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Abstract Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and is associated with liver cirrhosis (LC) in 80% of cases. miRNAs are approximately 22-nucleotide, noncoding, endogenous RNA molecules with an important role in a number of biological processes, including embryonic development, cell differentiation, and tumorigenesis. Recently, many studies suggest a link between aberrant expression of miRNAs and various cancers including HCC, recent studies have documented the involvement of miRNAs in HCC in tumor progression and metastasis. miRNA- 21 is one of the first miRNAs to be described as an oncomir as most of the targets of miRNA-21 are tumor suppressors such as Phosphatase and tensin homolog (PTEN), programed cell death 4 (PDCD4), Tropomyosin and others. It is upregulated in many types of human cancers. PDCD4 is a tumor suppressor gene that plays an important role in regulating apoptosis, invasion and metastasis.Several reports described the regulation of PDCD4 by miRNA-21. So this study aimed to evaluate the diagnostic potential of circulating miRNA-21 and study its effect on programed cell death 4 (PDCD4) gene expression in Egyptian patients with HCC. The present study was conducted by collaboration between Medical Biochemistry Department, Faculty of Medicine, Clinical Biochemistry and Oncology Departments, National Liver Institute, Menoufia University. It included 70 subjects: 30 patients with HCC, 20 patients with CLD secondary to HCV infection presented to the inpatient wards and outpatient clinic of Oncology Department, National Liver Institute, Menoufia University in the period from January 2014 to December 2014, and 20 unrelated apparently healthy subjects matching age and gender served as control group. All patients and control groups were subjected to the following: 1. Complete history taking. 2. Complete clinical examination. 3. Abdominal ultrasonography and/ or CT. 4. Laboratory investigations including: complete blood picture, liver function tests, viral markers (HBsAg, HCV-Ab), estimation of serum AFP and molecular testing for miRNA-21 expression and PDCD4 gene expression. The results of this study revealed the following: The groups were homogenous regarding age and gender. There was no significant difference between HCC and CLD groups regarding all studied variables (liver function tests and AFP) except ALP, GGT and AFP which showed significant increase in HCC group. There was significant decrease in hematological indices (Hb – RBCs – platelet count) in both HCC and CLD groups compared to control group. Meanwhile, no significant difference was detected between HCC and CLD groups. There was significant increase of miRNA-21 in HCC group compared to each of CLD and control groups. However, it showed no significant difference between CLD and control groups. PDCD4 gene expression showed significant decrease in HCC group compared to each of CLD and control groups. Meanwhile, it showed no significant difference between CLD and control groups. There was significant inverse correlation between miRNA-21and PDCD4 gene expression RQs in HCC group. There was significant positive correlation between miRNA-21 and AFP and significant inverse correlation between PDCD4 and each of total and direct bilirubin in HCC group. A significant positive correlation was detected between miRNA-21 and ALT as well as between PDCD4 gene expression and GGT in CLD group. Meanwhile, No significant correlation was detected between miRNA-21 and PDCD4 gene expression and all studied parameters in control group. The mean RQ of miRNA-21 showed significant increase, with presence of cirrhosis, increased number of focal lesions, larger size of tumor, advanced tumor stage and presence of vascular invasion. Meanwhile, the mean RQ of PDCD4 gene expression showed a significant decrease with increased number of focal lesions, larger sized tumors and and advanced stages of HCC. However, no significant difference in serum AFP was detected regarding the aforementioned variables. The mean RQ of circulating miRNA-21 and the mean serum level of AFP showed a highly significant elevation in HCC patients with early tumor development (single focal lesion, tumor size <3cm, and collectively TNM 1 and 2 stages) compared to each of the control and CLD patients groups, while no statistical difference was detected regarding the mean RQ of circulating PDCD4 gene expression. Receiver Operator of characteristics (ROC) curve analysis of AFP and miRNA-21: displayed that the best cut-off of serum AFP for differentiation of HCC cases from those without HCC was 91.7 ng/ml, at this cut-off; the sensitivity, specificity, PPV, NPV and overall accuracy were 75.2 %, 92.3 %, 90.2 %, 69.2%, and 77.0 % respectively. For miRNA-21 the best cutoff was 3.93 RQ with sensitivity, specificity, PPV, NPV and overall accuracy of 90 %, 94.8 %, 94.4 %, 87.5 % and 92.5 % respectively. Combined use of two parameters was superior to the use of AFP alone as the sensitivity, specificity, PPV, NPV and overall Accuracy were 91.3%, 97%, 96%, 90% and 92% respectively. |