Anticorpi anti nucleari e cellule tumorali circolanti nello studio dell'epatocarcinoma

Background: Hepatocellular carcinoma (HCC) is the most frequent primary liver cancer and occurs, in most cases, in patients with chronic liver disease (70-90% of HCC cases), presenting itself in a form multifocal at diagnosis in 75% of cases. The incidence of primary liver cancer is related primarily to the spread of hepatitis B and C, which has increased between 1950 and 1980 in developed countries. HCC is the most common type of liver cancer worldwide, with a prevalence below that of hepatitis B and C. The incidence is higher in Asia and in sub-Saharan countries, while it is relatively less common in Europe and North America. At European countries where there are the most cases, they are those of the Mediterranean, including Italy. The onset of HCC is usually detected at a distance of 20 to 30 years from the beginning of liver damage and the average age of onset is 60-65 years. Patients with cirrhosis have a higher risk of developing HCC: 90-95% of patients with HCC have liver disease or cirrhosis of the base, but are also recorded cases of non-cirrhotic liver of HCC. The risk factors for the onset of hepatic cancer are to be sought in pre-existing diseases of the liver such as: cirrhosis, chronic infection with hepatitis B virus (HBV) and hepatitis C (HCV), alcohol abuse and cigarette smoking, obesity (fatty liver) and aflatoxins. Several works in the literature reveal that antinuclear antibodies (ANA), first level test used in the diagnosis of systemic autoimmune disorders (MAIS), may be positive in patients with cancer but, do not manifest symptoms that match the MAIS. In the peripheral blood of some patients with cancer is possible found Circulating Tumor Cells (CTCs), particularly those with metastatic disease. The clinical impact of this phenomenon has long been debated. CTCs have been measured in patients with cancer to analyse metastatic mechanisms or in the hope of developing clinical applications for diagnosis and therapy.Aim: The purpose of this study are essentially two: 1) to detect anti – nuclear antibodies atypical (not commonly associated with the MAIS) as possible tumor markers being HCC . 2) the HCC having a malignant tumor metastases can give. For this reason it was decided to look for the presence of circulating tumor cells in patients with hepatocellular carcinoma , so as to highlight possible parameters that give more information on the state of the disease and especially on its evolution.Methods: In this work we were enrolled 75 patients diagnosed with hepatocellular carcinoma, some of them were hospitalized at the departments of Internal Medicine and Pathology, University of Sassari others were outpatients , and compared with 19 healthy controls. These patients have been tested in the laboratory for research of the bands " atypical”. Another 15 patients with HCC before diagnosis of non metastatic and untreated were included in the study for the detection of circulating tumor cells and compared with 12 healthy controls. The antibodies anti nucleus of 75 patients with HCC was performed by indirect immunofluorescence (IFI). Of these 75 patients 65 were tested for the detection of protein bands "atypical" by Western Blot (WB). The detection of circulating tumor cells has instead been performed by flow cytometry.Results: The results show positive for ANA detection of 52% with a prevalence of males than females. The panel fluoroscopic most represented is homogeneous followed by granular and nucleolar. The analysis of the WB show that 63% of the patients tested positive for this test it is 63.3% and show the presence of protein bands "atypical". Those are statistically significant: 104 kDa, 54.7 kDa, 37.7 kDa and 53 kDa. “Atipycal” band of 53 kDa was not taken into account because was present in healthy controls. The flow cytometry put in evidence the presence of a high number of CTC in the 15 patients with non-metastatic HCC and not treated with any type of therapy.Conclusions: the detection of ANA in patients with hepatocellular carcinoma showed no additional information that was not already present in literature. These ones fact confirms that the ANA are present in this tumor especially in males. Unlike the WB analysis they were identified 3 "atypical" band protein present in patients with hepatocellular carcinoma: 104 kDa, 37.7 kDa and 54.7 kDa. These bands have not been described in terms of research literature yet. For the research of CTC has been possible to distinguish them from other cell populations using specific markers labeled with different fluorophore: CD 45, CD 34, CD 105 and EpCAM. In patients studied was found a high concentration of CTC. Inside the group of the patients positive for the presence of CTCs, some patients show a higher number of these. On the basis of clinical information it is not possible yet describe why some of them have a higher number of CTC.