| 摘要: | 根據全球統計，膀胱癌在常見癌症中位居第十，而其中更好發於男性，常見的症狀如血尿、排尿時疼痛等狀況。臨床上對於肌肉浸潤型膀胱癌的治療多進行膀胱切除術，但除了患者在使用其他排尿路徑上的不便，也會影響術後的存活率，保存膀胱下的輔助治療方式（如：細胞凋亡療法）也成為找尋的目標。
細胞凋亡是目前常用於研究治療癌症的方法，根據研究顯示，使用血吸蟲可溶性蟲卵抗原 (Soluble Egg Antigen, SEA)作用的細胞，會經由內在或外在路徑影響caspase-3的表現，進而活化下游的凋亡路徑。細胞焦亡則是一種發炎性細胞死亡。Gasdermin D (GSDMD)是細胞焦亡的執行者，藉由caspase-1切割後，產生具有活性的N-GSDMD，進而與細胞膜上的磷脂蛋白結合形成孔洞，釋放被caspase-1活化切割形成的IL-1β和I L-18，造成發炎反應。細胞焦亡會發揮腫瘤抑制功能並且引起抗腫瘤免疫反應。
因此本論文以人類膀胱癌細胞 (HTB-9)和正常人類膀胱上皮細胞 (SV-HUC-1)作為細胞研究模式，經曼森血吸蟲 (Schistosoma mansoni)的可溶性蟲卵抗原蛋白 (SEA)作用後，我們發現膀胱癌細胞 (HTB-9) 比正常膀胱上皮細胞 (SV-HUC-1)更易發生凋亡及焦亡反應，特別是在濃度12和16 ?g/mL時效果更顯著。西方墨點法數據顯示，caspase-3、caspase-8、caspase-9 和 caspase-1 的表現量在 SEA的作用下，HTB-9和SV-HUC-1中有顯著差異性，HTB-9的表現都比SV-HUC-1高出許多。我們利用流式細胞儀檢測發現膀胱癌細胞中的caspase-1及caspase-3平均螢光細胞數值顯著增高，且在同濃度下細胞焦亡比例高於凋亡。顯示SEA對 HTB-9的誘導與發炎路徑活化焦亡反應有較大關係。然而，本研究仍需要更多數據來釐清SEA在促進癌細胞凋亡和焦亡中的詳細作用機制，以及其開發作為輔助藥物與現有膀胱癌治療方法配合使用的可能性。
Bladder cancer is the tenth most common cancer worldwide and more common in men. Symptoms include hematuria, pain when urinating and other conditions. Cystectomy is usually used to treat muscle-invasive bladder cancer, however, in addition to the inconvenience from the patients using other urination routes, the postoperative survival rate will also be affected. Therefore, there is an increasing focus on bladder-preserving adjuvant treatments, such as apoptosis-based therapies.
Apoptosis is currently common course in cancer treatment. Cells treated with Schistosoma soluble egg antigen (SEA) will affect the expression of caspase-3 through intrinsic or extrinsic pathways, and then activate the downstream apoptotic pathway. Pyroptosis is an inflammatory cell death, which caspase-1 and gasdermin D (GSDMD) are the key points of pyroptosis. After being cleaved by caspase-1, active N-GSDMD will combine with phospholipid protein on the cell membrane to form a hole, causing an inflammatory pyroptosis. Pyroptosis acts as a tumor suppressor and could elicit antitumor immune responses.
In this study, human bladder cancer cell line (HTB-9) and normal human bladder cell line (SV-HUC-1) were used as models. After treatment with Schistosoma mansoni soluble egg antigen protein (SEA), we found that bladder cancer cells (HTB-9) are more susceptible to apoptosis and pyroptosis than normal bladder epithelial cells (SV-HUC-1), especially at concentrations of 12 and 16 ?g/mL. Western blotting data showed that the expression levels of apoptotic caspase-3, caspase-8, caspase-9 and pyroptotic caspase-1 were significantly different between HTB-9 and SV-HUC-1 with SEA treatment. Their expression in HTB-9 was much higher than that in SV-HUC-1. Using flow cytometry, we further found that both the average fluorescent cell numbers of caspase-1 and caspase-3 in bladder cancer cells were significantly increased, and the proportion of cell pyroptosis was higher than apoptosis at the same concentration.
It is shown that the cytotoxicity induction of bladder cancer cells by SEA is closely related to the activation of pyroptosis through the inflammatory pathway. However, more data is still needed to clarify the detailed mechanism of effects of SEA in promoting apoptosis and pyroptosis in bladder cancer cells, as well as the possibility of its development as an adjuvant drug for use with existing bladder cancer treatments. |
