葫蘆素B抑制大腸直腸癌DLD-1細胞 之角色探討

Inhibitory effects of Cucurbitacin B on human DLD-1 colon adenocarcinoma cells

林雨潔(Yu-Jie Lin)

指導教授 ： 曾博修

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2011年)

https://doi.org/10.6834/CSMU.2011.00042

摘要

Cucurbitacin B是三萜類化合物(triterpenoid)的一種，文獻指出Cucurbitacin B具有抗發炎、抗癌的功能，能產生細胞毒殺性抑制人類癌症細胞的增生，最後誘導癌症細胞凋亡的產生。因此，我們用Cucurbitacin B處理大腸直腸癌細胞DLD-1，並探討其抑制大腸直腸癌細胞增生之機轉。透過MTT assay在不同時間、不同劑量的Cucurbitacin B處理大腸直腸癌細胞DLD-1，測定癌細胞增殖抑制的情形，則Cucurbitacin B有效的抑制大腸直腸癌細胞DLD-1的增生；利用倒立顯微鏡觀察Cucurbitacin B處理後大腸直腸癌細胞型態的變化，造成癌細胞型態轉為皺縮；透過西方墨點法觀察Cucurbitacin B處理之後對凋亡蛋白Apaf-1、cytochrome C、Caspase-3及促凋亡蛋白Bad、Bax、t-bid的影響，MAPKs路徑相關蛋白，包括pErk、Erk、pP38、P38、pJNK、JNK (c-Jun N-terminal kinase)表達的影響，及對TNF-α、Fas、Fas-Ligand、STAT3、PI3K的影響。結果發現Cucurbitacin B影響Apaf-1、cytochrome C表現增加，Caspase-3也被活化；進一步的研究發現，Cucurbitacin B處理之後TNF-α表現量增加及JNK磷酸化表現量也增加。因此，Cucurbitacin B對於大腸直腸癌細胞DLD-1具有明顯的細胞毒殺作用，能誘導DLD-1細胞凋亡，可能也有透過TNF-α表現、JNK的激活機制參與細胞凋亡的其中。

關鍵字

葫蘆素B

並列摘要

Cucurbitacin B , a natural triterpenoid is well Known for its anti-inflammatory and anti-cancer activity.It can antiproliferative 、cytotoxic anticancer activity and induce apopotosis in several human caner cells.The mechanisms of anti-proliferation induced by Cucurbitacin B in human colon cancer cells are not unknown , we therefore treated colon cancer cell DLD-1 with Cucurbitacin B. To determineinhibition of cell proliferation with MTT assay, at different times and different doses of Cucurbitacin B treatment of colon cancer cells DLD-1 , then Cucurbitacin B colon cancer cells effectively inhibit the proliferation of DLD-1. After treatment Cucurbitacin B ,cell morphology changes Useing microscope and the cells became round and refractile.By Western blot , Cucurbitacin B after processing of the apoptotic protein Apaf-1, cytochrome C, Caspase-3 effects, Bad,Bax,t-Bid and MAPKs path-related proteins, including pErk, Erk, pP38, P38, pJNK, JNK(c-Jun N-terminal kinase) expression, and of TNF-α, Fas, Fas-Ligand, STAT3, PI3K effects. Cucurbitacin B affect Apaf-1, cytochrome C performance increase, Caspase-3 is also activated; further , after treatment Cucurbitacin B increased TNF-α expression and JNK phosphorylation also increased. Therefore, Cucurbitacin B has significant cytotoxicity for colorectal cancer cells DLD-1 cells ,and induced DLD-1 cells apoptosis.That may also have through TNF-α, JNK activation induced cell apoptosis.

並列關鍵字

Cucurbitacin B

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