Tolemerase Reverse Transcriptase Gene Expression as a Tumor Marker for Hepatocellular Carcinoma

Abstract

Problem statement: Hepatocellular carcinoma will emerge as a major form of malignancy in the coming decades. The continuing high incidence of hepatocellular carcinoma, suggests that this disease will continue to represent a global health problem far into the future. Different genes encode for the various components of the human telomerase complex. These components include the human Telomerase RNA Component (hTERC) and the Telomerase Catalytic Subunit (hTERT). Correlation between Telomerase Reverse Transcriptase (hTERT) expression and telomerase activity has been reported in cancer patients. This work aimed to clarify the significance of human Telomerase Reverse Transcriptase (hTERT mRNA) as a potential molecular tumor marker for Hepatocellular Carcinoma (HCC). Approach: The current study included 25 patients of hepatocellular carcinoma (HCC), 30 patients with liver cirrhosis and 25 age and sex matched individuals with normal laboratory and Image findings as a control group. hTERT mRNA was measured in plasma by Real time PCR in all patients samples in comparison with normal healthy controls. Results: The expression of hTERT mRNA by relative unit was 129.10±27.6 with range (67.72-69.6) Vs 5245.87±2382.48 (2053-12232.90) Vs 92782.76±16158 (61783.25-118596.47) for control Vs cirrhosis Vs HCC group respectively. The hTERT expression was significantly with 699 and 33 fold increase in HCC and cirrhosis groups correspondingly when compared to that of controls p<0.05. Conclusion: It was suggested that this procedure was highly discriminating between healthy subjects and cancer patients and strongly support the idea that a valuable diagnostic test for cancer might be developed using this genetic marker in plasma. However it needs to be combined with other markers in future studies to be more specific for liver cancer.

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“…Relative quantification of TERT mRNA and caspase-8 mRNA gene by real time PCR using SYBR green. 19,20 2.3.1. RNA extraction Total RNA was extracted using Gene JETä RNA Purification Kit (Fermentas) following the manufacturer's instructions and the standard protocol.…”

“…For each sample, 5 ll cDNA was used for RT-PCR using the Power SYBRÒ Green RT-PCR Master Mix kit (Applied Biosystems, Foster City, CA, USA) and the real time machine (Applied Biosystems one step, Foster City, CA, USA) to detect relative expression levels of mTERT mRNA 19 and caspase-8 mRNA. 20 The reaction conditions used to detect TERT and caspase-8 levels were: initial DNA denaturation at 95°C for 10 min and then 40 cycles of denaturation at 95°C for 15 s, annealing and elongation at 60°C for 1 min.…”