| Hepatocellular carcinoma (HCC) is one of the most common malignancies, whose main features are onset occult, poor prognosis and high mortality. The early diagnosis of HCC is one of the important factors affecting the prognosis of patients with HCC. Until now, the detection means can not be effective for the early diagnosis of HCC. Thus, it is very important to find an early diagnostic marker for HCC. The increasing evidence has shown the deregulation of microRNA (miRNA) in HCC which may be involved in the pathological process of the development and metastasis of hepatocellular carcinoma (HCC). The development and progression of HCC is closely related to chronic viral hepatitis, liver cirrhosis and so on. It is significant to explore the role of miRNA in human hepatitis B-related hepatocellular carcinoma.AIM:Increasing evidence has shown that the deregulation of microRNAs (miRNAs) is closely related to the development and progression of hepatocellular carcinoma (HCC). To screen for HCC-specific miRNAs, this study investigated the differentially expressed miRNAs between HCC and matched non-tumorous tissue (NT).METHODS:The cancer tissues and their corresponding adjacent tissues from the11patients with HCC were selected, and then the total RNA (containing miRNA) of each sample were extracted. The appropriate amount of RNA sample from the total RNA solution of11cases of HCC tissues constituted the HCC RNA pool (Tmix), and the RNA solution of matched NT tissues also constituted the NT RNA pool (NTmix) in a similar manner. This study analyzed the differential expression profiles of miRNAs in11pairs of HCC and matched NT from11hepatitis B virus (HBV) infection patients with the RT2miRNA PCR array containing88human cancer-related miRNAs. The fold change value was more than two between the HCC and the matched NT, which indicated that there was deregulation of miRNAs. The fold-change values of the miRNAs calculated by Tmix/NTmix were greater than2, which indicated the presence of deregulation. The fold-change values greater than2indicate up-regulation, and fold-change values less than0.5indicate down-regulation. The down-regulated let-7a was validated in another sample set of34tissues with the TaqMan RT-qPCR method.RESULTS:Combined with the amplification and melting curves, the results of the triplicate experiments were comprehensively analyzed. Nine miRNAs out of88human cancer-related miRNAs were excluded due to poor quality amplification. The remaining79miRNA were used to analysis the miRNA expression profiling of between the HCC and the matched NT tissues. Compared with the matched NT tissues,9miRNAs were up-regulated in the HCC tissues, and three were considered statistically significant (p<0.05):miR-96, miR-183, and miR-196a, which were up-regulated4.746-,7.127-, and3.498-fold, respectively. Simultaneously,9miRNAs were down-regulated in the HCC tissues, and two were considered statistically significant:let-7c and miR-138, which were down-regulated3.945-and4.790-fold, respectively. The expression levels of let-7a were1.071±0.401,0.926±0.477,0.881±1.214, and0.535±0.719in the healthy group, chronic hepatitis B(CHB) group, NT group, and HCC group, respectively (p>0.05).CONCLUSIONS:This study demonstrates that18miRNAs were deregulated in the HCC and matched NT tissues. The deregulated miRNAs suggest that further analyses with larger miRNA samples as a diagnostic marker are warranted. The gradually downward trend of Let-7a suggestes that let-7a may be involved in the evolution of HCC. |
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