蜂毒이 Glioma Cell에 미치는 效果
Effects of Bee Venom on Glioma Cells
0
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
Objective : Bee venom (BV) has been used for the treatment of inflammatory diseases such as rheumatoid arthritis and relief of pain in Oriental medicine. The two main components of BV are melittin and phospholipase A2 (PLA2). Of these, melittin, the major active ingredient of BV, has been reported to induce apoptosis and to possess anti‐tumor effects. Several studies have established that the agents inducing apoptosis in target organs suppress tumorigenesis. As the other component, PLA2 has been reported to induce neurite outgrowth in PC12 cells. However, there was no report about proliferative effect of BV in neuronal cells. In order to examine the effect of BV on glioma cell, human glioma cell line, U87 was used. Methods : Analysis of proliferation was confirmed by MTT assay. BV increased cell number through dose‐ and duration‐dependent manner and these effects are apparent at a concentration of 10 ug/ml. To observe which signaling molecules will be activated by BV, phosphorylation of Akt, MAPK, PYK2 or CREB were examined by Western blot analysis. To study the long term effect of BV in U87 cells, the image of cells treated with BV for 4 days were obtained. Results : The phosphorylation levels of PYK2 and Akt were increased at 5 min after addition of 10 ug/ml of BV and sustained to 2 hours. On the other hand, phosphorylation of MAPK and CREB were increased at 5 min, maximum at 10 min, and returned to 30 min. These imply that BV may activate two different signaling pathways, PYK2/Akt and MAPK/CREB. BV treated cells showed increased neurite number and length. Conclusion : These results propose that BV may induce differentiation as well as proliferation of U87 cells through the activation of PYK2/Akt and MAPK/CREB.
Abstract
Objective : Bee venom (BV) has been used for the treatment of inflammatory diseases such as rheumatoid arthritis and relief of pain in Oriental medicine. The two main components of BV are melittin and phospholipase A2 (PLA2). Of these, melittin, the major active ingredient of BV, has been reported to induce apoptosis and to possess anti‐tumor effects. Several studies have established that the agents inducing apoptosis in target organs suppress tumorigenesis. As the other component, PLA2 has been reported to induce neurite outgrowth in PC12 cells. However, there was no report about proliferative effect of BV in neuronal cells. In order to examine the effect of BV on glioma cell, human glioma cell line, U87 was used. Methods : Analysis of proliferation was confirmed by MTT assay. BV increased cell number through dose‐ and duration‐dependent manner and these effects are apparent at a concentration of 10 ug/ml. To observe which signaling molecules will be activated by BV, phosphorylation of Akt, MAPK, PYK2 or CREB were examined by Western blot analysis. To study the long term effect of BV in U87 cells, the image of cells treated with BV for 4 days were obtained. Results : The phosphorylation levels of PYK2 and Akt were increased at 5 min after addition of 10 ug/ml of BV and sustained to 2 hours. On the other hand, phosphorylation of MAPK and CREB were increased at 5 min, maximum at 10 min, and returned to 30 min. These imply that BV may activate two different signaling pathways, PYK2/Akt and MAPK/CREB. BV treated cells showed increased neurite number and length. Conclusion : These results propose that BV may induce differentiation as well as proliferation of U87 cells through the activation of PYK2/Akt and MAPK/CREB.
참고문헌 (37)
- 정인태, "봉독약침이 collagen 유발 관절염에 미치는 진통효과 및 그 기전 연구", 대한침구학회지, 22(1), pp.99~108 (2005), 대한침구학회
- 김용석, "봉독약침요법의 항염증 작용에 관한 실험적 연구", 대한침구학회지, 15(1), pp.317~331 (1988)
- 김문호, "봉독요법과 봉침요법", pp.20~ (1992), 한국교육기획
- Kang SS, "The effect of whole bee venom on arthritis", Am J Chin Med, 30(1), pp.73~80 (2002)
- 이홍석, "최근 10년간 국내의 봉독 관련 연구에 대한 고찰", 대한침구학회지, 20(3), pp.14~ (2003), 대한침구학회
- 이광우, "임상신경학", pp.598~599 (2002), 고려의학
- 대한신경외과학회, "신경외과학", pp.171~176 (2001), 대한신경외과학회
- 尹炫珉, "胡桃藥鍼이 人間의 신경교종 細胞에 유발된 低酸素症에 대한 防禦效果", 대한침구학회지, 20(2), pp.14~ (2003), 대한침구학회
- 박상현, "울금의 폐암, 자궁암, 신경교종 및 전립선암에 대한 항암효과 연구", 박사, pp.2006
- Sladowski D, "An improved MTT assay", J. Immunol. Methods, 157, pp.203~207 (1993)
- Marshall NJ, "A critical assessment of the use of microulture tetazolium assays to measure cell growth and function", Growth regul, 5, pp.69~84 (1995)
- Habermann E, "Chemistry, pharmacology and toxicology of bee, wasp and hornet venoms. In Venomous Animals and their Venoms", pp.3~61 (1971), Academic Press
- Assem ES, "Histamine release by MCDP(401), a peptide from the venom of the honey bee", Bri J Pharmacol, 48(2), pp.337~338 (1973)
- 권기록, "봉침에 대한 고찰", 대한침구학회지, 11(1), pp.160~ (1994)
- 인창식, "봉독요법에 대한 한의학 최초의 문헌: 마왕퇴의서의 봉독요법 2례", 대한침구학회지, 15(1), pp.143~147 (1998)
- 郭國華, "臨床中藥辭典", pp.543~544 (1994), 湖南科學技術出版社
- 李文瑞, "中藥別名辭典", pp.1079~ (1994), 中國科學技術出版社
- 성은찬, "난치병의 봉침요법", pp.48~ (1958), 전국농업기술자협회 출판부
- Son DJ, "Melittin inhibits vascular smooth muscle cell proliferation through induction of apoptosis via suppression of nuclear factor-kappaB and Akt activation and enhancement of apoptotic protein expression", J Pharmacol Exp Ther, 317(2), pp.627~634 (2006)
- Hu H, "Effect of polypeptides in bee venom on growth inhibition and apoptosis induction of the human hepatoma cell line SMMC-7721 in-vitro and Balb", c nude mice in-vivo, 2006, pp.58(1), 83~89
- Putz T, "Bee venom secretory phospholipase A2 and phosphatidylinositol - homologues cooperatively disrupt membrane integrity, abrogate signal transduction and inhibit proliferation of renal cancer cells", Cancer Immunol Immunother (2006)
- Jang MH, "Bee venom induces apoptosis and inhibits expression of cyclooxygenase-2 mRNA in human lung cancer cell line NCI-H1299", J Pharmacol Sci, 91(2), pp.95~104 (2003)
- Sato H, "Peptide-membrane interactions and mechanisms of membrane destruction by amphipathic alpha-helical antimicrobial peptides", Biochim Biophys Acta, 1758(9), pp.1245~1256 (2006)
- Nakashima S, "Secretory phospholipases A2 induce neurite outgrowth in PC12 cells", Biochem J, 376(3), pp.655~666 (2003)
- 김기령, "SOOMOONSA`S ENGLISH-KOREAN MEDICAL DICTIONARY", pp.1376~13777 (1985), 수문사
- Shaposhnikova VV, "The effect of melittin on proliferation and death of thymocytes", FEBS Lett, 410(2), pp.285~288 (1997)
- Ghosh A, "Calcium signaling in neurons. molecular mechanisms and cellular consequences", Science, 268, pp.239~247 (1995)
- Chrivia JC, "Phosphorylated CREB binds specifically to the nuclear protein CBP", Nature, 365, pp.855~859 (1993)
- Deisseroth K, "Translocation of calmodulin to the nucleus supports CREB phosphorylation in hippocampal neurons", Nature, 392(6672), pp.198~202 (1998)
- Rivera VM, "A growth factor-induced kinase phosphorylates the serum response factor at a site that regulates its DNA-binding activity", Mol Cell Biol, 13, pp.6260~6273 (1993)
- Lev S, "Protein tyrosine kinase PYK2 involved in Ca(2+)-induced regulation of ion channel and MAP kinase functions", Nature, 376(6543), pp.737~745 (1995)
- Basile JR, "Semaphorin 4D", plexin-B1 induces endothelial cell migration through the activation of PYK2, Src, and the phosphatidylinositol 3-kinase-Akt pathway, 2005, pp.25(16), 6889~6898
- Gonzalez L, "Characterization of melittin effects in synaptosomes", Neurochem Res, 22(2), pp.189~199 (1997)
- Lee SY, "Melittin exerts multiple effects on the release of free fatty acids from L1210 cells: lack of selective activation of phospholipase A2 by melittin", Arch Biochem Biophys, 389(1), pp.56~57 (2001)
- pyrene diol epoxide (BPDE) in mammary epithelial cells, "Carcinogenesis", 2006, 2331~2340
- Spoerri PE, "Changes induced by apamin from bee venom on differentiated mouse neuroblastomacells", culture Acta Anat (Basel), 117(4), pp.346~354 (1983)
- Kwok RP, "Nuclear protein CBP is a coactivator for the transcription factor CREB", Nature, 370, pp.223~226 (1994)