- 學位論文
慢性C型肝炎患者周邊白血球中microRNA的表現與疾病發展的相關性
The association of microRNA expression in peripheral blood lymphocytes and disease progression of chronic hepatitis C patients
摘要
研究目的：C型肝炎病毒 ( Hepatitis C virus ) 感染是造成肝癌的重要原因之一。本研究想了解在C型肝炎罹病及治療過程中微型核醣核酸 ( microRNA ) 可能產生之影響及變化，並探討其變化與治療療效間之相關性，希望能對C型肝炎臨床治療上有所幫助。另外也可望透過microRNA之篩選，對C型肝炎病患之診斷及再復發的可能性能提出新的判斷依據。 研究方法及資料：首先挑選C型肝炎患者與未受C型肝炎感染者 ( 對照組 ) 各10位之血液檢體做microRNA表現輿圖分析 ( microRNA microarray analysis ) 並由分析結果和已知文獻中選出與對照組表現相比有明顯差異之microRNAs及其下游基因利用及時定量聚合酶連鎖反應 ( Real-time RT-PCR ) 做進一步研究。 研究結果：在慢性C型肝炎患者周邊白血球中，miR-16、miR-193b、miR-199a-3p、miR-222及miR-324-3p表現量增加，而miR-122表現量減少。經過長效型干擾素 ( IFN-alfa ) 與雷巴威林 ( Ribavirin ) 治療後，只有miR-222表現量有增加現象，而miR-16、miR-122、miR-193b及miR-199a-3p表現量減少。此外，miR-324-3p在HCV基因型第一型 ( GT-1 ) 或治療失敗 ( non-SVR ) 患者表現量增加，但HCV基因型非第一型 ( GT-non1 ) 患者表現量卻減少。然而，結束治療三至六個月後，microRNAs表現都有增加之情形。再進一步分析其下游調控基因，發現Mcl-1與miR-193b、miR-199a-3p及miR-324-3p有顯著負相關情形，而Cyclin D1僅和miR-16、miR-324-3p有明顯負相關的現象。 結論與建議： 本研究未發現microRNA與藥物治療效果 ( SVR ) 有直接關係，但其中miR-193b與 miR-199a-3p有高度鑑別慢性C型肝炎之能力。且C型肝炎治療後的實驗結果推測microRNA表現可能與免疫調節有關，需要未來長期及深入研究。
並列摘要
Objective：Infection with the hepatitis C virus is one of the risk factors in the development of liver cancer. This aim of this study is to understand the effect of microRNA expression levels on hepatitis C development and treatment response, and to provide evidence that the expression levels of microRNA can be used as biomarkers for the diagnosis and assessment of recurrence of hepatitis C. Materials and Methods：First, peripheral blood lymphocytes (PBLs) were collected from 10 hepatitis C patients and 10 health controls for microRNA profile analysis. Based on the resulting microarray data and published references, 7 candidate microRNAs were selected, and real-time RT-PCR was used to further confirm the microRNAs and target genes expression in patient’s PBLs. In addition, the down-stream genes Mcl-1 and Cyclin D1 were also analyzed through real-time RT-PCR. Results：In chronic hepatitis C patients’ PBLs, miR-16, miR-193b, miR-199a-3p, miR-222 , and miR-324-3p expression levels were increased and miR-122 expression was decreased, in comparison with the health control group. After IFN-alfa and Ribavirin treatment, only miR-222 expression levels were decreased, while miR-16, miR-122, miR-193b and miR-199a-3p expression were increased. In addition, the expression levels of miR-324-3p in HCV genotype-1 and non-SVR patients were increased, but were decreased in the expression levels of HCV genotype-non1 patients, when compared with non-treatment. Three to six months after the end of treatment, the expression of these microRNAs were increased. Further analysis of the downstream genes Mcl-1and cyclin D1 of these microRNA showed that mcl-1 was negatively correlated with miR-193b, miR-199a-3p, and miR-324-3p expression, while Cyclin D1 was only negatively correlated to miR-16 and miR-324-3p. Conclusion and Suggestions：This study did not find a direct correlation between microRNA expression and the effects of IFN-alfa and Ribavirin treatment in patients with hepatitis C, but miR-193b miR-199a-3p showed high specifics for hepatitis C diagnosis. It is suggested that changes in microRNA expression may correlate with immune regulation, but this requires further study.
參考文獻
延伸閱讀
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