眼镜蛇毒细胞毒素缓释微球瘤内注射对裸鼠人肝癌皮下移植瘤的影响

doi:10.3736/jcim20090908

Abstract

Objective

To evaluate the safety and efficacy of intratumoral injection of polylactic-co-glycolic acid (PLGA) microspheres containing cobra venom cytotoxin in nude mice with transplanted human hepatoma.
Methods

Cytotoxic activity of cytotoxin from cobra venom was determined by using methyl thiazolyl tetrazolium method in vitro. Microspheres containing cobra venom cytotoxin were prepared with a double emulsion-solvent evaporation method. Forty BALB/c nude mice were inoculated subcutaneously in right flank with hepatoma BEL-7404 cells. Thirty-two mice whose tumor size reached about 1.0 cm in diameter, were randomly assigned into normal saline group, blank microsphers group, cytotoxin group and cytotoxin-PLGA group. Nude mice were intratumorally injected with normal saline, blank microspheres, cytotoxin or cytotoxin-PLGA microspheres respectively. Internal echo characteristics and blood flow of tumors were observed by high-frequency ultrasound every week after treatment. Twenty-six days after treatment, the tumors were removed to calculate the inhibition rate of tumor growth. The tumor, heart, liver and kidney tissues were obtained for histopathological examination.
Results

The cytotoxin separated and purified from crude cobra venom caused intense cytotoxic effects to the BEL-7404 cells in vitro. The diameter of PLGA microspheres containing cobra venom cytotoxin was about (34.45±9.85) μm. Encapsulation rate was up to (78.13±8.92)%, and cumulative amount of cobra venom cytotoxin released from the PLGA microspheres in vitro during 30 days was up to 84.3%. After intratumoral injection, tumor volumes and weights in the cytotoxin-PLGA group were lower than those in the normal saline group, with a tumor growth inhibition rate of 52.36%. Observed under a light microscope, most tumor tissues were necrotic. No obvious morphological change could be seen on the liver, kidney and heart tissues.
Conclusion: The above findings indicate that intratumoral injection of cytotoxin-PLGA microspheres has strong antitumor effect and can obviously lessen systemic toxicity, which may provide an effective and feasible method for hepatocellular carcinoma treatment.

Key words: Cobra venom cytotoxin, Liver neoplasms, Drug therapy, Delayed-action preparation, Mice, Nude

Yan Wang , Li-wu Lin, Zhi-kui Chen, En-sheng Xue, Xiao-dong Lin, Li-yun Yu, Zhen-hu Lin. Effects of intratumoral injection of microspheres containing cobra venom cytotoxin on transplanted human hepatoma in nude mice. Journal of Chinese Integrative Medicine, 2009, 7(9): 836-841.

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URL: http://www.jcimjournal.com/EN/10.3736/jcim20090908

http://www.jcimjournal.com/EN/Y2009/V7/I9/836

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Figures/Tables 7

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References 14

[1]	Abu-Sinna G, Esmat AY, Al-Zahaby AA, Soliman NA, Ibrahim TM . Fractionation and characterization of Cerastes cerastes cerastes snake venom and the antitumor action of its lethal and non-lethal fractions[J]. Toxicon, 2003,42(2):207-215 doi: 10.1016/S0041-0101(03)00138-7
[2]	Feofanov AV, Sharonov GV, Dubinnyi MA, Astapova MV, Kudelina IA, Dubovskii PV, Rodionov DI, Utkin YN, Arseniev AS . Comparative study of structure and activity of cytotoxins from venom of the cobras Naja oxiana, Naja kaouthia, and Naja haje[J]. Biochemistry (Mosc), 2004,69(10):1148-1157 doi: 10.1023/B:BIRY.0000046890.46901.7e
[3]	Oron U, Chaim-Matyas A, Ovadia M . Histopathological changes in WEHI-3B leukemia cells following intoxication by cytotoxin P4 from Naja nigricollis nigricollis venom[J]. Toxicon, 1992,30(9):1122-1126 doi: 10.1016/0041-0101(92)90058-D
[4]	Chen ZK, Lin LW, Lin Q, Zhuang QQ, He YM, Xue ES . Preparation and characterization of poly(lactide-co-glycolide acid) microspheres containing cytotoxin from Naja Naja Atra venom[J]. Zhongguo Sheng Hua Yao Wu Za Zhi, 2008,29(3):161-164
	陈志奎, 林礼务, 林琦, 庄权权, 何以敉, 薛恩生 . 眼镜蛇毒细胞毒素缓释微球制备及体外性质研究[J]. 中国生化药物杂志, 2008,29(3):161-164
[5]	Sapin A, Garcion E, Clavreul A, Laqarce F, Benoit JP, Menei P . Development of new polymer-based particulate systerms for anti-glioma vaccination[J]. Int J Pharm, 2006,309(1-2):1-5 doi: 10.1016/j.ijpharm.2005.10.046 pmid: 16386390
[6]	DeFail AJ, Chu CR, Izzo N, Marra KG . Controlled release of bioactive TGF-beta 1 from microspheres embedded within biodegradable hydrogels[J]. Biomaterials, 2006,27(8):1579-1585 doi: 10.1016/j.biomaterials.2005.08.013
[7]	Sun L, Wang Y, Liu ML, Xu MD . Recent developments on PLA/PLGA microspheres and its application[J]. Yao Wu Fen Xi Za Zhi, 2005,25(11):1409-1415
	孙磊, 王岩, 刘明理, 许鸣镝 . 聚乳酸/乳酸-乙醇酸共聚物微球制剂研究的新进展[J]. 药物分析杂志, 2005,25(11):1409-1415
[8]	Huang WD, Long DH, Gu HG . Explore of preparation and formulation parameters on BSA-PLGA microspheres[J]. Jie Pou Xue Yan Jiu, 2008,30(4):241-244
	黄婉丹, 龙大宏, 谷海刚 . BSA-PLGA缓释微球的制备及优化条件的探索[J]. 解剖学研究, 2008,30(4):241-244
[9]	Wang PP, Frazier J, Brem H . Local drug delivery to the brain[J]. Adv Drug Deliv Rev, 2002,54(7):987-1013 doi: 10.1016/S0169-409X(02)00054-6 pmid: 12384318
[10]	Li Q, Fan ZZ, Li XQ, Liu XH, Sun J, Gu W, Heng P, Gao H . Effects of interventional therapy with norcantharidin microsphere on hepatoma in rats and its mechanism[J]. J Chin Integr Med, 2006,4(4):378-383
	李琦, 范忠泽, 李先茜, 刘晓华, 孙珏, 顾伟 , Heng P, 高虹.去甲斑蝥素微球介入治疗大鼠肝癌疗效及其机制研究[J]. 中西医结合学报, 2006,4(4):378-383
[11]	Chen ZK, Lin LW, Yang FD, Weng XH, Cai HJ, Xue ES, Wang Y . Effects of interstitial chemotherapy using thermosensitive gel-coated ricin on hepatoma H22-bearing mice[J]. J Chin Integr Med, 2009,7(5):447-452 doi: 10.3736/jcim20090509
	陈志奎, 林礼务, 杨发端, 翁秀华, 蔡华晶, 薛恩生, 王艳 . 蓖麻毒素温敏型凝胶间质化疗对H22肝癌荷瘤小鼠的影响[J]. 中西医结合学报, 2009,7(5):447-452 doi: 10.3736/jcim20090509
[12]	Shive MS, Anderson JM . Biodegradation and biocompatibility of PLA and PLGA microspheres[J]. Adv Drug Deliv Rev, 1997,28(1):5-24 doi: 10.1016/S0169-409X(97)00048-3
[13]	Allen TM, Cullis PR . Drug delivery systems: entering the mainstream[J]. Science, 2004,303(5665):1818-1822 doi: 10.1126/science.1095833
[14]	Lesniak MS, Upadhyay U, Goodwin R, Tyler B, Brem H . Local delivery of doxorubicin for the treatment of malignant brain tumors in rats. Anticancer Res[J]., 2005,25(6B):3825-3831 doi: 10.3171/jns.2002.97.5.1173 pmid: 16312042

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Concentration (μg/mL)	n	Inhibition rate
Concentration (μg/mL)	n	12 h	24 h	48 h
Control	3	0	0	0
0.5	3	16.71±2.27*	24.36±2.54^△	29.42±1.88^△
1	3	21.85±1.67*	37.82±1.15^△	44.36±2.35^△
2	3	33.82±2.15*	52.37±1.75^△	60.14±1.36^△
3	3	47.39±2.03*	64.43±2.27^△	76.32±2.33^△
4	3	69.46±1.57*	82.34±0.89^△	90.58±1.18^△

Group	n	Tumor volume
Group	n	Before treatment	8 days after treatment	14 days after treatment	20 days after treatment	26 days after treatment
Normal saline	8	256.3±24.7	636.8±94.1	1 616.6±203.6	3 237.9±382.8	5 768.9±442.8
Blank microspheres	8	249.5±23.3	617.2±88.5	1 572.8±217.2	3 229.5±316.3	5 447.5±426.3
CTX	8	261.3±30.2	459.3±78.4*	842.3±205.2*	1 937.7±257.1*	3 637.5±293.2*
CTX-PLGA	8	251.5±28.4	379.2±58.1**^△	603.2±139.3**^△	915.6±251.2**^△	1 219.6±263.7**^△

Group	n	Body weight ($\bar{x}$±s, g)	Tumor weight ($\bar{x}$±s, g)	Inhibition rate (%)
Normal saline	8	27.90±1.43	2.75±0.52	–
Blank microspheres	8	28.05±0.79	2.51±0.39	8.73
CTX	8	28.29±1.29	2.02±0.46*	26.55
CTX-PLGA	8	28.74±0.70	1.31±0.31**^△	52.36

Effects of intratumoral injection of microspheres containing cobra venom cytotoxin on transplanted human hepatoma in nude mice

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