H细胞后TGF-β2、TNF-α的mRNA及蛋白表达。结果与对照组相比,5种浓度的sv-cystatin重组蛋白对MHCC97H细胞TGF-β2、TNF-α的分泌具有明显的抑制作用,并呈剂量依赖性(P<0.05);25、50、100、200 mg·L-14种浓度的重组蛋白能够抑制TGF-β2的mRNA及蛋白表达,与对照组比较差异具有统计学意义(P<0.05);而50、100、200 mg·L-13种浓度的重组蛋白能够抑制TNF-α的mRNA及蛋白表达,与对照组比较差异具有统计学意义(P<0.05)。结论 sv-cystatin重组蛋白可能通过抑制TGF-β2和TNF-α的表达来发挥多方面的抗肿瘤作用。Aim To investigate the effect of recombi- nant snake venome cystatin （sv-cystatin） on transfor- ming growth factor-β2 （TGF-β2） and tumor necrosis factor-α（TNF-α） expression in human hepatocellular carcinoma （MHCC97H） cells. Methods MHCC97H cells were treated with different concentrations of re- combinant sv-cystatin, then TGF-β2 and TNF-α secre- tion were assessed by ELISA. The mRNA or protein expression of TGF-β2 and TNF-α in MHCC97H cells were determined by real-time PCR or Western blot, re- spectively. Results Exposure of MHCC97H cells to increasing doses of recombinant sv-cystatin suppressed the secretion of TGF-βand TNF-α into the surround- ing medium in a dose-dependent manner compared to control （P 〈 0.05 ）. The mRNA and protein expression of TGF-β2 were significantly lower at 25,50, 100 and 200 mg· L-1 concentrations of recombinant sv- cystatin than those in the control groups （ P 〈 0. 05 ）. 50, 100 and 200 mg· L-1 concentrations of recombi- nant sv-cystatin down-regulated the mRNA and protein expression of TNF-α compared with those in control groups （P 〈 0. 05 ）. Conclusion Recombinant sv-cys- tatin can down-regulate the expression of TGF-β2 and TNF-α in MHCC97H cells, which contributes in part to the anti-tumor effect of sv-cystatin through multiple targets.

关 键 词：蛇毒 半胱氨酸蛋白酶抑制剂 肝癌 MHCC97H 转化生长因子Β2 肿瘤坏死因子α 

分 类 号：R73-3[医药卫生—肿瘤] R735.702.2[医药卫生—临床医学]