我們研究室以Kinase activity assay和western blot分析人類肝癌組織之PKC，結果發現Kinase總活性上昇，PKCα isoform的量顯著下降，而PKCζ& PKCδ則是顯著上昇，顯示PKC異構型酵素在癌瘤發展上扮演重要的角色，所以本計畫中同時以Specific PKC isoform antibodies進行Immnohistochemistry分析，以獲知接種S-180癌瘤細胞之老鼠個臟器之Cytosolic CAI，CAII及CAIII蛋白表現量變化與PKC isoforms變化是否相關？顯果顯示：(1)以Immunoblot分析tissues中Cytosolic CAI，CALL及CAIII蛋白表現量變化得知其變化量與Tumor progression的程度有相關性(2)以Enzyme assays分析Tissues中CA活性得知CA活性隨著CAI，CAII及CAIII蛋白量的變化而改變(3)以Immnohistochemistry分析組織中CAI，CAII及CAIII獲知Tissues中CAI，CAII及CAIII細胞分佈情形及變化(4)以Immnohistochemistry組織中PKC isoforms的變化，獲知PKC蛋白表現量變化關鍵性角色。
The expressions of CAI and CAII have been most frequently investigated in a variety of tumor cells and cell lines, but it has been difficult to find a clear-cut relationship between the expression of CA isoenzymes in normal and malignant cell. However, no evidence of a direct relationship between normal and malignancy. CA expression has been presented for CAs I through VII. It appears that only the recently characterized isoform CAIX and CAXII are exceptional, as it expression can be associated with tumorigenesis. We also found that the level of membrane-bound PKCα in the cancer tissue significantly decreased as compared with that in the adjacent normal tissue, whereas there was no alternation in the cytosolic PKCα. In addition, there was a significant correlation between the level of PKCα and the tumor size. The levels of PKCζand PKCδ significantly increased in both cytosolic and membrane. In this project, we examined the protein and the messenger RNA (mRNA) levels of cytosolic CAs (including CAI, CAII and CAIII) that was correlated to PKC expression in the organs of S-180 bearing mice. We also examined the protein expression of cytosolic CA by immunohistchemical methods. The possibility that cytosolic CA serving as an indicator for a favorable treatment outcome in the organs of S-180 bearing mice were obtained.
