Quantitative Analysis Of AFP MRNA Of The Human Hepatocellular Carcinoma By FQ-PCR And Its Significance

Hepatocellular carcinoma(HCC) is one of the most rapidly fatal neoplasms worldwide. The only hope of long-term survival for HCC patients is tumor resection or liver transplantation. However, these therapeutic options are severely limited by the frequent presence of recurrence/metastasis. Over half of those who undergo apparently successful resection subsequently develop recurrence, which may presumably arise from tumor cells that are shed during or after surgery. Early diagnosis of recurrence/metastasis may permit the selection of patients for surgical resection. This could also be valuable for monitoring patients' response to different therapies.Human AFP gene,consisting of 14 exons tandemly located at the fourth chromosome, are 17kb long. AFP, an oncofetal protein, is synthesized mainly in hepatocytes of fetal liver and yolk sac, and AFP production by hepatocytes is rapidly reduced from the time of birth in inverse proportion to an increased synthesis of albumin. AFP mRNA has lately been detected in tumor tissue of HCC but not in nontumorous tissue, and its gene expression is reported to be modulated by hypomethylation of the AFP gene. Hitherto, increased serum AFP concentration, a tumor marked in patients with HCC ,is used in screening for HCC, but it cannot predict metastasis of HCC.The presence of circulating HCC cells, which may be released from tumor foci into the circulation,could be an indicator of hematogenous spread of tumor cells leading to extrahepatic metastasis of HCC.In this study, in an attempt to demonstrate the Significance of AFP mRNA ,we try to analyze it by fluorescence quantitative ( FQ ) - polymerase chain reaction (PCR) combined with nest PCR.Part 1: Detection of AFP mRNA in Peripheral Blood of Patients withHepatocellular Carcinoma by FQ-PCR and its Significance1. Objective:To evaluate AFP mRNA expression in peripheral blood of the patients with hepatocellular carcinoma(HCC)and its significance.2. Materials and Methods2.1 Patientsthe samples were obtained from 51 patients with hepatocellular carcinoma, 10 metastatic cancer of liver, 18 non-HCC malignant tumors, 16 chronic liver diseases in the Second Affiliated Hospital of Zhejiang University Medical school between 2001 and 2002.HCC patients were regularly assessed for clinical stage and metastasis or recurrence by histopathological features and measurement of serum AFP level,ultrasound examination with computed tomography, 43 case were male and 7 were female.the mean age was 41 years. 39 cases underwent operation.2.2 Methodsfluorescence quantitative ( FQ ) - polymerase chain reaction (PCR) combined with nest PCR3. Results:AFP-mRNA was not detected in the patient with non-HCC malignant tumors but in 2 of 10 metastatic cancer of liver (20.0%). The abnormal expression of AFP-mRNA in the patients with HCC (37.3% 19/51) was higher than those with chronic liver diseases ( 6.3 % 1/16, P<0.05 ) , and the detective rate of AFP-mRNA in the blood seemed to be correlated with the clinical stages of HCC, the presence of extrahepatic metastasis, portal vein thrombosis but not with serum AFP level andtumor size.4. Conclusions;The abnormal expression of AFP-mRNA is detected in peripheral blood by FQ-PCR , which might predicate hematogenous spreading metastasis of tumor cells in the patients with HCC, It may be of clinical significance for the diagnosis,staging, detection of extrahepatic metastasis and prognosis of HCC patients. Keywords: FQ-PCR; Liver neoplasms ; AFP-mRNAPart2: Quantitative analysis of AFP mRNA of the human hepatocellularcarcinoma(HCC)tissue,per-HCC tissues and Peripheral Blood by FQ-PCR and its Significance1. Objective:To evaluate AFP-mRNA expression in the human hepatocellular carcinoma (HCC) tissue, per-HCC tissues and Peripheral Blood and its significance.2. Materials and MethodsAFP mRNA was detected by FQ-PCR in the human hepatocellular carcinoma(HCC)tissue,per-HCC tissues and Peripheral Blood, the samples were obtained from 30 patients w...