- 學位論文
雙硫侖對人類子宮內膜癌細胞生長及細胞週期之抑制作用探討
Exploring suppressed effect for proliferate and regulated cell cycle of the Tetraethylthiuram Disulfide treated to endometrial cancer cell
摘要
子宮內膜癌在全球女性癌症排名位居第五，目前在臨床治療子宮內膜癌的方式主要透過手術切除後，並合併與放射治療、化學治療、賀爾蒙治療進行術後的治療，前兩者在預後輔助效果較佳，但是對人體仍有較大的副作用。 因此開發對人體副作用較小的輔助治療藥物為先務之急。研究指出Tetraethylthiuram Disulfide (TETD) 具有抑制癌細胞生長、凋亡等反應，同時該藥物已被FDA (Food and Drug Administration)核准超過60餘年，並在乳癌細胞株MDA-MB-231及肺癌細胞株Huh1等，證實能抑制細胞增生與促進細胞凋亡等抑制癌細胞活性。 目前還沒有人利用TETD藥物對人類子宮內膜癌(人類子宮內膜癌第一型HEC-1A、人類子宮內膜癌第二型RL95-2)兩株細胞進行測試，因此我們利用TETD對 (人類子宮內膜癌第一型HEC-1A、人類子宮內膜癌第二型RL95-2)、人類腎臟上皮細胞(293T)進行測試，透過本研究探討TETD對於人類子宮內膜癌第一型HEC-1A、人類子宮內膜癌第二型RL95-2細胞株的影響與作用為何。 首先，在MTT結果顯示，當TETD濃度達0.1μM以上時，對人類子宮內膜癌第一型HEC-1A有明顯的生長抑制效果；但人類子宮內膜癌第二型RL95-2與293T並沒有觀察到明顯的生長抑制效果。利用流式細胞儀觀察細胞週期與細胞凋亡現象，在16小時會誘導人類子宮內膜癌第一型HEC-1A細胞週期延緩，48小時則有更加明顯的細胞週期延遲。而在蛋白分析中，發現TETD會抑制mutp53蛋白所活化的NF-κB/P65蛋白進而使細胞生長延緩。 目前已證實TETD對人類子宮內膜癌第一型HEC-1A具有生長抑制之效果，未來可以進行一步探討TETD抑制NF-κB與人類子宮內膜癌第一型HEC-1A之間的關係，以釐清在生長抑制之影響。
並列摘要
Endometrial cancer is the fifth most common cancer for female all over the world. For past decade, both incidence and mortality had been increased. So far, ablative surgery is a major therapy for endometrial patients on clinical which combined with chemotherapy, radiotherapy, hormone therapy to postoperative patients. The first two treatments have more efficacy for prognosis, but harmful. Thus, developing a lower toxic, more efficacy drug is necessary. According to previous studies, Tetraethylthiuram disulfide (TETD) shows anticancer activity and cell cycle arrest. In addition, TETD has been approved by FDA (Food and Drug Administration) it more than 60 years, and it is safe and useful under the consent dose for testing. In this study, we will explored the effect of Human endometrial cancer cell (HEC-1A, RL95-2) and Human Embryonic Kidney Cells 293 (293T) treated with TETD. Based on the result of cytotoxicity analysis, TETD (> 0.1μM) induced a significant cytotoxicity to HEC-1A, but non-effect for RL95-2 and lower hurting for 293T at TETD 0.3μM. The data of cytoflowmetry shows that, when TETD treated for 16 hours, it will induce the cell cycle delay, up to 48 hours has more significant of cell cycle delay with HEC-1A. Furthermore, we revealed that TETD selectively inhibits the NF-κB/P65 which activated by mutp53, leading to HEC-1A cell cycle delay. Although our data suggest that TETD specifical effect to cell cycle delay of HEC-1A, the mechanism of TETD is not clear, we will keep finding the pathway of TETD to endometrial cancer HEC-1A.