الفهرس 
Hepatocellular CarcinomaOnly 14 pages are availabe for public view
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Abstract
Evaluation of circular RNA network expression in Hepatocellular carcinoma:
This study was done at Medical Biochemistry Department, Ain Shams Faculty of Medicine during the period from 2015– 2018 and included 102 patients and 18 normal volunteers.
The aim of the present study was to evaluate hsa_circ_002136, miRNA-106b and mRNA-AAK1 network usefulness as serum molecular marker for HCC early detection. hsa_circ_002136, miRNA-106b and mRNA-AAK1 levels were measured using real-time polymerase chain reaction (qPCR) in serum. The studied groups:
group I: including 60 HCC patients
group II: including 42 HCV patients.
group III: including 18 healthy volunteers.
All patients in our study were subjected to full personal history taking, general and local examination routine laboratory investigations including: (Complete blood picture, Liver profile: serum albumin, serum alkaline phosphatase and prothrombin time (PT). Serum AST, ALT, Alpha fetoprotein, bilirubin and PCR for HCV and Renal function tests), radio diagnostic investigations as Abdominal ultrasound and Triphasic spiral CT.
Serum sample was taken for exosomal RNA extraction and real-time polymerase chain reaction (qPCR) for measuring mRNA-AAK1, hsa_circ_002136 and miRNA-106b level. The data were normalized using the endogenous ACTB and RNU U6 as reference control. The threshold cycle (Ct) value of each sample was calculated with Step One PlusTM software v2.2.2 (Applied Biosystems), and the 2- ΔΔCT method was used in the analysis of PCR data for relative quantification of mRNA-AAK1, hsa_circ_002136 and miRNA-106b.
The results were calculated and statistically analyzed by the SPSS software.
Participants were excluded from the study if Patients actively undergoing chemotherapy or radiation therapy for HCC or patients diagnosed with other malignancies or treated within the last 5 years.
group I: (Malignant)
mRNA-AAK1 qPCR: the percentage of patients who were mRNA-AAK1 positive in this group was 91.7%.
hsa_circ_002136 qPCR: the percentage of patients who were hsa_circ_002136 positive in this group was 90%.
miRNA-106b qPCR: the percentage of patients who were miRNA-106b positive in this group was 100%.
group II: (HCV)
mRNA-AAK1 qPCR: the percentage of patients who were mRNA-AAK1 positive in this group was 57.1%.
hsa_circ_002136 qPCR: the percentage of patients who were hsa_circ_002136 positive in this group was 2.4%.
miRNA-106b qPCR: the percentage of patients who were miRNA-106b positive in this group was 28.6%.
group III (Healthy normal)
mRNA-AAK1 qPCR: the percentage of patients who were mRNA-AAK1 positive in this group was 0%.
hsa_circ_002136 qPCR: the percentage of patients who were hsa_circ_002136 positive in this group was 0%.
miRNA-106b qPCR: the percentage of patients who were miRNA-106b positive in this group was 0%.
A significant difference was observed in Positivity rate of serum mRNA-AAK1, hsa_circ_002136 and miRNA-106b levels (RQ) in relation to all groups (p < 0.001).
The overall AFP sensitivity was 90%, specificity was 85%, PPV was 85.7%, NPV was 87.5% and accuracy was 87.5%. The positivity rate of AFP in different groups of this study showed a high significant increase in malignant group (90%) as compared with HCV group (21.4 %) and healthy control group (0%), (p= 0.000).
Applying (≤ 15.01), as a cut off value which was calculated by ROC curve, serum mRNA-AAK1 as a tumor marker showed 91.6% sensitivity, 63.6% specificity, 69.6% PPV, 83.7% NPV and 75.8% accuracy.
Applying ( ≤ 0.596), as a cut off value which was calculated by ROC curve, serum hsa_circ_002136 level using the RQ values 90% sensitivity, 98.3% specificity, 98.2% PPV, 90.8% NPV and 94.1% accuracy.
Applying (≥ 1.73), as a cut off value which was calculated by ROC curve, serum miRNA-106b level using the RQ values 100% sensitivity, 80% specificity, 83.3% PPV, 100% NPV and 90% accuracy.
There was no significant difference observed in differential expression of serum mRNA-AAK1, hsa_ circ_002136 and miRNA-106b (RQ) levels among malignant group in relation to mean age, sex, smoking, HCV-Ab, HBV-sAg, Cirrhosis, Child-Pugh score, BCLC stage and Average Tumor size (p> 0.05).
A significant difference was found between mean rank of serum mRNA-AAK1, hsa_circ_002136 and miRNA-106b (RQ) levels in malignant group as (41.09), (33.67), (83.25) compared with either HCV group (68.64), (85.83), (39.14) or healthy normal group (106.19), (90.83), (34.5) respectively (p<0.001).
On combining the Investigated Biomarkers among all groups of the study; the sensitivity , specificity , positive predictive value , negative predictive value and accuracy for serum hsa_circ_002136, miRNA-106b and mRNA- AAK1 gene expression were found to be (100% , 63.2%, 41.7%, 100%, 70.8%) respectively. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy for serum hsa_circ_002136, miRNA-106b, mRNA-AAK1 gene expression and serum AFP are (100%, 63.2%, 41.7%, 100%, 70.8%) respectively.
In early stage of Hepatocellular carcinoma the sensitivity, specificity, positive predictive value, negative predictive value and accuracy for serum miRNA-106b, hsa_circ_002136 gene expression and serum AFP are (100%, 88.6%, 90%, 100%, 84.1%) respectively.