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Traditional Medicine Research  2018, Vol. 3 Issue (1): 22-28    DOI: 10.12032/TMR201809061
Special Therapy of Traditional Medicine     
Anti-angiogenesis effect of melittin on Mock/MHCC97-H cells by the regulation of cathepsin S in vivo
Guang-Qiang Ye1, Zhi Zhang2, Chun-Hui Ye1, Keooudone Thammavong1, Jing Xu1,*()
1Department of Hepatobiliary Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China.
2Department of Hepatobiliary Surgery, the First People’s Hospital of Nanning, Nanning, Guangxi, China.
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Highlights

Melittin can inhibit the growth of tumors and angiogenesis by blocking the Cathepsin S-VEGF-A signaling pathway.

Editor’s Summary

The earliest literature recording on bee venom appeared in the book of Jishenlu, which published in Song Dynasty by Xu Xun (916 A.D. - 991 A.D.) and referenced the use of bee venom in the treatment of rheumatism.

Abstract

Objective: To study the anti-angiogenesis effect of melittin on human hepatoma Mock/MHCC97-H cells by regulating the expression of cathepsin S (CatS) in vivo. Methods: Models of in situ transplantation tumor of Mock/MHCC97-H cells and silencing cathepsin shRNA-CatS/ MHCC97-H cells in nude mice were established. The model mice were randomly divided into four groups. In the A1 group, the mice were inoculated with shRNA-CatS/MHCC97-H cells and treated with melittin. In the A2 group, the mice were inoculated with shRNA-CatS/MHCC97-H cells and treated with saline. In the B1 group, the mice were inoculated with Mock/MHCC97-H cells and treated with melittin. In the B2 group, the mice were inoculated with Mock/MHCC97-H cells and treated with saline. The A1 and B1 group were injected with melittin (80 mg/kg) intraperitoneally every day. The A2 and B2 group were injected with 0.2 mL normal saline intraperitoneally every day. After administration for 25 days, the animals were sacrificed. The tumor size and weight in nude mice in each group were recorded. The expression of CD34 protein in the xenograft tumor tissues was detected by immunohistochemistry. The expression of Cat S, VEGF-A, p-VEGFR2, Ras, Raf, p-Raf, MEK1, p-MEK1, ERK1/2 and p-ERK1/2 proteins were detected by western blot. Results: The B1 group had significantly smaller tumor volumes and lower tumor weights than the B2 group (both P < 0.001). There was no significant difference between the A1 group and A2 group in tumor volumes and weights. The number of CD34-positive microvessels in the B2 group was significantly higher than that in the A2 group (P < 0.001). The number of CD34-positive microvessels in the B1 group was significantly lesser than that in the A1 group (P < 0.001). Most strikingly, in the model featuring inoculation of Mock/MHCC97-H cells, CatS, VEGF-A, p-VEGFR2, Ras, Raf, p-Raf, MEK1, p-MEK1, ERK1/2 and p-ERK1/2 expression were inhibited when treated with melittin. However, in the model featuring the inoculation of shRNA-CatS/MHCC97-H cells, no such effects were observed with similar treatments. Conclusion: Melittin can inhibit the growth of tumors and angiogenesis by blocking the CatS-VEGf-A signaling pathway.



Key wordsMelittin      Cathepsin S      Human      Liver cancer      Mock/MHCC97-H cells     
Published: 05 January 2018
Fund:  This work was supported by grants from the National Science Foundation of China (No.81360372)
Corresponding Authors: Xu Jing     E-mail: jxuapr@aliyun.com
Cite this article:

Guang-Qiang Ye, Zhi Zhang, Chun-Hui Ye, Keooudone Thammavong, Jing Xu. Anti-angiogenesis effect of melittin on Mock/MHCC97-H cells by the regulation of cathepsin S in vivo. Traditional Medicine Research, 2018, 3(1): 22-28. doi: 10.12032/TMR201809061

URL:

https://www.tmrjournals.com/tmr/EN/10.12032/TMR201809061     OR     https://www.tmrjournals.com/tmr/EN/Y2018/V3/I1/22

Figure 1 Tumor weight comparison

a: A2 group vs B2 group, P < 0.001; b: B1 group vs B2 group, P < 0.001.

A1 Group: The nude mice were inoculated with shRNA-CatS/MHCC97-H cells and treated with melittin; A2 Group: The nude mice were inoculated with shRNA-CatS/MHCC97-H cells and treated with normal saline; B1 Group: The nude mice were inoculated with Mock/MHCC97-H cells and treated with melittin; B2 Group: The nude mice were inoculated with Mock/MHCC97-H cells and treated with normal saline.

Figure 2 Tumor volume comparison

a: A2 group vs B2 group, P < 0.001; b: B1 group vs B2 group, P < 0.001.

A1 Group: The nude mice were inoculated with shRNA-CatS/MHCC97-H cells and treated with melittin; A2 Group: The nude mice were inoculated with shRNA-CatS/MHCC97-H cells and treated with normal saline; B1 Group: The nude mice were inoculated with Mock/MHCC97-H cells and treated with melittin; B2 Group: The nude mice were inoculated with Mock/MHCC97-H cells and treated with normal saline.

Figure 3 Expression of CD34 in tumor tissues (400×)
Figure 4 Expression of CD34 comparison of tumor tissues

b: A2 group vs B2 group, P < 0.001; a: A1 group vs B1 group, P < 0.001.

A1 Group: The nude mice were inoculated with shRNA-CatS/MHCC97-H cells and treated with melittin; A2 Group: The nude mice were inoculated with shRNA-CatS/MHCC97-H cells and treated with normal saline; B1 Group: The nude mice were inoculated with Mock/MHCC97-H cells and treated with melittin; B2 Group: The nude mice were inoculated with Mock/MHCC97-H cells and treated with normal saline.

Figure 5 Expression of CATS and VEGF-A/VEGFR2 /MEKl/ERKl/2 pathway related proteins
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