| 摘要: | 蜂肽（Bee venom）是一種在藥理學及生物學上具有多種活性的複合物，在傳統的東方醫學中已被運用在減輕疼痛及治療發炎性疾病上如：類風濕性關節炎。最近的研究報告指出蜂肽具有抗腫瘤及誘導癌細胞凋亡作用。本研究中藉由探討細胞增生（proliferation）、細胞週期（cell cycle）、細胞凋亡（apoptosis）及轉移（metastasis）等相關機制，評估蜂肽（Bee venom）對人類子宮頸癌上皮細胞株（Ca Ski）之抗癌作用。由實驗結果發現，蜂肽在Ca Ski細胞上有抑制增生的作用且具有劑量及時間上效應，但對於正常胸主動脈平滑肌A10增生抑制作用不明顯。蜂肽會誘導 Ca Ski 細胞週期停滯在G0/G1期及產生細胞凋亡。經蜂肽處理Ca Ski不僅會造成細胞型態改變，經由DAPI、comet assay、DNA fragmentation分析都發現蜂肽會造成Ca Ski細胞發生DNA damage。利用流式細胞儀分析證實蜂肽作用於Ca Ski細胞株在短時間內會造成ROS及calcium釋放而影響粒腺體膜電位（mitochondrial membrane potential；MMP）誘導細胞凋亡。DNA damage後會活化p53，p53會進一步活化p21及Bax，一方面p21、p27活化後會抑制cyclin E-CDK2 complex；蜂肽也會造成p18、p19活化而抑制cyclin D-CDK4/6 complex因此造成細胞週期停滯在G0/G1期。另一方面在粒腺體凋亡路徑中Bax、Bad活化後會抑制Bcl-2表現而影響粒腺體膜電位使cytochrome c釋放，活化caspase-9進一步活化caspase-3、7而使細胞產生凋亡。由結果也發現蜂肽會增加Fas/FasL表現活化caspase-8，進一步活化caspase-3及造成Bid裂解而使細胞走向凋亡。在MAPK family蜂肽會造成ERK1/2活性下降，JNK1、p38活性上升。另外蜂肽也會降低轉移相關酵素matrix metalloproteinase-2、-9蛋白表現，而抑制細胞轉移。綜合以上的結果發現蜂肽會活化粒腺體路徑和死亡受體接受器路徑使Ca Ski 細胞株產生凋亡。
Bee venom is a complex with pharmacology and biological activity, and it has been used for the treatment of inflammatory diseases such as Rheumatoid arthritis and relief of pain in Oriental medicine. Recently study indicated that bee venom has anticancer effect and induced tumor apoptosis. In current study, we investigated the potential of bee venom on anticancer effect, such as regulation of proliferation, cell cycle progression, apoptosis and metastasis by human cervical epidermoid carcinoma cell line (Ca Ski). The results indicate that bee venom inhibited proliferation of Ca Ski cells in the concentration-dependant and time-dependent management but had no effect been observed in aorta abdominalis smooth muscle-A10. Bee venom also induced cell cycle arrest on G0/G1 phase and apoptosis in Ca Ski cells. We observed morphological changes, DNA fragmentation by comet assay and DAPI method these indicated that bee venom induced Ca Ski cells DNA damage. The results of Flow cytometry have demonstrated that bee venom influence mitochondrial membrane potential by inducing ROS generation and calcium release. p53 could be activated after DNA damage then activate p21 and Bax. Furthermore, p21 and p27 protein will down-regulate cyclin E-CDK 2 complex, p18 and p19 protein will down-regulate cyclin D3-CDK4/6 complex to cause cell cycle arrest on G0/G1. Make mention of the mitochondrial pathway, bee venom changed mitochondrial membrane potential by increasing the Bax and Bad protein expression, inhibiting Bcl-2 protein production to lead cytochrome c release from mitochondria and activate caspase-9, up-regulated caspase-3, -7 to cause apoptosis further. Additionally, bee venom up-regulated the expression of Fas/FasL protein , caspase-8 and truncated Bid. Abut MAPK family, bee venom could down-regulate ERK1/2 and up-regulate JNK1, p38. Bee venom also reduced expression of matrix metalloproteinase-2,-9 and inhibited metastasis. Therefore, the bee venom could induce apoptosis by activating mitochondrial pathway and death-receptor pathway in Ca Ski cell line. |
