| Objective: Through S-P immuno-histochemical method and Western-Blotting, to detect human aldehyde oxidase(h AOX) in normal liver tissues, cirrhosis, hepatocellular carcinoma(HCC) and pericarcinomatous tissues, and to explore its relationship with liver cancer biological behaviors and clinical significance of prognosis.Methods: HCC tissue samples in 62 cases were taken from pathological specimens resected in liver disease centers of the First Affiliated Hospital of China Medical University and the PAPF General Hospital from August 2003 to May 2008. All patients received no anti-cancer treatment, and all the pathological sections after surgery are based on Ackerman Surgical Pathology(7th edition) diagnostic criteria and given medical results by senior pathologists. 52 cases were males and 10 females, with a median age of 53.7 years(33 to 81 years old); HBs Ag(+) were 47 cases, and HBs Ag(-) 15 cases; AFP≤20ng / ml were 19 cases, and AFP> 20 ng / ml were 43 cases; tumor diameter <3cm were eight cases, 3~5cm in 20 cases, and >5cm were 34 cases; according to histopathological grading standards of hepatocellular carcinoma pathology Volume in Chinese Medical Association clinical Practice Guidelines, 5 cases belonged to grade I, 33 cases grade II, and 24 cases grade III; 27 cases were cirrhosis, and 35 cases without cirrhosis; 17 cases with vascular invasion, and 45 cases without vascular invasion; 48 cases were nodular growth, and 14 cases invasive growth. Another 18 cases of cirrhosis of the liver tissue samples were collected from the liver transplant patients in First Affiliated Hospital of China Medical University, normal liver tissues of 14 cases were taken from accidental death.Immunohistochemical SP method was used to detect 62 cases of hepatocellular carcinoma and pericarcinomatous tissues, 18 cases of liver cirrhosis and 14 cases of normal liver tissues, and at the same time, western blotting was used to detect the expression of h AOX in 3 fresh cases of liver cirrhosis tissues and 12 cases of hepatocellular carcinoma and pericarcinomatous tissues at-70℃ taken from refrigerator, and statistical analysis was conducted through Prism 5.0 software package. h AOX positive expression comparison in liver tissues of each group and the relationship between h AOX and clinical biological behaviors were conducted through chi-square test and Fisher’s exact test, and significant level was 0.05, two-sided test. T test was used to analyze the h AOX expression in the Western blot experiments. Using Kaplan-Meier method and the Log-rank test were used to make survival analysis. P<0.05 was considered statistically significant.Results: Immunohistochemistry results showed that, in 62 cases of hepatocellular carcinoma and pericarcinomatous tissues, 18 cases of liver cirrhosis and 14 cases of normal liver tissues, h AOX expression was the same as that in the cases of normal liver tissues, and the figure was 100%, followed by pericarcinomatous tissues(96.8%; 60/62), and positive rate of hepatocellular carcinoma was the lowest, only 37.1%(23/62). HCC group was Compared with the other groups, and the difference was statistically significant(P <0.05).statistical analysis of correlation between h AOX protein expression levels of HCC specimens and clinicopathological factors of HCC specimens showed that h AOX had different levels of expression in different histological liver specimens, including pathological types: high-expression rate was 80%, middle differentiated and poorly differentiated expression rates were respectively 63.6% and 20.8%, the difference among three groups was statistically significant P<0.05), showing that h AOX was significantly correlated with histopathological grading. In addition, h AOX expression in tumor vascular invasion of HCC was 23.5%, positive rate without vascular invasion was 93.3%, and the difference was statistically significant. At the same time, the existence of different tumor growth pattern resulted in different h AOX expression, h AOX expression in nodular HCC growth was 93.8%, and positive expression in HCC invasive growth was 28.6%, showing the difference was statistically significance(P <0.05). In addition, h AOX positive expression in males and females was respectively 51.9% and 60.1%, showing the difference was not statistically significant(P >0.05); the age of 50 was regarded as the dividing line, the h AOX positive expression rate in patients under 50 was 51.4%, and the expression in patients over age 50 was 53.0%, showing that the difference was not statistically significant(P >0.05). h AOX expression was not significantly correlated with tumor size, AFP level and HBs Ag- / + and presence of cirrhosis(P >0.05).5-year survival rate of the patients with h AOX(+) expression was significantly higher than h AOX(-) expression patients(X2 = 5.829, P <0.05). Through the survival analysis by Kaplan-Meier method, the results suggested that h AOX(+) expression patients had significantly better prognosis than h AOX(-) expression cases(Log-rank test, P <0.05).The study on 3 liver cirrhosis tissues and 12 pairs of hepatocellular carcinoma and pericarcinomatous tissues showed that h AOX expression in hepatocellular carcinoma and liver cirrhosis tissues was significantly higher than that in liver cirrhosis tissues. As to h AOX expression in hepatocellular carcinoma tissues, seven cases had no expression(56.3%), three cases had low expression(31.2%), and only two cases(12.5%) was not statistically significant compared with that in pericarcinomatous tissues, and three cases of liver cirrhosis tissues showed high expression(100%).Conclusion: h AOX expression in normal liver, cirrhosis, and pericarcinomatous liver tissues was higher than that in HCC tissues, which might be connected with vascular invasion and tumor growth pattern of liver cancer. h AOX is likely to become the major targeted drug treatment of liver cancer in the future. |
