蝎毒多肽提取物联合雷帕霉素增强H_(22)肝癌细胞自噬作用的机制研究  

Mechanism of Polypeptide Extract from Scorpion Venom Combined Rapamycin in Enhancing Autophagy of H_(22) Hepatoma Cells: an Experimental Study

作　　者：赵嵌嵌[1,2] 张维东[2] 武利存[2] 张璐璐[1,2] 王兆朋[2] 张月英[2] 王朝霞[2] 贾青[2] 

机构地区：[1]济南大学山东省医学科学院医学与生命科学学院,济南250062 [2]山东省医学科学院基础医学研究所病理室,济南250062

出　　处：《中国中西医结合杂志》2015年第7期866-870,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81073102;30873408);山东省自然科学基金资助项目(No.ZR2010HQ003)

摘　　要：目的观察蝎毒多肽提取物(polypeptide extract from scorpion venom,PESV)联合雷帕霉素(Rapamycin)对H22肝癌肿瘤细胞自噬的增强作用并探讨其作用机制。方法 40只昆明小鼠,采用皮下接种H22肝癌肿瘤细胞悬液法建立荷瘤模型,随机分为荷瘤对照组、PESV高剂量组、PESV低剂量组及联合组(PESV高剂量加雷帕霉素),每组10只。PESV高、低剂量组分别予20 mg/kg、10 mg/kg的PESV灌胃,联合组予雷帕霉素(2 mg/kg)及PESV(20 mg/kg)灌胃,连续给药干预14天,隔日测量肿瘤体积,记录肿瘤生长曲线,计算抑瘤率。HE染色观察各组肿瘤组织病理变化。免疫组织化学法检测各组肿瘤组织哺乳动物雷帕霉素靶蛋白(mammal target of rapamycin,m TOR)、UNC51样激酶1(UNC-51-like kinase-1,ULK1)、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,MAP1LC3A)及B淋巴细胞瘤蛋白质-2-相互作用蛋白质-1(Beclin1)蛋白表达水平。结果与荷瘤对照组比较,PESV高、低剂量组及联合组肿瘤生长受到明显抑制(P<0.05);与PESV高、低剂量组比较,联合组瘤重及体积明显减小(P<0.05),PESV高、低剂量组比较,差异无统计学意义(P>0.05)。免疫组织化学法显示,与荷瘤对照组比较,PESV高、低剂量组及联合组m TOR表达水平降低(P<0.05),ULK1、MAP1LC3A及Beclin1蛋白表达水平升高(P<0.05,P<0.01)。与PESV高剂量组比较,联合组ULK1、MAP1LC3A及Beclin1蛋白表达水平明显降低(P<0.05)。结论蝎毒多肽提取物联合化疗可能是通过抑制m TOR的活性,增强自噬相关蛋白ULK1、MAP1LC3A及Beclin1的表达,促进细胞自噬从而抑制肿瘤的发展。Objective To observe enhanced effects of polypeptide extract from scorpion venom（ PESV） combined Rapamycin on autophagy of H22 hepatoma cells in mice and to explore its possible mechanism. Methods The H22 hepatocarcinoma cell suspension was subcutaneously inoculated into 40 Kunming mice. Then tumor-bearing mice were randomly divided into four groups,i. e.,the control group,the high dose PESV group,the low dose PESV group,and the combination group（ high dose PESV ＋ Rapamycin）,10 in each group. Mice in high and dose PESV groups were administered with 20 mg / kg and 10 mg / kg PESV respectively by gastrogavage. Mice in the combination group were administered with 2 mg / kg rapamycin and 20 mg / kg PESV by gastrogavage. The intervention lasted for 14 successive days. The tumor volume was measured once every other day,the tumor growth curve was drawn,and then the tumor inhibitory rate calculated. Pathological changes of the tumor tissue were observed by HE staining. Protein expression levels of mammal target of rapamycin（ m TOR）,UNC-51-like kinase-1（ ULK1）,microtubule-associated protein1 light chain3（ MAP1LC3A）,and Beclin1 were detected by immunohistochemical assay. Results The growth of H22 hepatoma transplantation tumor was inhibited in high and low dose PESV groups and the combination group（P〈0. 05）. And there was statistical difference in tumor weight and tumor volume between the combination group and high and low dose PESV groups（P〈0. 05）. There was no statistical difference in tumor weight or tumor volume between the high dose PESV group and the low dose PESV group（P〈0. 05）. Immunohistochemical assay showed that the protein expression of m TOR was higher,but protein expressions of ULK1,MAP1LC3 A,Beclin1 were lower in the control group than in the rest 3 groups（P〈0. 05,P〈0. 01）. Compared with the high dose PESV group,protein expressions of ULK1,MAP1LC3 A,and Beclin1 were obviously lower（P〈0. 05）. Conclusion PESV combined Rapamycin might inhibit the development

关 键 词：自噬 蝎毒多肽提取物 雷帕霉素 H22肝癌细胞移植瘤 

分 类 号：R735.7[医药卫生—肿瘤]