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

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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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