水線草之光動力療法在引起人類皮膚癌細胞凋亡過程中造成細胞骨架受損機制之探討;Investigation of the mechanisms of cytoskeletal damage during photodynamic therapy with Hedyotis corymbosa-induced human skin cancer cells apoptosis

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Title:	水線草之光動力療法在引起人類皮膚癌細胞凋亡過程中造成細胞骨架受損機制之探討;Investigation of the mechanisms of cytoskeletal damage during photodynamic therapy with Hedyotis corymbosa-induced human skin cancer cells apoptosis
Authors:	陳美宇;Mei-Yu Chen
Contributors:	中國醫藥大學：中國藥學研究所
Keywords:	光動力療法;水線草;M21皮膚癌細胞;細胞凋亡;細胞凋亡相關蛋白;細胞骨架;粒線體;Photodynamic therapy;Hedyotis corymbosa;M21 skin cancer cell line;apoptosis;apoptosis-related proteins;cytoskeleton;mitochondria
Date:	2008-07-18
Issue Date:	2009-08-10 18:02:14 (UTC+8)
Abstract:	水線草是茜草科植物，為常用且具抗癌效果的民間醫療用藥。本實驗主要的目的是探討水線草酒精粗抽物配合光動力療法引發M21皮膚癌細胞凋亡的作用及機制。光動力療法已被證實對惡性腫瘤有很好的功效。本研究指出，處理80 μg/ml水線草酒精粗抽物配合0.8 J/cm2光照劑量可以誘發M21細胞死亡，水線草酒精粗抽物配合光照引發M21細胞死亡是一種典型的細胞凋亡，在此細胞凋亡的過程中伴隨著凋亡小體的產生、染色質縮合及磷脂醯絲胺酸外翻等現象。本實驗結果也顯示，Bcl-2與caspase家族等與細胞凋亡相關蛋白、粒線體通透性轉運孔的開啟、粒線體膜電位下降及粒線體活性降低等因素都參與在水線草酒精粗抽物配合光動力療法引發M21細胞凋亡之途徑中。我們也證實水線草酒精粗抽物配合光照引發M21細胞中組成細胞骨架之肌動蛋白絲及微小管蛋白的改變。細胞骨架穩定劑phalloidin被證實可部份抑制水線草酒精粗抽物配合光照引發M21細胞之細胞骨架及粒線體膜電位的改變。綜合以上結果證實水線草酒精粗抽物配合光照引發M21細胞凋亡的途徑中，水線草酒精粗抽物配合光照對於細胞骨架的影響應在粒線體功能的上游。我們也證實內質網壓力也參與在水線草酒精粗抽物配合光照引發M21皮膚癌細胞凋亡的途徑中。 Hedyotis corymbosa (L.) LAMARCK (Rubiaceae) has been used in traditional Chinese medicine and found to have anti-tumor effect. The major purpose of this study was to characterize the effects and mechanisms of apoptosis induced by photodynamic therapy (PDT) with Hedyotis corymbosa extracts in M21 skin cancer cells. PDT was found to be an effective therapy for local malignant tumors. Our study indicated by 0.8 J/cm2 fluence dose caused cells death. PDT with H. corymbosa extracts-induced M21 cells death is a typical apoptosis that was accompanied by formation of apoptotic bodies, nuclear condensation and externalization of phosphatidylserine. In this study, PDT with H. corymbosa extracts-induced apoptosis of M21 cells involved modulation of the expression of apoptosis-related proteins, such as Bcl-2 and caspase family members, and mitochondrial function, such as opening mitochondrial permeability transition pore, disruption of mitochondrial membrane potential and the decrease in mitochondria uptake activity. We also demonstrated that cytoskeleton is the target of PDT with H. corymbosa extracts-induced M21 cells apoptosis. The change of distribution of F-actin was partly inhibited by pretreatment with phalloidin, an actin-stabilizing agent. PDT with H. corymbosa extracts-induced the disruption of mitochondrial membrane potential was also partly blocked by pretreatment with phalloidin. It seems to indicate that cytoskeleton lies upstream of mitochondria in PDT with H. corymbosa extracts-induced M21 cells apoptosis. Furthermore, we have demonstrated that the endoplasmic reticulum stress was involved in H. corymbosa extracts-induced apoptosis in M21 skin cancer cells.
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