Pyrogallol對人類非小細胞肺癌細胞增殖抑制效應和細胞程式死亡機制之探討

The Anti-Proliferative Effects and Apoptotic Mechanism of Pyrogallol on Human Non-Small Cell Lung Cancer Cell

陳煌麒(Huang-Chi Chen)

指導教授 ： 黃明賢

高雄醫學大學/醫學院/醫學研究所/碩士(2011年)

摘要

Pyrogallol 是從 Emblica officinalis 中萃取出來一種活性成分，在先前的研究中已被證實對許多人類的癌症細胞株有抗增殖與毒殺效用，且相對於 gallic acid， pyrogallol 對正常的支氣管表皮細胞株並無太大傷害。因此在本篇研究中，藉由探討細胞增生、細胞週期及細胞凋亡等機制，評估 pyrogallol 與傳統的化學藥物 taxol 或 cisplatin 合併使用，相對於taxol 或 cisplatin單獨使用時，對於人類肺癌細胞株 H441 與 H520 是否有加成作用。
在 MTT 法檢測結果中， pyrogallol 無論是與 taxol 或 cisplatin 合併使用都有明顯的加成作用。接著以流式細胞儀進行細胞週期分布的探討，同樣的合併使用後，停滯在 G2/M 期的細胞量也大幅上升。使用西方墨點法分析發現， cyclin B1、 Cdc2 與 Cdc25c 呈現下降趨勢，而磷酸化的 Cdc2 (Thr14) 則呈現上升趨勢。再者 PARP 的裂解增加也意味著肺癌細胞株的凋亡，同時以 Annexin-V 雙染與 TUNEL assay 均證實 pyrogallol 與 taxol 或 cisplatin 合併使用會造成更明顯的肺癌細胞株凋亡。由以上結果顯示， pyrogallol 在對於治療非小細胞肺癌上，可以與傳統的化療藥物同時使用以增加其臨床效果。

關鍵字

肺癌 ； pyrogallol ； 化學治療 ； 細胞週期 ； 細胞凋亡

並列摘要

Pyrogallol, a catechin compound, is an active component of Emblica officinalis extracts and has an antiproliferative effect on some human cancer cell lines. In our preliminary study, pyrogallol, not like the gallic acid, had highly cytotoxic effect on human lung cancer cell lines and less effect on human bronchial epithelium cell line. This study was performed to investigate the synergic effect of pyrogallol treated with taxol or cisplatin on human lung cancer cell lines – H441 (lung adenocarcinoma) and H520 (lung squamous cell carcinoma).
The MTT (cytotoxic) data showed significant growth inhibition of lung cancer cells followed pyrogallol treatment with taxol or cisplatin. The cell cycle of lung cancer cells was arrested in G2/M phase using flow cytometry. Using Western blot analysis, the cell cycle related proteins – cyclin B1, Cdc2 and Cdc25c were decreased and the phosphorylated Cdc2 (Thr14) was increased after pyrogallol treatment with taxol or cisplatin. Moreover, the higher cleavage of poly (ADP)-ribose polymerase (PARP) indicated that combined treatment resulted in apoptosis of lung cancer cells. The cell apoptosis was also directly demonstrated using Annexin V-FITC and TUNEL stain. These results suggest that pyrogallol can be developed as a promising anti-lung cancer drug combined with traditional chemotherapy particular for the non-small cell lung cancer (NSCLC).

並列關鍵字

lung cancer ； pyrogallol ； chemotherapy ； cell cycle ； apotopsis

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