Let-7f microRNA negatively regulates hepatic differentiation of human adipose tissue-derived stem cells
Let-7f microRNA negatively regulates hepatic differentiation of human adipose tissue-derived stem cells
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DOI：
10.1007/s13105-014-0346-z
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发表时间：
2014-09-01
影响因子：
3.4
通讯作者：
Arjmand, Sare
中科院分区：
文献类型：
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作者：
Davoodian, Nahid;Lotfi, Abbas S.;Arjmand, Sare
MicroRNAs(MiRNAs)是一类非编码RNA，除了调节细胞分化外，还参与调节新陈代谢、增殖和细胞周期等多种生物学过程。到目前为止，一些miRNAs已经被认为在调节肝功能方面具有重要作用。从统计学上讲，let-7f被认为是肝脏分化的负调控因子。在本研究中，我们研究了let-7f对人脂肪组织来源干细胞(HADSCs)肝脏分化的影响。用含人抑制物let-7f的重组慢病毒转导hADSCs，用实时定量聚合酶链式反应(qRT-PCR)检测肝细胞核因子α(HNF4a)、白蛋白(ALB)、甲胎蛋白(AFP)、细胞角蛋白18(CK18)和细胞角蛋白19(CK19)的表达。免疫细胞化学方法检测肝细胞标志物ALB、AFP、HNF4a的表达水平，并进行肝功能、糖原沉积、尿素分泌等生化分析。QRT-PCR显示LET-7f抑制剂慢病毒转导的细胞HNF4a、ALB、AFP、CK18和CK19的表达显著上调。免疫细胞化学检测ALB、AFP、HNF4a阳性染色。在处理的细胞中也发现了尿素的产生和糖原的沉积，这是肝细胞的两个特征。因此，let-7f沉默导致肝细胞特异性因子表达增加，促进了hADSCs的肝脏分化。综上所述，我们目前的报告提供了证据表明，抑制let-7f将有助于将hADSCs诱导为肝细胞样细胞，并可能在更广泛的肝病再生治疗中应用。
MicroRNAs (miRNAs) are noncoding RNAs involved in the regulation of the diverse biological processes such as metabolism, proliferation, and cell cycle, in addition to regulation of differentiation. So far, some miRNAs have been recognized to have important role in regulating hepatic functions. Statistically, let-7f has been revealed as a negative regulator of hepatic differentiation. In the present study, we investigated the effect of let-7f on hepatic differentiation of human adipose tissue-derived stem cells (hADSCs). hADSCs were transduced with recombinant lentivirus containing human inhibitor let-7 f. The expression of hepatocyte nuclear factors alpha (HNF4a), albumin (ALB), alpha fetoprotein (AFP), cytokeratin 18 (CK18), and cytokeratin 19 (CK19) was evaluated using quantitative real-time PCR (qRT-PCR). Immunocytochemistry was used to investigate the expression levels of the hepatocyte markers including ALB, AFP, and HNF4a, and biochemical analysis was implemented for hepatic function, glycogen deposition, and urea secretion. qRT-PCR showed significant upregulation in HNF4a, ALB, AFP, CK18, and CK19 expression in cells transduced with let-7f inhibitor lentiviruses. Moreover, positive staining was detected for ALB, AFP, and HNF4a using immunocytochemistry. Urea production and glycogen deposits were also found in the treated cells, the two specific features of the hepatic cells. Therefore, let-7f silencing led to the increased expression of the hepatocyte-specific factors and the accelerated hADSCs hepatic differentiation. Summing all these finding together, our present report has provided evidences that inhibition of let-7f would facilitate induction of hADSCs into hepatocyte-like cells and possibly in regenerative therapy of the liver disease in a wider spectrum.