探討山葵及荖葉萃取物對肝癌與桑葚對膀胱癌細胞毒性其程式性死亡相關基因之表現

Study of the expression of programmed cell death related genes in the cytotoxicity of Wasabia and Piper betle extracts on liver cancer cells and Mulberry extract on bladder cancer cells.

張瑋烈(Wel-Line Chang)

指導教授 ： 周芬碧

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2012年)

https://doi.org/10.6834/CSMU.2012.00106

摘要

壹.中文摘要
膀胱癌跟肝癌其早期的症狀不易被發現，所以發現時都已是發生轉移或是已到了癌症末期。而程式性死亡(包括凋亡跟自噬凋亡)的調控或是執行發生問題是癌症的成因。而程式性死亡的有許多基因的參與，
這兩種程式性死亡的途徑彼此都會產生相互影響，是非常複雜的過程。在之前的研究已證實了桑葚具有抗氧化及抗發炎的效果，山葵有抑制癌細胞增生的效果，而荖葉具有抗氧化及抑制一些癌症產生的效果。而且在本實驗室已經用MTT assay確認桑葚水萃取物對TSGH-8301 (膀胱癌細胞)及山葵跟荖葉水萃物對Hep3B、HepG2 (肝癌細胞)具有毒殺效果，也藉由流式細胞儀去證實這些水萃物都會對細胞週期產生影響，因此想使用real-time PCR的分析去觀察Apoptosis及Autophagy的相關mRMA的表現量。結果證實桑葚水萃物會使TSGH-8301細胞的Apoptosis相關基因中FADD及Caspas-3有上升的表現，但再配合紫杉醇時不論是對pro-apoptosis或是pro-autophagy相關基因並無顯著的影響，所以無法推定是走Apoptosis或是Autophagy的途徑。而山葵水萃物對於HepG2細胞會使Apoptosis相關基因中的FASLG及Caspas-3有上升表現。但在Hep3B細胞不論是山葵水萃物或是荖葉水萃物對於pro-apoptosis或是pro-autophagy都無顯著性的表現，因此不能去推斷是藉由Apoptosis或是Autophagy的途徑來引發細胞死亡。綜合以上real-time PCR分析的結果，可以明顯看出桑葚對膀胱癌或是山葵、荖葉對肝癌細胞的凋亡途徑基因表現，未來可以著重在這些基因的研究。而這些研究可以提供有益的資訊在這些癌症的研究。

關鍵字

肝癌 ； 膀胱癌 ； 程式性死亡 ； 定量PCR陣列 ； 桑葚 ； 山葵 ； 荖葉 ； 基因表現)

並列摘要

貳.英文摘要
Bladder cancer and Liver cancer are difficult to detect at early stages, and are usually at the late stages when diagnosed. Defect in the regulation or execution of the programmed cell deaths (including apoptosis and autophagy) is one of the main causes of cancer. There are many genes participating in the apoptotic and autophagic programmed cell deaths. The molecular regulators of both pathways are inter-connected, and the cross-talk between them is quite complex. Previous studies have confirmed that Mours alba L. has antioxidant and anti-inflammatory effect, Wasabia japonica has anti-proliferative effect, and Piper betle has antioxidant and inhibitory effect on some cancers. In our laboratory, Mours alba L. extract has cytotoxic effect toward TSGH-8301 (bladder cancer cells), Wasabia japonica and Piper betle L. have cytotoxic effect toward Hep3B and HepG2 (liver cancer cells) as confirmed by MTT assay and flow cytometry. In this study real-time PCR analysis was used to observe the expression of apoptosis and autophagy related genes in the treated cells. The results showed that Mours alba L. extract induced apoptosis dependence genes FADD and caspase-3 in TSGH-8301 cell. The combined treatment of Mours alba L. extract and Taxol did not change the levels of the genes as compared to the Mours alba L. extract alone. Wasabia japonica extract induced apoptosis dependence genes FASLG and caspase-3 in Hep-G2. However, Wasabia japonica extract and Piper betle extract did not show significant effect of the examined genes in Hep3B. The outcomes of real-time PCR analysis clearly show that apoptotic pathway was induced in bladder cancer cells treated with Mours alba L. and in HepG2 liver cancer cells treated with Wasabia japonica. The could be the major focus in the future work.

並列關鍵字

Liver cancer ； Bladder cancer ； program death ； real-time PCR ； Mulberry ； Piper betle ； Gene expression

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