| Objective: Thy-1, CD34, CK-7, AFP as a marker of hepatocarcinoma stem cells has been preliminarily confirmed, while the E-cadherin, VEGF is the reaction has been recognised an tumor proliferation, mesastasis, and used to evaluate the prognosis of the portein. In this study, The immunohistochemical staining was used to study the relationship between the expression of the stell cell marker and the mesastasis of markers in different tissues of the liver. Through analysis cancer stem cell marker and the transfer of indicators to further explore the correlation between the cancer stem cells and metastasis and prognosis.Methods:1 Samples collection: 50 specimens of HCC, 50 specimens of adjacent non-cancerous cirrhosis hepatic tissue and 20 specimens of normal hepatic tissue were obtained from the fourth hospital of Hebei Medical University without clinical treatment before operation.2 Imunostaining: The samples were fixed by 10% formaldehyde, embeded in paraffin after excision. Thy-1, CD34, CK-7, AFP and VEGF E-Cadherin were stained immunohistochemistry, and two pathologist without backgroud on these samples graded these oncogenes expression and proliferation on each tissue.3 Follow up: The shoerest time followed up was one month and the longest time was twenty two month.Resluts:1 Survival analysis of the clinical and pathological data in postoperative HCC patients: Age, AFP, the size of tumor, HbsAg and HbeAb were no statistical difference in postoperative HCC patients. The clinical stage (χ2=4.103, P=0.043), the pathological classification (χ2=9.435, P=0.009), AFP (χ2=15.802, P=0.000) and the portal vein tumor thrombosis(χ2=5.148, P=0.023)were the significant prognostic factors in postoperative HCC patients.2 The expression of Thy-1was in cytoplasm and cellular membrane. The expression of Thy-1 had no significant difference refered to sex, AFP, HBeAb, HBsAg and the integrity of the tumor's envelope based on statistical analysis. The expression of Thy-1 in HCC was significantly higher than that in adjacent non-cancerous hepatic tissue and in normal hepatic tissue (Z=-7.018 and Z=-6234, P=0.000), and no statistical difference existed between in adjacent non-cancerous hepatic tissue and in normal hepatic tissue. The expression of Thy-1 was positively correlated with hepatic carcinogenesis. The expression of Thy-1 in later clinical stage in HCC was stronger than that in early clinical stage (χ2=10.771, P=0.029; r=0.394, P=0.005). The expression of Thy-1 in the in poorly differentiated tissue was stronger than that well-differentiated HCC about the pathological classification (χ2=9.357, P=0.009; r=0.409, P=0.003). The expression of Thy-1 in the tissue with the portal vein tumor thrombosis was stronger than that without the portal vein tumor thrombosis (Z=-3.041, P=0.002; r=-0.434, P=0.001). This experiment demonstrated that The expression of Thy-1 was positively correlated with the VEGF expression in the hepatic carcinogenesis (r=0.516, P=0.000) and with the negatively correlated with the E-cadherin expression in the hepatic carcinogenesis (r=-0.161, P=0.030). The patients in whom Thy-1 was negatively and weakly expressed had 17.85 months'mean survival time, and the cumulative survival rate was 73.3%. The patients in whom Thy-1 moderately and strongly expression had 9.16 months'mean survival time, and the cumulative survival rate was 9.1%. Thy-1 expression was stronger, the survival rate was lower, and the prognosis was worse (By single variable Log rank test,χ2=14.213, P=0.000).3 The expression of CK7 was in cytoplasm and cellular membrane. The expression of CK7 had no significant difference refered to sex, AFP, HBeAb, HBsAg and the integrity of the tumor's envelope based on statistical analysis. The expression of CK7 in HCC was significantly higher than that in adjacent non-cancerous hepatic tissue and in normal hepatic tissue (Z=-4.897 and Z=-514, P=0.000), and no statistical difference existed between in adjacent non-cancerous hepatic tissue and in normal hepatic tissue. The expression of CK7 was positively correlated with hepatic carcinogenesis. The expression of CK7 in later clinical stage in HCC was stronger than that in early clinical stage (χ2=18.060, P=0.001; r=-0.581, P=0.000). CK7 expression in the poorly-differentiated HCC about the pathological classification was stronger than that in well differentiated tissue (χ2=11.279, P=0.004; r=0.470, P=0.001). The expression of CK7 in the tissue with the portal vein tumor thrombosis was stronger than that without the portal vein tumor thrombosis (Z=-2.654, P=0.008; r=0.379, P=0.007). This experiment demonstrated that The expression of CK7 was positively correlated with the VEGF expression in the hepatic carcinogenesis(r=0.494, P=0.000) and with the negatively correlated withtheE-cadherin expressed in the hepatic carcinogenesis(r=-0.374, P=0.002). The patients in whom CK7 negatively and weakly expression had 15. 70 months'mean survival time, and the cumulative survival rate was 58.8%. The patients in whom CK7 was moderately and strongly expressed had 10.01months'mean survival time, and the cumulative survival rate was 15.0%. CK7 expression was stronger, the survival rate was lower, and the prognosis was worse (By single variable Log rank test,χ2=6.272, P=0.012).4 The expression of AFP was in cytoplasm and cellular membrane. The expression of AFP had no significant difference refered to sex, AFP, HBeAb, HBsAg and the integrity of the tumor's envelope based on statistical analysis. The expression of AFP frequency in HCC was significantly higher than that in adjacent non-cancerous hepatic tissue and in normal hepatic tissue (Z=-2.790 and Z=-4.881, P=0.000), and no statistical difference existed bwteen in adjacent non-cancerous hepatic tissue and in normal hepatic tissue. The expression of AFP was positively correlated with hepatic carcinogenesis. The expression of AFP in later clinical stage in HCC was stronger than that in early clinical stage (χ2=10.917, P=0.028; r=0.359, P=0.011). The expression of AFP in the poorly-differentiated HCC about the pathological classification was stronger than that in well differentiated tissue (χ2=14.462, P=0.003. r=0.461, P=0.000). The expression of AFP in the tissue with the portal vein tumor thrombosis was stronger than that without the portal vein tumor thrombosis (Z=-4.216, P=0.000; r=-0.602, P=0.000). The expression of AFP was positively correlated with the VEGF expression in the hepatic carcinogenesis(r=0.419, P=0.003) and with the negatively correlated with the E-cadherin expressed in the hepatic carcinogenesis(r=-0.332, P=0.005) The patients in whom AFP was negatively and weakly expressed had 18.73 months'mean survival time, and the cumulative survival rate was 78.6%. The patients in whom AFP was moderately and strongly expressed had 9.21months'mean survival time, and the cumulative survival rate was 8.7%. AFP expression was stronger, the survival rate was lower, and the prognosis was worse (By single variable Log rank test,χ2=15.772, P=0.000).5 The expression of CD34 was in cytoplasm and cellular membrane. The expression of CD34 has no significant difference refered to clinical stage, sex, age, the size of tumor, AFP, HBeAb, HbsAg, the portal vein tumor thrombosis and the integrity of the tumor's envelope based on statistical analysis. The expression of CD34 in HCC was significantly higher than that in adjacent non-cancerous hepatic tissue and in normal hepatic tissue (Z=-3.708, P=0.000 and Z=-2.913, P=0.003), and no statistical difference existed bwteen in adjacent non-cancerous hepatic tissue and in normal hepatic tissue. The expression of CD34 was positively correlated with hepatic carcinogenesis. CD34 expression in later clinical stage in HCC was stronger than that in early clinical stage (χ2=12.005, P=0.017. r=0.452, P=0.012). The expression of CD34 in the poorly-differentiated HCC about the pathological classification was stronger than that in well differentiated tissue (χ2=8.487, P=0.014; r=0.286, P=0.042). The expression of CD34 in the tissue with the portal vein tumor thrombosis was stronger than that without the portal vein tumor thrombosis (Z=-2.633, P=0.008; r=0.376, P=0.007). The expression of CD34 positively correlated with the VEGF expression in the hepatic carcinogenesis (r=0.356, P=0.007) and with the negatively correlated with the E-cadherin expressed in the hepatic carcinogenesis (r=-0.541, P=0.000). The patients in whom CD34 was negatively and weakly expressed had 17.16 months'mean survival time, and the cumulative survival rate was 63.2%. The patients in whom CD34 was moderately and strongly expressed had 8.42 months'mean survival time, and the cumulative survival rate was 11.8%. CD34 expression was stronger, the survival rate was lower, and the prognosis was worse (By single variable Log rank test,χ2=14.17, P=0.000).6 The expression of VEGF was in cytoplasm and cellular membrane. The expression of VEGF had no significant difference refered to clinical stage, sex, age, the size of tumor, AFP, HBeAb, HbsAg, the portal vein tumor thrombosis and the integrity of the tumor's envelope based on statistical analysis. The expression of VEGF in HCC was significantly higher than that in adjacent non-cancerous hepatic tissue and in normal hepatic tissue (Z=-5.494, P=0.000 and Z=-3.959, P=0.000), and no statistical difference existed between in adjacent non-cancerous hepatic tissue and in normal hepatic tissue. The expression of VEGF was positively correlated with hepatic carcinogenesis. VEGF expression in later clinical stage in HCC was stronger than that in early clinical stage ((χ2=11.880, P=0.018; r=0.383, P=0.008).VEGF expression in the poorly-differentiated HCC about the pathological classification was stronger than that in well differentiated tissue (χ2=10.196, P=0.006, r=0.455, P=0.001). The expression of VEGF in the tissue with the portal vein tumor thrombosis was stronger than that without the portal vein tumor thrombosis (Z=-3.166, P=0.002; r=0.452, P=0.001). The patients in whom VEGF was negatively and weakly expressed had 16.05 months'mean survival time, and the cumulative survival rate was 57.9 %. The patients in whom VEGF was moderately and strongly expressed had 6.27 months'mean survival time, and the cumulative survival rate was 11.0%. VEGF expression was stronger, the survival rate was lower, and the prognosis was worse (By single variable Log rank test,χ2=20.310, P=0.000).7 The expression of E-cadherin in cytoplasm and a few in cellular nucleus.The expression of E-cadherin had no difference refered to clinical stage, sex, age, the size of tumor, AFP, HBeAb, HbsAg, the portal vein tumor thrombosis and the integrity of the tumor's envelope based on statistical analysis. The expression of E-cadherin in HCC was significantly weaker than that in adjacent non-cancerous hepatic tissue and in normal hepatic tissue (Z=-5.210, P=0.000; and Z=-4.832, P=0.000), and no statistical difference existed bwteen in adjacent non-cancerous hepatic tissue and in normal hepatic tissue. The expression of E-cadherin was negatively correlated with hepatic carcinogenesis. The expression of E-cadherin in early clinical stage in HCC was stronger than that in later clinical stage ((χ2=12.318, P=0.015; r=-0.471, P=0.001). The expression of E-cadherin in the well-differentiated HCC about the pathological classification was stronger than that in poorly differentiated tissue (χ2=16.361, P=0.000; r=-0.551, P=0.000). The patients in whom E-cadherin negatively and weakly expressed had 20.14 months'mean survival time, and the cumulative survival rate was85.7%. The patients in whom E-cadherin moderately and strongly expressed had 10.56 months'mean survival time, and the cumulative survival rate was 23.3%. E-cadherin expression was stronger, the survival rate was higher, and the prognosis was better (By single variable Log rank test,χ2=7.964, P=0.005).Conclusions:1 The clinical stage, the pathological classification and the portal vein tumor thrombosis were the significant prognostic factors in postoperative HCC patients. In the patients in later clinical stage with the portal vein tumor thrombosis whose hepatic tissue was bad-differentiated, the survival rate was lower, the prognosis was worse.2 The expression of Thy-1, CK-7, CD34, AFP were positively correlated with hepatic carcinogenesis. The expression of Thy-1, CK-7, CD34, AFP were stronger in the late clinical stage, poorly differentiated tissueand with the portal vein tumor thrombosis. While the survival rate was shorter, and the prognosis was poorer.3 The expression of the VEGF was positively correlated with hepatic carcinogenesis.The expression of the VEGF was stronger in bad-differentiated tissue. While the survival rate was lower, and the prognosis was poorer. Conversely, The expression of the E-cadherin was stronger While the survival rate was lower, and the prognosis was poorer.4 There were positively correlations between the expression of Thy-1, CK-7, CD34, AFP and VEGF.while nagertively correlations between the expression of Thy-1, CK-7, CD34, AFP and VEGF and E-cadherin expression. It suggested that liver cancer stem cells play an important role in liver cancer occurrence and development, especially liver cancer recurrence and metastasis. It is expected to become the treatment and assessment of prognosis of liver cancer targets.
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