Analysis of alpha-fetoprotein mRNA in peripheral blood: detection and semi-quantitation by reverse transcription polymerase chain reaction

Citation
W. Luo et al., Analysis of alpha-fetoprotein mRNA in peripheral blood: detection and semi-quantitation by reverse transcription polymerase chain reaction, HEPATOL RES, 14(1), 1999, pp. 1-12
Citations number
22
Categorie Soggetti
Gastroenerology and Hepatology
Journal title
HEPATOLOGY RESEARCH
ISSN journal
13866346 → ACNP
Volume
14
Issue
1
Year of publication
1999
Pages
1 - 12
Database
ISI
SICI code
1386-6346(199903)14:1<1:AOAMIP>2.0.ZU;2-R
Abstract
The presence of a-fetoprotein (AFP) mRNA in mononuclear cells of the periph eral blood was examined in patients with hepatocellular carcinoma (HCC), li ver cirrhosis, chronic hepatitis and normal volunteers using nested reverse transcription polymerase chain reaction (RT-PCR). Then, the AFP mRNA level s were quantitatively analyzed in AFP mRNA-positive patients with HCC. AFP mRNA was detected in 23 of 38 (60.5%) cases with HCC, 2 of 12 (16.7%) cases with cirrhosis, and 1 of 9 (11.1%) cases with chronic hepatitis, respectiv ely. AFP mRNA was not demonstrated in six healthy volunteers. The frequency of positive cases in TNM stage III-IV (16/21, 76.2%) was significantly hig her than that in stage I-II (7/17, 41.2%)(P < 0.05). AFP mRNA was detected more frequently in patients with positive levels of serum protein induced b y vitamin K antagonist II (PIVKA-II) (greater than or equal to 40 mAU/ml, 1 8/25, 72.0%) than in those with negative level (40 mAU/ml, 5/13, 38.5%) (P < 0.05). However, there was no significant correlation between detectable r ate of AFP mRNA and serum AFP level. The frequency of AFP mRNA positive res ult was not closely associated with L. culinaris agglutinin A, a reactive a -fetoprotein (AFP L3). On the other hand, the expression levels of AFP mRNA were measured in 23 AFP mRNA-positive patients with HCC. Quantitative anal ysis of AFP mRNA was performed by comparing the amplification kinetic curve of AFP mRNA with that of beta-actin mRNA as an internal control. The AFP m RNA levels were expressed as Delta cycle values by this method. Delta cycle values were significantly increased in patients with HCC in stages III-IV (4.7 +/- 2.4) compared with patients with HCC in stages I-II (- 1.3 +/- 7.3 ) (P < 0.01). These results suggested that the detection and quantitation o f AFP mRNA in peripheral blood may indicate the hematogenous spreading of t umor cells in patients with HCC. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.