Biological Etiology Of Primary Liver Cancer

PartⅠAssociation between the Presence of HBV, HCV Infectionin HCC and Clinic, HistologyObjective To study the biological behaviour of HBV and HCV infection in HCC.Methods The patterns of HBsAg and HCV in 100 cases of hepatocellularcarcinoma(HCC) and their surrounding liver tissues were studied on paraffin-embededsections with immunohistochemistry technique, pericarcinomatous tissueswere staged; AFP, Hyaluronic acid (HA), procollagenⅢ(PⅢP), typeⅣcollagen(C-Ⅳ), laminin(LN) were detected by radioimmunoassay; the activities of Tlymphocytesubset and NK cell were examined; the correlation were studied.Results The levels of AFP, HA, C-Ⅳ, LN in HBV and HCV coinfection is the highestin the four groups. The levels of AFP, HA, C-Ⅳ, LN in the group which is notinfected by HBV, HCV is the lowest in the four groups. HBV, HCV expression werepositively correlated with AFP, HA, LN and C-Ⅳ. The immunologic derangement inHCC were concemed with HBV, HCV infection.Conclusions The background of HBV, HCV virus infection had a direct influence onthe levels of AFP, HA, C-Ⅳ, LN, the fibrotic staging. On the one hand, virus infectionis one of the reasons which cause the liver cancer; on the other hand, perennialvirusemia will aggravate pathological changes of liver tissue. The antivirusintervening treatment of hepatitis is significant for the prognosis of liver cancer.PartⅡHelicobacter Infection in Hepatocellular Carcinoma TissueObjective Histology with standard and culture, electron microscope, molecularbiology were used to detect the presence of Helicobacter in hepatocelluer carcinomatissue, to look for the disease mechanism, prevention and cures of the experiment and theorieses.Methods 1,Liver samples from thirty-four patients with hepatocelluer carcinoma(HCC) diagnosed by histopathology were studied. Twenty-two patients with otherliver diseases(five with liver trauma, seven with cavernous liver hemangioma, sixwith liver cyst and four with hepatolithiasis), Twenty-five patients with gastriccancer, fifteen with colonic cancer and fifteen with myoma of uterus served ascontrols. Helicobacter spp. in biopsy from the studied subjects were detected bypolymerase chain reaction (PCR) with Helicobacter spp. 16S rRNA primers.Amplified products were identified by Southern hybridization and were sequencedfurther. Besides, the other genes(vacA,cagA) that were specific for Helicobacterpylori (H. pylori) was also detected by PCR. 2,The samples was cranked outpathologic slice for in situ hybridization of Helicobacter. 3,Helicobacter spp. in thebiopsies was observed by scanning electron microscope directly.4,The samples werecultured on the Columbia agar plates with microaerobic techniques. Helicobactercolonies were identified further with the methods, which included their typicalmorphology, characteristic appearance on Gram staining, positive urease test,observed by transmission electron microscope directly. Another way to test theHelicobacter colonies was extracting DNA to examine Helicobacter spp. 16S rRNA,cagA and vacA genes by PCR, then the PCR products were sequenced.Results 1.PCR showed that the positive rate of HCC and gastric cancer samples was64.71% and 72%, respectively, while the controls were negative(P＜0.01). 2.ThePCR-amplified products were identified by Southern hybridization and sequenced.The homology to 16S rRNA of H. pylori, was 97.80%. 3.Only three HCC samples andtwo gastric cancer samples of the cagA genes were detected. None of the samplesreacted with primers for vacA in the two groups. 4.The positive samples were verifiedby in situ hybridization for Helicobacter spp. 16S rRNA-mRNA, and proved to be H.pylori, which include twenty-two HCC samples, seventeen gastric cancer samples andone colonic cancer sample. There was no statistical significance between HCC andgastric cancer (P＞0.05), but the positive rate of HCC and the other controls hadstatistical significance (P＜0.01). 5.The cocci was observed in adjacent hepatocytes ofthe two HCC samples by SEM. The bacterium was also found in four gastric cancersamples. 6.The positive rates of Helicobacter cultured in HCC and gastric cancer were 8.82%(3/34) and 24%(6/25), respectively. Helicobacter microorganisms wereidentified further by typical appearance on Gram staining, positive urease test andcharacteristic colony morphology on TEM.Conclusion 1,Helicobacter infection exists in liver tissues of HCC patients and theinfectious rate was high. 2,There are an association between Helicobacter infectionand HCC...