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探討鳳梨酵素藉由調控細胞凋亡蛋白機轉抑制大腸癌細胞研究

The Inhibitory Effect of Colon Cancer Cells by Bromelain via Modulate the Mechanism of Apoptosis Proteins

摘要


目的:鳳梨酵素經研究證實有抗氧化、抗血栓、傷口癒合及免疫調節為主;在動物實驗中亦證實具有包括清除活性氧自由基、清除活化態的致癌劑或致突變劑、調節細胞週期蛋白、改變基因表達、提高免疫力等,然而對抗大腸癌效果則未曾有系統的研究,臨床上大腸癌的發生率近年來有顯著上升的趨勢,因此如何預防與治療大腸癌已成為重要的課題。方法:本計劃以HT29(Human colon adenocarcinoma grade II cell line;人類結腸直腸腺癌細胞株)進行體外培養,探討大腸癌化中鳳梨酵素對於細胞凋亡(apoptosis)中Bcl-xL、Bax、PARP與Cyt-C訊息路徑的影響,並且將以流式細胞儀分析癌細胞之細胞週期變化、細胞凋亡以及使用西方墨點法細胞凋亡知相關訊息路徑變化情形與下游蛋白質的表現。以進一步探討鳳梨酵素抑制大腸癌細胞之機轉。結果:研究結果顯示,細胞再加入不同濃度之鳳梨酵素後,經MTT分析後發現鳳梨酵素使大腸癌細胞之生長受阻,且經flow cytometry分析後發現鳳梨酵素亦明顯誘發細胞凋亡反應,顯示鳳梨酵素確實具有促使大腸癌細胞凋亡之能力,接下來本研究進行細胞凋亡蛋白的分析,在運用西方墨點法(western blots)進一步檢測相關蛋白質後,研究結果發現細胞凋亡抑制蛋白Bcl-xL在經過鳳梨酵素處理過後有明顯被抑制的現象,而細胞凋亡蛋白Bax、PARP與Cyt-C則皆出現明顯上升趨勢。研究結果顯示鳳梨酵素確實具有促使大腸癌細胞凋亡之能力,且透過誘發細胞凋亡蛋白Bax、PARP與Cyt-C與抑制細胞凋亡抑制蛋白Bcl-xL含量來達成誘發細胞凋亡反應。結論:因此本研究成功證實鳳梨酵素具有抗大腸癌細胞之能力與其可能之抗癌作用機轉,本研究不止促進抗癌細胞相關機制的瞭解,亦可開發另一種抗癌之輔助療法。

關鍵字

鳳梨酵素 大腸癌 細胞凋亡

並列摘要


Objective: Studies indicate that bromelain exerts antioxidant, anticancer and immunomodulatory effects, and also proved be capable of free radicals cleavage, anti- carcinogenicity, apoptosis proteins modulation in animal models. The anti- cancer effects by bromelain against colon cancer were never been systemic evaluated. The purpose of this study is to verify the inhibitory effect on colon carcinogenicity by bromelain. Methods: In this study, we used bromelain to inhibit colon cancer cell proliferation and tumor growth, and investigate the underlying molecular mechanisms. HT-29 cell line (human colon adenocarcinoma cell line) was used in this study. Cell cycles were analyzed by flow cytometry. Signal transductions were analyzed by western blots. Results: The results indicated that the viability of HT-29 cells were reduced by MTT assay and the apoptosis of HT-29 cells were significantly in duced by flow cytometry. The results indicated the amounts of bax、parpand Cyt-C and parp were dose-related induced, but the amounts of BCL-xL, the inhibitor of apoptosis, were dose-related decreased. Conclusions: The results of present study can provide the experimental evidence that inhibitory effect on anti- carcinogenicity of colorectal adeno- carcinoma by bromelain.

並列關鍵字

bromelain signal transduction apoptosis

延伸閱讀