皖南蝮蛇毒抑瘤组分I对人胃癌细胞SGC-7901荷瘤鼠体内抑制作用的研究

中国临床药理学与治疗学 ›› 2012, Vol. 17 ›› Issue (5): 491-496.

皖南蝮蛇毒抑瘤组分I对人胃癌细胞SGC-7901荷瘤鼠体内抑制作用的研究

卢林明¹, 周珏¹, 张根葆^2,3

¹皖南医学院病理教研室, ²病理生理教研室, ³蛇毒研究所,芜湖 241002,安徽

收稿日期:2012-02-12 修回日期:2012-04-15 出版日期:2012-05-26 发布日期:2012-05-28
通讯作者: 张根葆,男,教授,硕士生导师,主要从事心血管病理生理学、蛇毒应用与基础研究。E-mail: zgb858@163.com
作者简介:卢林明,男,硕士研究生,主要从事肿瘤病理学及抗肿瘤生物毒素的研究。Tel: 13855383399 E-mail: llm7172@sina.com
基金资助:
安徽省科技厅基金资助项目(10021303009);芜湖市科技局科研基金项目(09卫生类1-1)

In vivo tumor-inhibitory effects of Agkistrodon halys venom antitumor component I on tumor-bearing mice with human gastric cancer cell SGC-7901

LU Lin-ming¹, ZHOU Jue¹, ZHANG Gen-bao^2,3

¹Department of Pathology, ²Department of Pathophysiology, ³Institute of Snake Venom, Wannan Medical College, Wuhu 241002, Anhui, China

Received:2012-02-12 Revised:2012-04-15 Online:2012-05-26 Published:2012-05-28

摘要/Abstract

摘要： 目的:研究皖南蝮蛇毒抑瘤组分I(Agkistrodon halys venom antitumor component I,AHVAC-I)对人胃癌细胞SGC-7901荷瘤鼠的体内抑制作用及瘤细胞增殖活性变化,并初步探讨其作用机制。方法: 取生长状态良好的浓度为1×10⁷个/mL的SGC-7901人胃癌细胞悬液 0.1 mL 接种于裸小鼠右侧腋窝皮下,待其成瘤后随机分为模型组、紫杉醇阳性对照组、AHVAC-I高、中、低剂量组,隔日连续给药5次,定期观察肿瘤生长情况,测量肿瘤体积,绘制肿瘤生长曲线并计算抑瘤率,HE染色观察瘤体形态学变化,免疫组化SP法检测Ki-67增殖指数,原位末端标记技术(TUNEL)法检测细胞凋亡指数。结果: AHVAC-I在 1.0 mg/kg 剂量时对SGC-7901细胞有抑制作用,抑瘤率达 42.2%;在 3.0 mg/kg 剂量时有显著抑制作用,抑瘤率达 78.5%。实验组肿瘤内Ki-67阳性细胞数量下降,凋亡细胞明显增多。结论:AHVAC-I能明显抑制裸鼠体内SGC-7901人胃癌细胞的生长,其作用机制可能与诱导胃癌细胞发生凋亡及抑制肿瘤细胞增殖有关。

关键词: 皖南蝮蛇毒抑瘤组分I, 胃癌, 裸小鼠, 抑制, 增殖, 凋亡

Abstract: AIM: To investigate the in vivo tumor-inhibitory effects of snake venom antitumor component I(AHVAC-I) from Agkistrodon halys living in south Anhui province on the tumor-bearing mice with human gastric cancer cell SGC-7901 and activities of the cell proliferation as well as its mechanisms.METHODS: Cell suspension of 0.1 mL was obtained from the well-grown cell SGC-7901 by cell count of 1×10⁷/mL and incubated subcutaneously into the nude mice via right axillary fossa. After tumor formation, the mice were randomized into groups of model controls, positive paclitaxiel controls, AHVAC-I of low, medium and low dosage in which the animals were managed with AHVAC-I every other day for 5 consecutive times. The tumor growth and its sizes were observed and measured for mapping the growth curve and calculating the inhibitory rate. HE staining was performed to examine the morphologic change of tumors, and streptavidin-peroxidase (SP) immunohistochemical technique was used to detect the proliferation index of Ki-67 and TUNEL method was for examination of the apoptotic index.RESULTS:AHVAC-I at dose of 1.0 mg/kg produced inhibitory effects on SGC-7901 with inhibition rate of 42.2%, and inhibitory effects were significant at dose of 3.0 mg/kg with a rate of 78.5%. Ki-67 protein was reduced in the positive cells of treatment groups, leading to increased apoptosis of tumor cells.CONCLUSION: AHVAC-I can effectively inhibit the in vivo growth of gastric cell SGC-7901 in nude mice, and the possible mechanisms may be associated with the induced apoptosis and suppressed proliferation of the gastric tumor cells.

Key words: AHVAC-I, Gastric cancer, Nude mice, Inhibition, Proliferation, Apoptosis

中图分类号:

R965.2

卢林明, 周珏, 张根葆. 皖南蝮蛇毒抑瘤组分I对人胃癌细胞SGC-7901荷瘤鼠体内抑制作用的研究[J]. 中国临床药理学与治疗学, 2012, 17(5): 491-496.

LU Lin-ming, ZHOU Jue, ZHANG Gen-bao. In vivo tumor-inhibitory effects of Agkistrodon halys venom antitumor component I on tumor-bearing mice with human gastric cancer cell SGC-7901[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(5): 491-496.

参考文献

[1] 王国光,许敏,张根葆,等.蝮蛇毒提取物诱导白血病细胞K562凋亡[J].中国实验血液学杂志,2008,16(3):516-519.
[2] 张根葆,陈冬云,周志泳,等.蝮蛇毒蛋白C激活物的纯化与抗血小板活性[J].皖南医学院学报,2005,24(1):8-10.
[3] 张受涛,文尚武,周志泳,等.蝮蛇毒蛋白C激活物对凝血系统的影响[J].蛇志, 2006, 18(1):11-14.
[4] 许敏,张根葆.蝮蛇毒蛋白C激活物对大鼠心肌微血栓形成的抑制作用[J].皖南医学院学报,2008,27(4):239-242.
[5] 李晓红,蔡绍晖,任先达.蛇毒的抗肿瘤活性成分的研究进展[J].中国病理生理杂志,2004,20(10):1938-1941.
[6] Zhang L, Wu WT. Progress in studies on anticancer components from snake venom[J]. Chin J Zoology, 2003, 38(6): 120-124.
[7] Wei SX. Inhibitory effect of Guangxi cobra venom on novel human palatal salivary gland cell line of adenoid cystic card noma[J]. Chin J Mod Med, 2006, 16(21): 3231-3234.
[8] Yang S, Tsai C, Lu M, et al. Effects of cardiotoxin III on expression of genes and proteins related to G2/M arrest and apoptosis in K562 cells[J]. Mol Cell Biochem, 2006, 300(1/2): 185-190.
[9] 鲁珏,徐飞鹏.白眉蝮蛇蛇毒细胞毒素对胃癌细胞的杀伤作用及对细胞超微结构的影响[J].细胞与分子免疫学杂志,2009,25(4):335-337.
[10] 张及禄,文慧民,孙德军,等.蝮蛇毒小分子多肽的分离、纯化及其抗肿瘤作用研究[J].中国生化药物杂志,2009,30(4):217-225.
[11] 张闻多,丁文惠.细胞凋亡与凋亡诱导子[J].中国病理生理杂志,2006,9(15): 1858-1862.
[12] Yang SH, Chien CM, Chang LS, et al. Involvement of c-jun N-terminal kinase in G2/M arrest and caspase-mediated apoptosis induced by cardiotoxinIII (Naja naja atra) in K562 leukemia cells[J]. Toxieon, 2007, 49(7): 966-974.
[13] Wang CC, Chiang YM, Sung SC, et al. Plumbagin indueces cell cycle arrest and apoptosis through reactive oxygen species/c-Jun N-terminal kinase pathways in human melanoma A375.S2 cells[J]. Cancer Lett,2008, 259(1): 82-98.
[14] Park MH, Son DJ, Kwak DH, et al. Snake venom toxin inhibits cell growth induction of apoptosis in neuroblastoma cells [J]. Arch Pharm Res, 2009, 32(11): 1545-1554.
[15] 矫毓娟,刘江红,许贤豪.细胞凋亡的检测方法[J].中国神经免疫学和神经病学杂志, 2004,11(1):53-56.