Quantification of plasma hTERT DNA in hepatocellular carcinoma patients by quantitative fluorescent polymerase chain reaction

Abstract

Purpose: To investigate the levels of human telomerase reverse transcriptase (hTERT) DNA in the plasma of patients with hepatocellular carcinoma (HCC), and to evaluate the diagnostic value and correlation of hTERT DNA with clinical parameters in HCC. Methods: A real-time quantitative fluorescent polymerase chain reaction (FQ-PCR) system was designed and evaluated. Plasma samples were collected from 60 HCC patients, 21 patients with hepatitis B virus (HBV) and 29 healthy controls. Plasma DNA was extracted and quantified by FQ-PCR. The diagnostic value of plasma hTERT DNA levels and their relationships with clinical characteristics were analyzed statistically. Results: Plasma levels of hTERT DNA in HCC patients were significantly higher than in HBV patients (4.18×104±4.94×104 copies/μl vs 1.21×104±6.63×103 copies/μl, P=0.003) and healthy controls (4.18×104±4.94×104 copies/μl vs 1.44×104±6.61×103 copies/μl, P < 0.001). Receiver operating characteristic curve analysis indicated a sensitivity of 64% and a specificity of 90% for the ability of hTERT DNA levels to detect malignancy at a cutoff value of 1.87×104 copies/μl. Association analysis revealed that plasma hTERT DNA levels were closely related to tumor size, portal vein cancer embolus and TNM stage (P=0.013, P=0.010, and P=0.029, respectively), but were not associated with lymph node metastasis, hepatitis B surface antigen, or α-fetoprotein (AFP) (all P > 0.05). The levels of plasma hTERT DNA in HCC patients with AFP ≤20 ng/ml were significantly higher than in HBV patients (4.59×104±4.98×104 copies/μl vs 1.44×104±6.63×103 copies/μl, P=0.016) and in healthy controls (4.59×104±4.98×104 copies/μl vs 1.21×104±6.63×103 copies/μl, P=0.001). Conclusions: Quantitation of plasma hTERT DNA by FQ-PCR may provide a novel complementary tool with potential clinical applications for the screening and detection of HCC. Plasma hTERT DNA has the potential to be a broad tumor marker for common cancers.

Circulating Tumor DNA and Hepatocellular Carcinoma

“…44 While most studies evaluated the overall cfDNA concentration, one study evaluated the performance of wild-type human telomerase reverse transcriptase (hTERT) cfDNA from 60 HCC patients, 21 patients with hepatitis B virus (HBV), and 29 healthy controls. 45 Concentration of hTERT cfDNA had a sensitivity of 64% and a specificity of 90% to detect HCC. However, hTERT cfDNA levels were closely related to tumor size, portal vein cancer thrombus and TNM stage, limiting the independence of this marker from existing staging variables and biasing towards later stage disease.…”

Circulating Cell-Free DNA in Hepatocellular Carcinoma: Current Insights and Outlook

“…The authors confirmed their findings in a follow-up study with a larger cohort of 96 HCV-associated HCC patients and 100 HCV carriers without known HCC ( 44 ). Since then, studies have reported the concentration of cfDNA in serum or plasma to be 3–4 times higher in HCC patients compared to patients with chronic hepatitis ( 44 , 49 , 61 ) and nearly 20 times that of healthy controls ( 49 ). Several studies, including our own, have also reported that high cfDNA concentration was associated with larger tumors ( 40 , 41 , 58 ), higher tumor grade ( 40 , 58 ), shorter overall survival ( 41 , 44 ), and may serve as predictive biomarker for distant metastasis after curative surgery ( 44 ).…”

Correlation between the expression of hTERT gene and the clinicopathological characteristics of hepatocellular carcinoma

“…The activity levels of telomerase mainly rely upon the expression level of human telomerase reverse transcriptase (hTERT). In HCC patients, the activation of telomerase and a high expression level of the hTERT subunit catalyzed by telomerase have been reported ( 1 , 9 – 11 ). Furthermore, hTERT expression and telomerase activation has been identified in ≤90% of human malignant tumors, including gastric, brain and renal cancers ( 12 ).…”