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標題: HLJ1為新的caspase-3受質且其表現量可以促進非小細胞肺癌因UV所引起之細胞凋亡
HLJ1 is a novel caspase-3 substrate and its expression enhances UV-induced apoptosis in non-small cell lung carcinoma
作者: 林聖怡
Lin, Sheng-Yi
關鍵字: heat shock protein
lung cancer
UV irradiation
出版社: 生命科學系所
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摘要: 全世界癌症病患者,肺癌為主要的死亡原因之一。通常癌症是因為細胞的基因變異所引起,包含基因突變、異常的基因調控及致癌基因的過度表現,而腫瘤發生決定於癌細胞的生長與死亡速率。HLJ1屬於熱休克蛋40 (Hsp40)基因家族的一員,是一新的腫瘤抑制基因,與非小細胞肺癌病人的存活率及疾病復發率有顯著相關。然而HLJ1在細胞凋亡的角色至今尚未清楚。近來的研究認為熱休克蛋白可以調控細胞凋亡的發生,在本研究中,首先以不同內源性HLJ1蛋白質表現量的非小細胞肺癌細胞株照射UV輻射,之後以流式細胞儀分析。在七株肺癌細胞株中發現,細胞凋亡的比例與HLJ1的表現呈現正相關性。為了進一步了解HLJ1在細胞凋亡的角色,我們建立HLJ1轉殖細胞株,進行UV輻射的處理,發現在低HLJ1表現的CL1-5細胞株中大量表現HLJ1,會透過增強JNK及caspase-3的活化,最後促進UV輻射所導致的細胞凋亡增加。此外,UV輻射會降低凋亡的細胞內HLJ1表現程度。以廣泛性的caspase抑制劑zVAD-fmk及caspase-3專一性的抑制劑DEVD-fmk事先處理細胞,可以避免細胞受到UV輻射的傷害,減少細胞內HLJ1的下降。進一步的實驗,我們也發現位於HLJ1胺基酸第125-128的位置,具有一非典型的caspase-3辨認切位MEID。被切割的HLJ1會導致更多的細胞凋亡及抑制癌細胞貼附非依賴性的生長。此外,與野生型的HLJ1相比,caspase切位突變的HLJ1會抑制細胞凋亡及促進細胞的生長。由我們的研究結果證明在細胞凋亡的過程中HLJ1為caspase-3的受質,且與細胞凋亡的發生有正相關。
Lung cancer is the most common cause of cancer deaths in the world. Cancer is often caused by gene alterations including genetic mutations, aberration of epigenetic regulation and gene expression. Tumorigenesis is determined by the balance of cell proliferation and death rates. HLJ1, a member of the DnaJ-like Hsp40 family, is a newly identified tumor suppressor protein whose expression is highly associated with relapse and survival in non-small cell lung cancer (NSCLC) patients. However, the role of HLJ1 in apoptosis is still unknown. Recently, it has been demonstrated that heat shock proteins (HSPs) can regulate apoptosis. In this study, NSCLC cell lines with the varying HLJ1 expression levels were subjected to UV irradiation, followed by flow cytometry analysis. Interestingly, cell apoptosis in these cell lines were positively correlated with HLJ1 expression. To understand the role of HLJ1 during cell apoptosis, a human lung adenocarcinoma cell line with enforced expression of HLJ1 was established and exposed to UV irradiation. The enforcing expression of HLJ1 in low HLJ1-expressing cells promoted UV-induced apoptosis through enhancing JNK and caspase-3 activation. Additionally, UV irradiation led to the decrease of HLJ1 predominantly in apoptotic cells. The pan-caspase inhibitor, zVAD-fmk, and caspase-3-specific inhibitor, DEVD-fmk, prevented UV-induced degradation of HLJ1 by the late stage of apoptosis. Furthermore, we found a non-typical caspase-3 cleavage site (MEID) at amino acid 125-128 of HLJ1. Cleavage of HLJ1 led to more apoptosis and inhibited cancer cell anchorage independent growth. Moreover, the caspase-resistant mutant HLJ1 inhibited apoptosis and promoted tumor growth as compared to wild-type HLJ1. Take together, our results suggest that HLJ1 is a novel substrate of caspase-3 and the expression of HLJ1 is positively correlated with apoptosis.
其他識別: U0005-3007201022041600
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