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Journal of Chinese Integrative Medicine ›› 2007, Vol. 5 ›› Issue (2): 155-159.doi: 10.3736/jcim20070211

• Original Experimental Research • Previous Articles     Next Articles

Inhibition action of bufalin on human transplanted hepatocellular tumor and its effects on expressions of Bcl-2 and Bax proteins in nude mice

Wei Gu, Ke-qi Han, Yong-hua Su, Xue-qiang Huang, Chang-quan Ling   

  1. Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
  • Online:2007-03-31 Published:2007-03-15

Objective: To investigate the anti-tumor effect of bufalin and its regulation on Bcl-2 and Bax proteins in orthotopically transplanted tumor of human hepatocellular carcinoma in nude mice. 

Methods: Orthotopically transplanted tumor of human hepatocellular carcinoma was established in nude mice. The mice were randomly divided into five groups: high-dose bufalin-treated group (1.5 mg/kg), medium-dose bufalin-treated group (1 mg/kg), low-dose bufalin-treated group (0.5 mg/kg), adriamycin-treated group (8.0 mg/kg), and normal saline-treated group. After 25 days, mice were sacrificed. The tumor volume was measured, and the pathological changes of tumor tissues were detected by HE staining to observe the tumor necrosis degree. Cell morphological changes were also observed by an electron microscopy. Label index of tumor cell apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and the expressions of Bcl-2 and Bax proteins were determined by immunohistochemical method. 

Results: The tumor volume in the bufalin-treated groups was shrunk significantly compared with that in the normal saline-treated group (P<0.01). The survival time of the bufalin-treated groups was prolonged compared with that of the adriamycin-treated group and the normal saline-treated group (P<0.05). Apoptotic characteristics could be seen in tumor tissues of the bufalin-treated groups. The label index of tumor cell apoptosis in the bufalin-treated groups (5.87±2.13, 8.86±2.96 and 10.60±3.42 in low-, medium- and high-dose groups respectively) was higher than that in the adriamycin-treated group (3.28 ±0.98) (P<0.05, P<0.01). The expression of Bax was up-regulated, while no changes were detected as to Bcl-2 protein in tumors of the bufalin-treated groups.

Conclusion: Bufalin has significant anti-tumor effect on the orthotopically transplanted tumor of human hepatocellular carcinoma in nude mice. Its effect might be related to up-regulation of Bax protein and inducement of the tumor cell apoptosis.

Key words: bufalin, liver neoplasms, apoptosis, Bcl-2, Bax, nude mice

CLC Number: 

  • R735.7

Table 1

Changes of mouse body weight and tumor volume of human hepatocellular orthotopically transplanted tumor induced by bufalin ($\bar{x}±S$)"

GroupnBody weight (g)Tumor volume (mm3)Tumor inhibition rate
Pre-treatmentPost-treatmentPre-treatmentPost-treatment
High-dose bufalin-treated1020.94±1.0021.39±1.62*43.50±5.0935.21±12.51△△79.3%*
Medium-dose bufalin-treated1020.03±1.1621.48±1.10*42.92±4.1049.83±11.46△△ 70.70%
Low-dose bufalin-treated1021.06±0.9421.57±1.14*43.00±2.9783.99±24.63△△ 50.70%
ADM-treated1021.00±1.0018.90±0.7742.93±4.2355.17±16.13△△ 67.60%
NS-treated1020.95±1.0720.40±1.2343.37±4.82170.39±25.291

Figure 1

Mean survival time of five groupsBu1: High-dose bufalin-treated group; Bu2: Medium-dose bufalin-treated group; Bu3: Low-dose bufalin-treated group; *P<0.05, vs NS-treated group; n=5"

Figure 2

Pathological characteristics of tumor necrosi (HE staining, ×250)A: NS-treated group; B: High-dose bufalin-treated group."

Table 2

Label index of tumor cell apoptosis and expressions of Bcl-2 and Bax in tumor tissues ($\bar{x}±S$)"

GroupnLabel index (%)
ApoptosisBcl-2Bax
High-dose bufalin-treated1010.60±3.42**8.76±5.1890.70±16.82**
Medium-dose bufalin-treated108.86±2.96**8.52±3.5780.21±21.23**
Low-dose bufalin-treated105.87±2.13*11.34±6.2479.54±10.75**
ADM-treated104.26±2.1210.59±6.9247.56±15.31*
NS-treated103.28±0.989.05±4.9731.25±12.50

Figure 3

Expression of Bax in orthotopically transplanted tumor of humanhepatocellular carcinoma in nude mice (SP method, ×200) A: Bax expressed weakly in NS-treated group. B: High expression in high-dose bufalin-treated group."

[1] Yamada K, Hino K, Tomoyasu S.et al.Enhancement by bufalin of retinoic acid induced differentiation of acute promyelocytic leukemia cells in primary culture[J].Leuk Res,1998,22(7):589-595
Yamado K, Hino K, Tomoyasu S.et al.Enhancement by bufalin of retinoic acid induced differentiation of acute promyelocytic leukemia cells in primary culture[J].Leuk Res,1998,22(7):589-595
[2] Yeh JY, Huang WJ, Kan SF, et al.Effects of bufalin and cinobufagin on the proliferation of androgen dependent and independent prostate cancer cells[J].Prostate,2003,54(2):112-124
[3] 陈小义, 徐瑞成, 陈莉, 等.蟾蜍灵诱导人胃癌细胞凋亡的实验研究[J].基础医学与临床,2000,20(5):50-53
Chen XY, Xu RC, Chen L, et al.Apoptosis of gastric cancer cells induced by bufalin[J].Ji Chu Yi Xue Yu Lin Chuang,2000,20(5):50-53
[4] 顾伟, 韩克起, 苏永华, 等.蟾毒灵对人肝癌BEL-7402细胞增殖的影响[J].中西医结合肝病杂志,2006,16(1):16-17, 20
Gu W, Han KQ, Su YH, et al.Effect of bufalin on proliferation of BEL-7402 human hepatocarcinoma cell line[J].Zhong Xi Yi Jie He Gan Bing Za Zhi,2006,16(1):16-17, 20
[5] 顾伟, 沈婕, 翟笑枫.大鼠移植性肝癌模型生物学行为与分期的初步探讨[J].中西医结合学报,2005,3(2):136-138
Gu W, Shen J, Zhai XF.Preliminary study on biological behavior and staging of transplanted hepatoma in rats[J].Zhong Xi Yi Jie He Xue Bao,2005,3(2):136-138
[6] 韩克起, 顾伟, 张亚妮, 等.裸鼠人肝癌原位移植模型的建立[J].第四军医大学学报,2004,25(17):
Han KQ, Gu W, Zhang YN, et al.Establishment of orthotopic transplantation tumor models of human hepatocellular carcinoma in nude mice[J].Di Si Jun Yi Da Xue Xue Bao,2004,25(17):inside front cover
[7] 李琦, 范忠泽, 李先茜, 等.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
Li Q, Fan ZZ, Li XQ, et al.Effects of interventional therapy with norcantharidin microsphere on hepatoma in rats and its mechanism[J].Zhong Xi Yi Jie He Xue Bao,2006,4(4):378-383
[8] 陈喆, 翟笑枫, 苏永华, 等.华蟾素注射液治疗中晚期原发性肝癌临床疗效观察[J].中西医结合学报,2003,1(3):184-186
Chen Z, Zhai XF, Su YH, et al.Clinical observation of cinobufacini injection used to treat moderate and advanced primary liver cancer[J].Zhong Xi Yi Jie He Xue Bao,2003,1(3):184-186
[9] Kamano Y, Nogawa T, Yamashita A, et al.Isolation and structure of a 20, 21-epoxybufenolide series from ''Ch'an Su''[J].J Nat Prod,2002,65(7):1001-1005
doi: 10.1194/jlr.M900181-JLR200 pmid: 12141860
[10] 苏永华, 牛欣.蟾酥制剂的药效作用研究评述[J].北京中医药大学学报,2001,24(2):51-54
Su YH, Niu X.Pharmacodynamic research of ''Ch'an Su'' praeparatum: a review[J].Beijing Zhong Yi Yao Da Xue Xue Bao,2001,24(2):51-54
[11] 苏永华, 尹西才, 谢觉民, 等.三种蟾毒单体对SMMC-7721和BEL-7402人肝癌细胞生长的抑制作用[J].第二军医大学学报,2003,24(4):393-395
Su YH, Yin XC, Xie JM, et al.Inhibition effects of three kinds of bufotoxins on human SMMC-7721 and BEL-7402 hepatoma cell lines[J].Di Er Jun Yi Da Xue Xue Bao,2003,24(4):393-395
[12] 江苏新医学院.中药大辞典[M].上海: 上海科学技术出版社,1977,():84-85
Jiangsu New Medical College.Dictionary of Chinese Traditional Drugs[M].Shanghai: Shanghai Scientific and Technical Publishers,1977,():84-85
[13] Cory S, Adams JM.Killing cancer cells by flipping the Bcl-2/Bax switch[J].Cancer Cell,2005,8(1):5-6
doi: 10.1016/j.ccr.2005.06.012 pmid: 16023593
[14] Terada T, Nakanuma Y.Expression of apoptosis, proliferating cell nuclear antigen, and apoptosis-related antigen(bcl-2, c-myc, Fas, Lewis(y) and p53) in human cholangiocarcinomas and hepatocellular carcinomas[J].Pathol Int,1996,46(10):764-770
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