Quantitative analysis of aldolase A mRNA in liver discriminates between hepatocellular carcinoma and cirrhosis.
BACKGROUND
Chronic liver diseases can progress to cirrhosis and to hepatocellular carcinoma. Timely and unequivocal recognition of the neoplastic evolution of cirrhosis is critical. To this aim, we used a noncompetitive reverse transcription-PCR procedure to analyze aldolase A mRNA in liver tissue from patients with chronic liver diseases at different stages.


METHODS
We studied 12 patients with hepatocellular carcinoma, 19 patients affected by chronic hepatitis C or cirrhosis, and 7 healthy controls. Aldolase A mRNA was reverse-transcribed to cDNA, which was then amplified by PCR. The amplified segments were "read" with a novel dot-blot procedure. A calibrator with the same sequence, synthesized in vitro using a T7 phage promoter, was processed at scalar dilutions in parallel to the target samples to generate a calibration curve and so quantify the target mRNA (detection limit, 0.03 amol; linearity spanning five orders of magnitude).


RESULTS
Aldolase A mRNA was approximately 10-fold higher in liver biopsies from patients with hepatocellular carcinoma vs patients with chronic hepatitis C or cirrhosis, and healthy individuals. Furthermore, aldolase A mRNA concentrations were 1.2- to 21.3-fold higher in 12 liver biopsies compared with the paired surrounding cirrhotic tissue.


CONCLUSIONS
The quantitative analysis of liver tissue aldolase A mRNA differentiates between nonneoplastic chronic liver diseases and hepatocellular carcinoma, which suggests that it has diagnostic potential.
Castaldo
G
G
Dipartimento di Biochimica e Biotecnologie Mediche and CEINGE scarl, Cattedra di Gastroenterologia, Facoltà di Medicina e Chirurgia, Università di Napoli "Federico II", I-80131 Naples, Italy.
Calcagno
G
G
Sibillo
R
R
Cuomo
R
R
Nardone
G
G
Castellano
L
L
Del Vecchio Blanco
C
C
Budillon
G
G
Salvatore
F
F
D006528:Carcinoma, Hepatocellular / Q000175:diagnosis*; D003937:Diagnosis, Differential; D005260:Female; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis* / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000032:analysis / Q000235:genetics; D008099:Liver / Q000737:chemistry*; D008103:Liver Cirrhosis / Q000175:diagnosis*; D008113:Liver Neoplasms / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000032:analysis*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
D007527:Isoenzymes; D012333:RNA, Messenger; D005634:Fructose-Bisphosphate Aldolase