1. INTRODUCTION & OBJECTIVES Introduction: The carcinogenesis of hepatocellular carcinoma (HCC) is a multifactorial, multistep and complex process. Its prognosis is poor and early diagnosis and monitoring of metastasis of HCC is of the utmost importance. Circulating AlphaFetoProtien (AFP) mRNA has been proposed as a marker of HCC cells disseminated into the circulation. The specificity of this molecular marker and its correlation with the main HCC clinico-pathological parameters remains controversial. In recent year; several different multimarkers assays have been developed for the detection of hepatoma cells in the peripheral blood of patients with HCC. Objectives: In this study; we examined the expression of combined multimarker assay of cancer specific markers Melanoma Antigen Gene MAGE 1 and MAGE 3 mRNAs and liver specific marker AFP mRNA in blood specimens obtained from patients with primary HCC and also from non HCC patients and control group by nested reverse transcriptase polymerase chain reaction (RT-PCR) to offer simple method with high specificity and sensitivity for detection of the circulating hepatoma cells. Subjects: In this study 58 patients and 15 matched control subjects; the patients divided into 4 groups; group A; patients with primary HCC diagnosed by histology, imaging, serum AFP,.. (n =32). Group B; patients with cirrhosis with no evidence of HCC (n =12). Group C; patients with metastatic cancer in liver (n =14). Group D; 15 healthy volunteer with normal liver function with no history or evidence of diseases. The staging of HCC was carried out according to the (Tumor / Node / Metastasis ) TNM classification. Methods: Peripheral blood samples were obtained from all subjects; MAGE-1 and 3 mRNAs and AFP mRNA were detected by nested RT-PCR. Our results indicate that a multimarker nested RT-PCR assay with cancer-specific markers such as MAGE-1 and MAGE-3 in combination with a hepatocyte-specific AFP marker may be a promising diagnostic tool for monitoring hepatocellular carcinoma patients. Since nested PCR utilizes a couple of internal primers to reamplify the specific PCR product, it exhibit higher sensitivity, stronger specificity and lower false positive occurrence as compared to single RT. References RESULTS CONCLUSIONS The positive rates of MAGE-1 and MAGE- 3 mRNAs and AFPmRNA were 18/32 (56.3%) and 15/32 (46.9%); 19/32 (59.4%) respectively in the primary HCC patients. 4/12 (33.3%) patients with cirrhosis positive for AFPmRNA and 5/14 (35.7%); 4/14 (28.6%); with metastatic cancer in liver positive for MAGE- 1 and 3 mRNAs. The positive rate of MAGE-1 and 3 mRNAs was correlated with TNM clinical stages; tumor size and extrahepatic metastasis (P<0.05). The positive rate of AFPmRNA was correlated with TNM clinical stages but no correlation was found between tumor size or tumor number. No samples of clinical controls show detectable levels of MAGE-1 and MAGE- 3 mRNAs and AFPmRNA in their peripheral blood.tumor SUBJECTS & METHODS Multiple Molecular Markers Specific Circulating Hepatoma Cells: Enhanced Detection of HCC Salwa H. Teama1; Sara H. A. Agwa 2; Noha A. ELNakeeb3 Medical Research Center; Molecular Biology Unit; Ain Shams University, Cairo, Egypt1&2 Department of Internal Medicine; Ain Shams University, Cairo, Egypt 3 1- Marrero JA, Pelletier S. Hepatocellular carcinom (2006). Clin Liver Dis; 10 (2): 339- 51. 2- Liu Yu-Hui, Zhou Rou-Li and Rui Jing (1998). Detection of hepatoma cells in peripheral blood of HCC patients by nested RT-PCR; 4(2): 103-108. 3-Yao D Dong Z et al., (2007). Specific molecular marker in HCC. Hepatobil.Panc.Dis.Int; 6(3):241-247. 4- Abd El- Haamid NM (2008). Priority consideration in early laboratory diagnosis of HCC. IJID; Vol3: 3199. Fig. 1: Expression of MAGE-1; MAGE-3 mRNAs and AFPmRNA in different group Available from;http://www.ejmhg.com/article/S1110-8630(12)00087-0/abstract