Anti-tumor activities and apoptosis-regulated mechanisms of bufalin on the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice
Anti-tumor activities and apoptosis-regulated mechanisms of bufalin on the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice
复制标题
蟾蜍灵对人肝癌裸鼠原位移植瘤模型的抗肿瘤活性及凋亡调节机制
DOI：
10.3748/wjg.v13.i24.3374
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发表时间：
2007-06-28
影响因子：
4.3
通讯作者：
Ling, Chang-Quan
中科院分区：
文献类型：
--
作者：
Han, Ke-Qi;Huang, Guang;Ling, Chang-Quan
目的：目的：探讨蟾蜍灵对人肝癌裸鼠原位移植瘤的抑制作用及其调控机制。方法：将人肝癌BEL-7402细胞接种于裸鼠皮下形成肿瘤，然后植入裸鼠肝脏，建立人肝癌裸鼠原位移植瘤模型。将75只动物随机分为5组（n = 15）。蟾蜍灵1.5mg/kg（BF 1）、1 mg/kg（BF 2）、0.5mg/kg（BF 3）腹腔注射，连续15-24 d。NS组注射等量生理盐水，ADM组腹腔注射阿霉素8.0mg/kg，连续15 d。在第25天每组处死10只小鼠，计算各组小鼠的存活时间。通过病理学和电镜观察心肌、脑、肝、肾及肿瘤组织的形态学改变，TUNEL染色法检测肿瘤组织细胞凋亡率，免疫组化染色和RT-PCR法检测肿瘤组织中凋亡调控基因bcl-2和bax的表达。蟾毒灵各剂量组肿瘤体积均明显缩小（35.21 +/- 12.51 vs 170.39 +/- 25.29; 49.83 +/- 11.46 vs 170.39 +/- 25.29; 83.99 ± 24.63 vs 170.39 ± 25.29，P < 0.01），BF 1 -2组存活时间延长（分别为31.8 +/- 4.2 vs 23.4 +/- 2.1和29.4 +/- 3.4 vs 23.4 +/- 2.1，P < 0.05），BF 1 -2组以重度和中度坏死为主。心肌、脑、肝、肾组织未见形态学改变。BF 1 -2组可见凋亡特征。免疫组化检测各组bcl-2、bax蛋白表达阳性率分别为10.0%、10.0%、20.0%、10.0%、20.0%; 90.0%、80.0%、80.0%、40.0%、30.0%。结论：蟾蜍灵对人肝癌裸鼠原位移植瘤有明显的抗肿瘤作用，无明显毒性，并能诱导移植瘤细胞凋亡。这种凋亡可能主要是通过上调凋亡调控基因bax的表达而介导的，这可能参与了蟾蜍灵的抗肿瘤作用机制。(c)2007年，WIG出版社。All rights reserved.
AIM: To investigate anti-tumor activities and apoptosis-regulated mechanisms of bufalin in the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice.METHODS: BEL-7402 cells of human hepatocellular carcinoma were inoculated to form subcutaneous tumors, and were implanted into the liver to establish orthotopic transplantation tumor models of human hepatocellular carcinoma in nude mice. Seventy-five animals were randomized divided into five groups (n = 15). Bufalin was injected intraperitoneally into three groups at doses of 1.5 mg/kg (BF1), 1 mg/kg (BF2) and 0.5 mg/kg (BF3) for d 15-24, respectively. The NS group was injected an equal volume of saline as above and adriamycin was injected intraperitoneally into the ADM group at a dose of 8.0 mg/kg for d 15. Ten mice in each group were killed at d 25 and the survival time in each group was calculated. We also observed the morphologic alterations in the myocardium, brain, liver, kidney and tumor tissues by pathology and electron microscopy, measured the apoptotic rate by TUNEL staining method, and detected the expression of apoptosis-regulated, genes bcl-2 and bax by immunohistochemical staining and RT-PCR in tumor tissues.RESULTS: The tumor volumes in each group of bufalin were reduced significantly (35.21 +/- 12.51 vs 170.39 +/- 25.29; 49.83 +/- 11.46 vs 170.39 +/- 25.29; 83.99 +/- 24.63 vs 170.39 +/- 25.29, P < 0.01, respectively), and the survival times were prolonged in group BF1-2 (31.8 +/- 4.2 vs 23.4 +/- 2.1 and 29.4 +/- 3.4 vs 23.4 +/- 2.1, P < 0.05, respectively), and necrosis was mainly in severe or moderate degree in group BF1-2. No morphological changes were detected in the myocardium, brain, liver and kidney tissues. Apoptotic characteristics could be seen in group BF1-2. The positive rates of bcl-2 and bax protein expression of each group by immunohistochemical staining were 10.0%, 10.0%, 20.0%, 10.0% and 20.0%; 90.0%, 80.0%, 80.0%, 40.0% and 30.0%, respectively. Loss of expression of bcl-2 mRNA in each group was to be found and the density of bax mRNA was increased progressively with increase of dose of bufalin by RT-PCR.CONCLUSION: Bufalin has significant anti-tumor activities in the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice with no marked toxicity and was able to induce apoptosis of transplanted tumor cells. This apoptosis may be mediated mainly via up-regulating the expression of apoptosis-regulated gene bax, which may be involved in its anti-tumor mechanism of bufalin. (c) 2007 The WIG Press. All rights reserved.