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標題: Imiquimod對非皮膚癌細胞株之作用探討
The Effect of Imiquimod on Non-skin Tumor Cell Lines
作者: 蕭安恬
Hsiao, An-Tien
關鍵字: Imiquimod;非皮膚癌細胞株
出版社: 醫學科技研究所
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Imiquimod(1-isobutyl-1H-imidazo[4,5-c]quinolin-4-amine),為一種免疫反應調節劑,能透過和免疫細胞上的Toll-like receptor 7的結合,啟動受體下游訊息傳遞反應,誘使免疫細胞分泌細胞激素,達到抑制病毒或腫瘤的效果。在臨床上已被廣泛用於治療菜花(尖銳濕疣,生殖器疣,性病疣),以及治療淺表型基底細胞癌和光化性角化病,是目前皮膚癌主要局部外用藥。除了調節免疫的作用之外,也有研究指出,imiquimod也會造成黑色素細胞癌的粒線體外膜通透性增加,釋放細胞色素c活化下游的caspase 9、caspase 3 直接誘導黑色素細胞癌進行細胞凋亡。本實驗即是利用AGS、HepG2、A549、T24等非皮膚腫瘤細胞,觀察imiquimod對其產生的反應。本研究證實imiquimod會直接誘導AGS、HepG2、A549進行細胞凋亡,同時發現細胞凋亡並不是imiquimod誘導腫瘤細胞死亡的唯一路徑。我們在實驗中發現imiquimod也會誘導細胞走向細胞自噬。細胞自噬的產生是當細胞遭受壓力時,細胞進行求生的路徑,但若當細胞透過細胞自噬仍無法存活時,則細胞便會走向第二型的細胞死亡。在我們的實驗裡,受imiquimod刺激的A549和T24細胞中均可觀察到細胞在進行細胞自噬時的重要特徵,LC3-I蛋白質轉換變成LC3-II,以及細胞質中有酸性自噬囊泡的產生。綜合這些結果得知,imiquimod會誘導AGS、HepG2細胞進行細胞凋亡。A549細胞經過imiquimod刺激後,會同時有細胞凋亡及細胞自噬的情形產生。另外imiquimod對於T24細胞的影響則是偏向誘導細胞進行細胞自噬的路徑。證實imiquimod作用於不同的腫瘤細胞株上,會誘導不同的作用機轉。並且在imiquimod的刺激下,細胞自噬對腫瘤細胞而言可能為一個抵抗細胞凋亡的保護機制。

Imiquimod(1-isobutyl-1H-imidazo[4,5-c]quinolin-4-amine), an immune response modifier, has potent antitumor activity. Imiquimod induces local production of cytokines through the interaction with Toll-like receptor 7 on immune cells. It shows clinical efficacy against several benign and malignant skin lesions, including actinic keratosis and basal cell carcinoma. In addition to the effects of immune modulation, imiquimod had been reported to possess considerable proapopotic activity against melanoma cells, mediated through Bcl-2-dependent release mitochondrial cytochrome c and subsequent activation of caspase 3 and caspase 9. In this study, we examined the imiquimod-induced cell death in different cancer cell lines. We discovered that imiquimod induced cell apoptosis and decreased the level of anti-apoptotic proteins in AGS and HepG2 cells. However, the percentage of sub-G1 cells showed no significant difference in A549 and T24 cells after imiquimod treatment. We show here that apoptosis is not the only mechanism of cell death caused by imiquimod. Autophagy may be a mechanism of stress-adaption by which the cell is trying to survive. We observed that imiquimod induce autophagy, indicated by LC3-I converted to LC3-II and developed cytoplasmic acridic orange (AO) AO puncta in A549 and T24 cells. In addition, imiquimod induced caspase 3 activetion in A549. Taken together, we observed that imiquimod induce both autophagy and apoptosis in A549 cells. However, only autophagy is significantly increased in the T24 upon imiquimod insults. Thus, the imiquimod may have differential effects on different cancer cell lines, and autophagy may be an important protective mechanism in specific cancer cells which have the ability to resistant imiquimod-induced apoptosis.
其他識別: U0005-1708200918295000
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