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Functional Study of MicroRNA-7a Depletion Induced Zebrafish Early Onset of Non-alcoholic fatty liver disease
|Authors: ||Chang, Ching-He|
miR-7a;microRNA sponge;non-alcoholic fatty liver disease;zebrafish
|Issue Date: ||2018-08-22T06:31:09Z
|Abstract: ||肝癌(Hepatocellular carcinoma, HCC) 在癌症死因中位居第二。 HCC的主要病因為HBV、HCV的感染和酒精性肝病， 隨這生活型態的改變在臨床由非酒精性脂肪肝病(non-alcoholic fatty liver disease, NAFLD) 生成的HCC案例逐漸增多。 NAFLD並無明顯病徵，僅有輕微脂肪變性及肝細胞氣球樣變，隨後發展成非酒精性肝炎(non-alcoholic steatohepatitis , NASH)進一步肝硬化肝功能尚失， 最終走向HCC。 MicroRNA-7(miR-7) 為影響腫瘤的發生及發展的關鍵腫瘤抑制者， 先前研究指出ob/ob大鼠肝臟中發現有下調的現象; miR-7會經過標定結合進而調控PI3K/AKT/mTOR pathway已達到抑制腫瘤生成與轉移， Liu et al, (2016) 文獻證實此條訊息途徑和肝臟中的脂肪代謝有密切關聯。 在本篇研究中我們利用L-FABP promoter專一表現miR-7a-spong於斑馬魚肝臟， 偵測miR-7a目標基因CUL5、GSK3β、PIK3CD、mTOR、p70s6k表現量上升。 餵食實驗中高(20%) 及低(<5%)油脂飼養肝臟中表現miR-7a-sp的實驗室組相較於WT對照組肝臟中的油脂累皆積明顯較多，且在成魚切片後觀察到細胞型態的異常及嚴重脂肪肝趨勢，同時也偵測到了肝細胞增生的訊號與PPARα和C/EBPβ mRNA的表現量上升， 透過研究這些現象期望建立一可研究早發性NAFLD之斑馬魚品系，對肝臟疾病的預防及治療有所貢獻。|
MicroRNAs (miRNAs) are short non-coding RNAs of 20-24 nucleotides that play key roles in the regulation of gene expression. Recently, several researchers have reported that miRNA we links to the pathogenesis of non-alcoholic fatty liver disease (NAFLD) and the progression to non-alcoholic steatohepatitis (NASH). NASH may progress to advanced-stages of hepatic fibrosis, cirrhosis and liver failure, and then causes of death and hepatocellular carcinoma (HCC). However, there is rarely known about specific therapies for NASH. To development of new target therapy is urgently needed. MicroRNA-7 (miR-7) plays an important role in the occurrence and development of tumors as a key tumor suppressor. Previous research indicated that mir-7a is down-regulated in ob/ob mouse liver and HNF4a-NF-jB feedback circuit including mir-7 is identified in HCC, miR-7 directly inhibits a number of oncogenic targets and impedes cancer progression in vitro and in vivo. Therefore, we designed a miR-7a sponge expression vector drove by liver specific promoter L-FABP combined fluorescent gene to establish transgenic fish. We show that miR-7-silencing activity is specifically knocked down in the transgenic fish liver. As the insights into our research aims for studying NAFLD, feeding these transgenic fish with high fat diet (HFD) and low fat diet (LFD) . We observed the accrual of fat in hepatocytes with Oil Red O staining. Our data shows that miR-7a sponge transgenic fishs have more lipid droplet accumulation in liver than WT with HFD and LFD. In the future, we will plan to analyze miR-7 target genes and of cell proliferation and cell apoptosis in transgenic fish, it can be expected to activities help the study of NAFLD.
|Appears in Collections:||[生命科學暨生物科技學系] 博碩士論文|
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