ATRA與Am80誘導HL-60分化的調控機制：微型核醣核酸與基因網路

Abstract

全反式維甲酸(all-trans retinoic acid, ATRA)與Am80是用於治療血癌，促使血癌細胞HL-60分化成顆粒球已經有很多年了，其中全反式維甲酸是天然的維他命A類似物，Am80則是經過合成改良的藥物，這兩種藥物對於誘導HL-60分化都有不錯的效果。在本篇的研究，我們利用基因微陣列晶片與生物資訊軟體的方法來挑選出在細胞分化中基因表現量上有差異的基因。軟體統計分析的結果顯示TGF-β訊號傳遞的生物路徑是ㄧ個被誘導出的生物路徑，就我們所知的，TGF-β訊號傳遞的生物路徑是ㄧ個與調控細胞分化、細胞增生有關的生物路徑，我們利用偵測與這個生物路徑有關的時間序列上的基因表現，來建構一個可能被ATRA與Am80所誘導的基因網路，再利用MiLINK這個軟體來找到這個可能與這個生物路徑有關的微型核醣核酸，這些微型核醣核酸可能扮演調控這個基因網路的角色，那微型核醣核酸他們主要是存在於非產生蛋白質的序列上，並且是單股約20~25個核甘酸的長度，他們被報導出具有調控細胞生長、細胞分化、還有一些疾病和癌症的產生有關，到現在為止，在人類基因組中已經有超過三百個微型核醣核酸被發現，在我們的結果中，miR-96與miR-107的表現都有顯著得被兩種藥物給誘導表現，這個結果告訴我們這個miR-96與miR-107可能是參與調控細胞分化的重要因子。

All-trans retinoic acid (ATRA) and Am80, natural and synthetic derivatives of Vitamin A, have been used in oncology for many years and are useful for inducing HL-60 to differentiate to granulocyte. In this study, we showed that TGF-β signaling pathway is the most perturbed and involved in differentiation process using microarray and bioinformatics approach. With time-series gene expression profiles measured by the quantitative real-time PCR, we constructed a gene network of ATRA- and Am80-induced human leukemia cell line HL-60 differentiation. The results from comparative analysis of target genes indicated that this gene network was regulated by some candidate microRNAs. MicroRNAs are non-coding, single-stranded RNAs of 20 ~ 25 nucloetides that execute a gene regulator function responsible for cell growth, cell differentiation, disease, and cancer. In our results, the expression of miR-96 and miR-107 were significantly induced by ATRA and Am80. In summary, our results showed that miR-96 and miR-107 seem to be the regulators involved in the gene network of leukemia cell differentiation.