Detecting AFP mRNA in peripheral blood of the patients with hepatocellular carcinoma, liver cirrhosis and hepatitis.
BACKGROUND
The low frequency of disseminated carcinoma cells in the blood now makes immunomagnetic bead sorting and reverse transcriptase-polymerase chain reaction (RT-PCR) technique more popular.


METHODS
Three milliliters of peripheral blood were collected from 91 patients and 18 normal donors. The circulating carcinoma cells were enriched with CD45 and Ber-EP4 immunomagnetic beads. The alpha-fetoprotein (AFP) mRNA was amplified with nested RT-PCR.


RESULTS
The total positive detection rate was 72.1%, 43.8%, 25.0%, 100%, and 66.7% in patients with hepatocellular carcinoma (HCC) untreated, liver cirrhosis (LC), hepatitis, metastasis liver cancer, and postsurgery of hepatocellular carcinoma, respectively. There was a significant difference among the patients with HCC, LC and hepatitis (HCC vs. LC, P<0.05; HCC vs. hepatitis, P<0.01) and between Class A and B of the HCC patients (P<0.05). Meanwhile, AFP mRNA was markedly expressed in HCC patients compared to the patients with no HCC (LC and hepatitis). The levels of aspartate transaminase (AST) and gamma-glutamyltranspeptidase (GGT) were significantly different in AFP mRNA-positive patients with autoimmune chronic active hepatitis B (CAHB) or LC in contrast to the corresponding negative patients.


CONCLUSIONS
Combining negative and positive immunomagnetic bead sorting and RT-PCR technique can effectively detect circulating tumor cells. AFP mRNA is a more reliable marker of metastasis compared to serum AFP.
Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai, China.
Feng
F
Yao
Jun-Ming
JM
Guo
Chang-Feng
CF
Xu
Yan-Lu
YL
Lou
Bing-Xiu
BX
Xiao
Wen-Hong
WH
Zhou
Jian
J
Chen
Yao-Ren
YR
Hu
Zhong
Z
Liu
Guo-Fan
GF
Hong
D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000032:analysis; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D005260:Female; D006505:Hepatitis / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D006801:Humans; D008103:Liver Cirrhosis / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009367:Neoplasm Staging; D012333:RNA, Messenger / Q000097:blood* / Q000235:genetics; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000235:genetics*
D015415:Biomarkers; D012333:RNA, Messenger; D000509:alpha-Fetoproteins