Energi-702對人類惡性骨肉瘤細胞的抗癌機轉研究

Investigating anti-tumor effects of Energi-702 on human osteosarcoma cells

陳金釧(Kim Chuan Chan)

指導教授 ： 高紹軒

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2013年)

摘要

AMP-活化蛋白質激酶 (AMPK) 調控細胞內多項生理訊息傳遞路徑並參與多項生理調控。過去文獻指出，活化AMPK具有抑制正常細胞及癌細胞生長的能力，也具有治療癌症的潛力。本篇論文研究主要在探討Energi-702（AMPK 活化劑）對於人類骨肉瘤細胞（U2OS、143B）的抗癌機轉。首先利用MTT觀察細胞在加藥處理後的存活率以及利用電子顯微鏡觀察細胞的形態變化。接下來把惡性骨肉瘤細胞處理24小時後，收集細胞蛋白質萃取液進行西方墨點法觀察，發現與細胞週期相關蛋白質cyclin D1、cyclin E、Cdk 2及Cdk 4在Energi-702的作用下都被抑制。此外我們也發現處理Energi-702後，腫瘤抑制因子蛋白質p27及 p53有明顯的增加。最後我們也觀察了在Energi-702的作用下細胞凋亡相關蛋白質的表現量，發現caspase-3,、caspase-9、 及 PARP 都有被活化的現象，而抗凋亡蛋白Bcl-2亦有顯著的下降。綜合實驗結果，證明Energi-702除了影響U2OS、143B細胞生長使細胞週期的停滯也促使細胞凋亡。

關鍵字

人類惡性骨肉瘤細胞

並列摘要

AMP-activated protein kinase (AMPK) is a key player in maintaining energy homeostasis in response to metabolic stress. Published studies indicate that AMPK activation strongly suppresses cell proliferation in
non-malignant cells as well as in tumour cells. This study examined the anti-tumor mechanism of Energi-702 (AMPK activator) on osteosarcoma cell lines (U2OS & 143B). Cell viability was detected by the MTT assay and cell morphology was observed after drug administration. After treated with Energi-702 for 24 hour, osteosarcoma cell lysates were collected. Western blot results demonstrate that cell cycle-associate proteins cyclin D1, cyclin E, cdk 2 and cdk 4 were inhibited after Energi-702 treatment. We also found that tumor suppressor gene p27 and p53 have significant elevation after Energi-702 treatment. On the other hand, the western blot result showed the activation of caspase-3, caspase-9, and PARP by Energi-702. In parallel, Energi-702 significantly decreased the level of anti-apoptotic protein, Bcl-2. Taken together, these findings indicate that Energi-702 diminished cell viability through cell cycle arrest and apoptosis.

並列關鍵字

osteosarcoma cells

參考文獻

1. Broadhead, M.L., et al., The molecular pathogenesis of osteosarcoma: a review. Sarcoma, 2011. 2011: p. 959248.

3. Cotterill, S.J., et al., Stature of young people with malignant bone tumors. Pediatr Blood Cancer, 2004. 42(1): p. 59-63.

4. Gelberg, K.H., et al., Growth and development and other risk factors for osteosarcoma in children and young adults. Int J Epidemiol, 1997. 26(2): p. 272-8.

5. Lau, W.M., et al., Enpp1: a potential facilitator of breast cancer bone metastasis. PLoS One, 2013. 8(7): p. e66752.

6. Goyal, S., et al., Symptom interval in young people with bone cancer. Eur J Cancer, 2004. 40(15): p. 2280-6.