| Objective: To detect the expression of KiSS-1, KiSS-1R and MMP-9 in hepatocellular carcinoma(HCC) and study the correlation of KiSS-1, KiSS-1R and MMP-9 expression to invasion and metastasis of HCC.Methods: The expression of KiSS-1, KiSS-1R mRNA in 33 cases of HCC samples, 2 portal vein tumor embolus samples, 26 non-neoplastic adjacent liver tissue samples and 13 non-neoplastic distant liver tissue samples were detected by RT-PCR, and the correlations of KiSS-1, KiSS-1R mRNA to clinicopathologic parameters of HCC were analyzed. The processing of tissue chip would be modified. Tissue chips were constructed, which contained HCC, non-neoplastic adjacent liver tissues, non-neoplastic distant liver tissues, normal liver tissues and intrahepatic metastasis lesions. The expression of KiSS-1, MMP-9 protein were determined by tissue chips, immunohistochemistry(IHC) and semi-quantitative image analysis in 150 HCC samples, 137 non-neoplastic adjacent liver tissue samples, 98 non-neoplastic distant liver tissue samples, 16 normal liver tissue samples and 37 intrahepatic metastasis lesion samples . The expression of KiSS-1, MMP-9 mRNA were evaluated by tissue chips , in situ hybridization (ISH) and semi-quantitative image analysis in 137 HCC samples, 119 non-neoplastic adjacent liver tissue samples, 85 non-neoplastic distant liver tissue samples, 16 normal liver samples and 28 intrahepatic metastasis lesion samples. The correlations of KiSS-1, MMP-9 to clinicopathologic parameters of HCC were also analyzed.Results:1,The results of RT-PCR showed that compared with the non-neoplastic adjacent liver tissues,non-neoplastic distant liver tissues and normal liver tissues, the expression of KiSS-1 mRNA in HCC was significantly lower(P<0.01), which decreased with the advance of the pathologic grade and clinical stage in HCC. The expression of KiSS-1 mRNA in HCC with metastasis was lower than that in those without metastasis(P<0.05). KiSS-1 mRNA was not detected in 2 portal vein tumor embolus samples, but it was not associated with sex, age, AFP lever, tumor size, capsule invasion and histological type(P>0.05). The expression of KiSS-1R mRNA was not associated with all clinicopathologic parameters of HCC(P>0.05), however a negative correlation between KiSS-1and KiSS-1R mRNA expression was found in the proceeding of HCC invasion and metastasis (P < 0.05).2,The processing of tissue chips was manually modified to construct successfully 8 paraffin blocks of tissue chips in the size of 35mm×25mm×10mm . Every paraffin blocks contained 188 small tissues including HCC, non-neoplastic adjacent liver tissues, non-neoplastic distant liver tissues, normal liver tissues and intrahepatic metastasis lesions. After the tissue chips were stained by H-E, IHC and ISH, the average sheded ratio was 1.40%, 5.92% and 9.76% respectively, and effective rate was 99.58%, 96.25%and 91.19%respectively. Tissue chips were suitable for the histological observation by microscopy and further study.3,The results of ISH and IHC showed that on both mRNA and protein levels, the expression of KiSS-1 in the non-neoplastic adjacent liver tissues, non-neoplastic distant liver tissues or normal liver tissues was significantly higher than that in HCC(P<0.01). With the advance of the pathologic grade and clinical stage of HCC, the expression of KiSS-1 decreased(P<0.05). The expression of KiSS-1 in HCC with metastasis was lower than that in those without metastasis, but it was higher than that in the corresponding metastasis lesions. KiSS-1 expression is not associated with sex, age, tumor size, capsule invasion and histological types(P>0.05). Compared with the the non-neoplastic adjacent liver tissues, non-neoplastic distant liver tissues or normal liver tissues, the expression of MMP-9 in HCCs was significantly increased(P<0.01). The expression of MMP-9 in HCC with metastasis or capsule invasion was higher than that in those without metastasis or capsule invasion, but it was lower than that in the corresponding metastasis lesions. Negative correlation between KiSS-1 and MMP-9 expression was found in the proceeding of HCC invasion and metastasis.Conclusion:1,The modified processing of tissue chips manually was an easy and cost-effective method, decreased the sheded ratio, improved the effective rate and made the tissue chips suitable for the histological observation by microscopy and further study.2,Loss of KiSS-1 expression or overexpression of MMP-9 was closely associated with HCC invasion and metastasis.3,The expression of KiSS-1R mRNA had not associated with all clinicopathologic parameters of HCC.4,For a negative correlation between KiSS-1and KiSS-1R mRNA expression was found in the proceeding of HCC invasion and metastasis , it was suggested that up-regulation of KiSS-1R mRNA expression was a feedback of down-regulation of KiSS-1 mRNA expression. A possible feedback mechanism might exist between KiSS-1and KiSS-1R mRNA expression.5,KiSS-1 expression showed negative correlation with MMP-9 expression in the proceeding of HCC invasion and metastasis on both mRNA and protein levels, so KiSS-1 and MMP-9 expression might interactively co-regulate HCC invasion and metastasis.6,Loss of KiSS-1 expression and overexpression of MMP-9 played important roles in the proceeding of HCC invasion and metastasis and might predict an aggressive clinical behavior and associate with metastatic potential in HCC.
|
PDF Full Text Request