| Objective: Our study object was the patients of HCC, posthepatitis cirrhosis, chronic hepatitis and heathy control, we detected the expression and distribution of HMGB1 in the serum and tissue of patients with HCC.we preliminary discussed the function of HMGB1 in the patients with HCC and analysised its clinical significance, which provided the theoretical basis for the clinical treatment of HCC.Methods: Selecting the case under the definiens and diagnostic standard of HCC, chronic hepatitis B, posthepatitis cirrhosis. The HCC cases fulfilled all above conditions could be brought into the range of study, 40 cases of patients were accorded with those condtions; The posthepatitis cirrhosis cases fulfilled all above conditions could be brought into the range of study, 30 cases of patients were accorded with those condtions; The chronic hepatitis B cases fulfilled all above conditions could be brought into the range of study, 38 cases of patients were accorded with those condtions; The normal control group included 23 cases of heath examination( serum) and 10 cases of hepatic hemangioma(normal liver tissue). 1. Real-Time fluorescence quantitative PCR and ELISA were used to detecte the HMGB1 mRNA and HMGB1 protein expression. then the expression of HMGB1 in the serum at different cliniacal pathology characteristive were analysed.2. Western blot, Real-Time fluorescence quantitative PCR were used to detected the HMGB1 mRNA and HMGB1 protein expression; HMGB1 protein distribution in the tissues of the above-mentioned objects was detected by Immunohistochemistry.Results: 1. ELISA results showed that the serum of HMGB1 protein with HCC, Posthepatitis cirrhosis, chronic hepatitis B and heath examination were 98.85±41.42ng/ml, 66.6±28.99ng/ml, 48.6±18.38ng/ml, 36.6±7.07ng/ml, respectively. The expressions of HMGB1 protein in HCC, Posthepatitis B cirrhosis and chronic hepatitis B were significantly higher than that in heath controls (p<0.05); and the expressions of HMGB1 protein in HCC and Posthepatitis B cirrhosis were significantly higher than that in chronic hepatitis B (p<0.05); the serum of HMGB1 protein in recurrent HCC was 83.85±13.79ng/ml, The expressions of HMGB1 protein in recurrent HCC were significantly higher than that in heath controls (p<0.05).2. Real-Time fluorescence quantitative PCR results showed that the expression levels of HMGB1 mRNA in the serum of HCC were significanctly higher than that in heath examination, and 2-△△CT>2, there was significant different between HCC and heath controls; and the expression levels of HMGB1 mRNA in the serum of Posthepatitis B cirrhosis than that in heath controls, and 2-△△CT>2, there was significant different between Posthepatitis B cirrosis and heath controls.3. There were no relation between HMGB1 protien of the peripheral serum in HCC and the sex, age, degree of differentiation (p>0.05); However, There were closely relation between HMGB1protien of the peripheral serum in HCC and TNM staging, AFP, pathological grade, lymph node metastasis, HBV content, tumor size and hepatitis cirrhosis.4. There were positive correlation between HMGB1 protien and AFP in the peripheral serum of HCC(r=0.823, p<0.05).5. Western blot results showed that the expressions of HMGB1 protein in HCC, Posthepatitis B cirrhosis and chronic hepatitis B were significantly higher than that in normal controls(p<0.05); the expressions of HMGB1 protein in HCC and Posthepatitis B cirrhosis were significantly higher than that in chronic hepatitis B(p<0.05); the expressions of HMGB1 necleus protein in HCC and Posthepatitis B cirrhosis were significantly less than that in normal group(p<0.05); the expressions of HMGB1 cytoplasmic protein in HCC and Posthepatitis B cirrhosis were significantly more than that in normal group (p<0.05); the expressions of HMGB1 protein in recurrent HCC group and recurrent HCC paracancerous group were significantly higher than that in normal controls(p<0.05).6. Real-Time fluorescence quantitative PCR results showed that the expression levels of HMGB1 mRNA in the tissue of HCC were significanctly higher than that in normal liver tissue, and 2-△△CT>2, there was significant different between HCC and normal liver tissue; the expression levels of HMGB1 mRNA in the tissue of Posthepatitis B cirrhosis were significanctly higher than that in normal liver tissue, and 2-△△CT>2, there was significant different between Posthepatitis B cirrhosis and normal liver tissue; the expression levels of HMGB1 mRNA in the tissue of chronic hepatitis B were significanctly higher than that in normal liver tissue, and 2-△△CT>2, there was significant different between chronic hepatitis B and normal liver tissue. Howover, the expression levels of HMGB1 mRNA in the tissue of Posthepatitis B cirrhosis were higher than that in chronic hepatitis B, and 2-△△CT=1.46<2, there was no significant different between Posthepatitis B cirrhosis and chronic hepatitis B; the expression levels of HMGB1 mRNA in the tissue of HCC were higher than that in Posthepatitis B cirrhosis, and 2-△△CT=1.54<2, there was no significant different between HCC and Posthepatitis B cirrhosis; the expression levels of HMGB1 mRNA in the tissue of HCC were higher than that in chronic hepatitis B, and 2-△△CT=2.25>2, there was significant different between HCC and chronic hepatitis B.7.The Immunohistochemistry showed that HMGB1 protein were mainly found in the cytoplasm of HCC, the positive rate of the HMGB1 cytoplasmic protein in HCC was higher than that in normal control. HMGB1 protein were mainly found in the cytoplasm of Posthepatitis B cirrhosis, the positive rate of the HMGB1 cytoplasmic protein in Posthepatitis B cirrhosis was higher than that in normal control, there was significant different between Posthepatitis B cirrhosis and normal control. HMGB1 protein was found both in the nucleus and cytoplasm of chronic hepatitis B, but the mild expression in the nucleus. HMGB1 protein was mainly found in the nucleus of normal liver tissues, all most without expression in the cytoplasm.Conclusions: 1. The contents of HMGB1 protein and the expressions of HMGB1 mRNA in the peripheral blood of HCC patients were significantly higher than that of healthy controls, which indicated that the HMGB1 were closely related to the occurrence and development of HCC.2. The contents of HMGB1 protein and mRNA in the peripheral blood and tissue of Posthepatitis B cirrhosis,chronic hepatitis B and HCC patients were significantly higher than that of healthy controls, which indicated that HMGB1 may be involved in hepatitis cirrhosis and HCC development process of chromic hepatitis B.3. The contents of HMGB1 protein in the cytoplasm and nucleus of chromic hepatitis B patients were more expressed, which indicated that the continued HBV may be stimulate liver cell that synthsised HMGB1 protein, Which promoted hepatocyte proliferation and developed hepatitis cirrhosis.4. The contents of HMGB1 protein in the cytoplasm of HCC and LC patients were more expressed, which indicated that the continued HBV stimulated liver cell synthsised HMGB1 protein that were secreted to cytoplasm or extracellular, thus further demonstrated HMGB1 may be involved in the HCC development process of hepatitis cirrhosis.5. The contents of HMGB1 protein in the peripheral blood and tissue of recurrence HCC patients were significantly higher than that of healthy controls, which indicated that HMGB1 may be involved in the relapse after opteration in the tumor microenviroment.6. The HMGB1protein expression of peripheral blood were closely related to TNM staging, lymph node metastasis, tumor size, which indicated that HMGB1 may be involved in the recurrence,invasion and metastasis of HCC.7. The HMGB1 haved the different localization in the nucleus and cytoplasm, which indicated that the different localization of HMGB1 haved a major impact on its functions.8.The HMGB1 and AFP in the peripheral blood of HCC patients were commonly detected, which helped us to diagnosis, monitor precess and determinate recurrence. |
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