祕魯酸漿萃取化合物-AN37經由細胞凋亡以及氧化逆境路徑選擇性殺死口腔癌細胞

Selective killing of oral cancer cells by Physalis peruviana -derived AN37 compound through apoptosis and ROS pathway

黃若雯(Rou-Wen Huang)

指導教授 ： 張學偉

高雄醫學大學/生命科學院/生物醫學暨環境生物學研究所/碩士(2012年)

摘要

從祕魯酸漿萃取出來的化合物-AN37曾被證實有抗肝癌的效果，但是對於抗口腔癌的能力卻很少被研究。本篇研究想要評估AN37抗口腔癌能力以及選擇性殺死口腔癌細胞的能力。利用細胞生存率發現，AN37處理細胞24以及48小時後，口腔癌細胞Ca9-22的生存率曲線下降較口腔正常細胞human gingiva fibroblast-1 (HGF-1)快。分別依據propidine iodide/annexin V染色與??-H2AX染色的細胞流式分析，發現AN37誘導更多Ca9-22細胞凋亡，以及DNA雙股斷裂較HGF-1多。利用西方點墨法也發現細胞凋亡相關蛋白，如caspase-9、caspase-3、PARP1的表現，在Ca9-22有隨著濃度增加的趨勢，並且表現較HGF-1明顯。彗星實驗利用Formamidopyrimidine-DNA glycolyase (FPG)，也顯示AN37造成Ca9-22較多的DNA損傷(8-oxoG)，並且是由氧化誘導DNA受損。再利用細胞流速分析，發現AN37處理過後的Ca9-22產生較多的活性氧化物質(reactive oxygen species，ROS)；而粒線體膜電位較HGF-1降低更多。由這些結果顯示： AN37選擇性殺死口腔癌細胞是藉由細胞凋亡以及ROS相關路徑進行。

關鍵字

祕魯酸漿 ； 口腔癌 ； 細胞週期 ； 細胞凋亡 ； DNA雙股斷裂 ； 氧化活性物質 ； 粒線體膜電位

並列摘要

Physalis peruviana-extracted pure compound AN37 has reported to be anti-hepatoma, but the possible anti-oral cancer effect of AN37 is less addressed. Accordingly, the purpose of my study is to evaluate selective killing effect between oral cancer and normal cell using AN37. Previously, our laboratory found that AN37 has a selective killing ability in oral cancer cells Ca9-22 (gingival carcinoma) and normal oral cells HGF-1 (human normal gingiva fibroblast) using MTS assay. Using annexin V/propidine iodide (PI)-based and western blot apoptosis analysis indicated that AN37 induced apoptosis in Ca9-22 more than HGF-1. Using ??-H2AX-based fluorescence analysis, AN37 induced DNA double strand break in Ca9-22 more than HGF-1. It indicated that the DNA damage induced by ROS by comet assay and oxyDNA assay kit. Ca9-22 produced more reactive oxygen species (ROS) but less mitochondria membrane potential (MMP) than HGF-1 after AN37 treatment. It indicates that apoptosis and ROS-related pathways may be involved to selective killing of AN37 in oral cancer cells.

並列關鍵字

oral cancer cell ； selective cancer drug ； cell cycle ； reactive oxygen species ； mitochondria membrane potential

參考文獻

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