Nguyen Bao Quoc*, Nguyen Doan Nguyen Phuong, Tang Kim Ngan, Nguyen Thi Minh Linh, Pham Hung Cuong and Nguyen Ngoc Bao Chau Pages 92 - 99 ( 8 )
Background: Hepatocellular carcinoma (HCC) is the leading cause of cancer-related death in the world and considered as one of the most susceptible cancers in humans. The microRNA molecule, hsa-miR122, considered as a potential biological marker linked with the injury of hepatocellular tissue, is the most common microRNA in human liver cancer. Understanding the expression profile of hsa-miR122 plays an important role in the diagnosis of HCC.
Objective: Identification and comparison of cut-off values of plasma hsa-miR122 expression were conducted in blood samples of healthy control, HBV infected and HBV-related HCC Vietnamese patients.
Methods and Result: Fifty-two blood samples of healthy control and HBV-related HCC cases, collected between 2015 and 2017 were obtained from Ho Chi Minh City Oncology Hospital, Vietnam. Written informed consent was attained from all patients and the Human Research Ethics Committee, Oncology Hospital (#08/BVUB-HDDD) approved the research protocol. Total RNA was isolated from blood samples with TrizolTM Reagent (Thermo Fisher Scientific, USA). To analyze the expression level of hsa-miR122, miRNA specific reverse transcription was performed using Sensi- FASTTM cDNA Synthesis Kit (Bioline, UK) as described by the manufacturer, followed by running RT-qPCR with SensiFASTTMSYBR No-ROX Kit (Bioline, UK). The housekeeping gene, GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used for normalization. The presence of hsamiR122 and HBV-DNA was identified in human blood using RT-PCR and LAMP techniques. Downregulation of plasma hsa-miR122 was observed in HBV-related HCC patients with a Ct value of 7.9 ± 2.1 which was significantly lower than found in healthy control (p<0.01). The loss of hsa-miR122 expression was observed in HBV infected patients. We also identified the difference of diagnostic values of this microRNA in different populations and provided a high diagnostic accuracy of HCC (AUC = 0.984 with sensitivity and specificity of 96% and 94%, respectively).
Conclusion: hsa-miR122 was downregulated in HBV-related HCC patients and found to be lower by approximately 10 fold than in healthy control, resulting in a potential biomarker for microRNA based diagnosis of HCC in human blood.
HBV, HCC, hsa-miR122, human blood, RT-qPCR, biomarker.
Research Institute of Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Research Institute of Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Research Institute of Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Ho Chi Minh City Oncology Hospital, Ho Chi Minh City, Ho Chi Minh City Oncology Hospital, Ho Chi Minh City, Faculty of Biotechnology, Ho Chi Minh City Open University, Ho Chi Minh City