探討使NUCB-2在肝癌細胞中過度表現後對癌細胞生長的影響

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Title:	探討使NUCB-2在肝癌細胞中過度表現後對癌細胞生長的影響 To investingate the effect of NUCB-2 overexpression on hepatocellular carcinoma cell
Authors:	李致翰
Contributors:	生物科技系洪瑞祥
Keywords:	肝癌內質網壓力 XBP-1 NUCB2 HCC ER stress XBP-1 NUCB2
Date:	2016
Issue Date:	2016-12-21 15:34:18 (UTC+8)
Abstract:	過去很多文獻都指出在很多的腫瘤組織都可以觀察到有內質網壓力(Endoplasmic reticulum stress, ER stress) 的現象，近年來有研究更進一步發現 ER stress 與癌症的生長、轉移、治療上的抵抗性有相當的關聯性。在我們先前的研究中發現肝癌細胞在內質網壓力下會誘導 Nucleobindin-2 (NUCB-2) 大量表現，然而 NUCB-2 的過量表現在肝癌細胞所扮演的角色仍然不清楚，所以本論文我們更深入的去探討 ER stress 下誘導 NUCB-2 的機制與其在肝癌細胞中所扮演的角色。結果顯示我們把 pNUCB-2-EGFP 質體送到Huh-7 細胞，可以明顯增加NUCB-2的表現。此外我們對NUCB-2 的 promoter進行分析，發現 NUCB-2 具有 ATF-6 以及 XBP-1 這兩個 ER stress下游所調控蛋白的結合位置，當利用XBP-1 shRNA降低肝癌細胞 Huh-7 中 XBP-1 的表現量後，利用即時聚合?連鎖反應 (Real-Time PCR) 來分析發現內質網壓力誘導劑 tunicamycin (TM) 所誘導 NUCB-2 的表現在 Huh-7 XBP-1 shRNA細胞中明顯的被抑制，未來我們會繼續分析在內質網壓力下誘導 NUCB-2 的機制和探討過量表現 NUCB-2 時對於化學治療的影響，期望能更深入的去探討對於腫瘤在未來化學治療時能更有效的治療策略。 Many studies have pointed out that in the past a lot of the literature of tumor tissue can be observed endoplasmic reticulum stress (Endoplasmic reticulum stress, ER stress) phenomenon. Recent studies have further found that tumor cell growth, metastasis and drug resistance are associated with ER stress. From our previous study, induction of nucleobindin-2 (NUCB-2) overexpressionwas mediated through ER stress in HCC cells, but expression mechanism of NUCB-2 is still unclear. Therefore, in this study, we want to investigate the mechanism and role of NUCB-2 expression in response to ER stress. The results have shown that we establish NUCB-2 expression by transfection of pNUCB-2-EGFP plasmid to Huh-7 cells. In addition, NUCB-2 promoter was analyzed, and found there are two ER stress associated transcription factors, ATF-6 and XBP-1, on NUCB-2 promoter. Down regulation of XBP-1 expression was performed by using XBP-1 shRNA in Huh-7 cells. Decreased NUCB-2 and GRP78 expression was observed in Huh-7 XBP-1 shRNA cells during ER stress by using Real-Time PCR analysis. In the future, we will continue to analyze and investigate the mechanism of induction of NUCB-2 under ER stress, and the relationship between NUCB-2 and chemical therapeutic drugs treatment. Consequently, this mechanistic research may provide a molecular basis to develop a novel cancer chemotherapeutic agent for high level expression of NUCB-2 in cancer patients.
Relation:	校外公開：2021-08-31,校內公開：2016-08-31，學年度：104，69頁
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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