파라핀 절편의 변성된 단백질을 분석하기 위한 ImmunoMemBlot 방법의 적용 -법랑모
In order to perform the protein analysis using the paraffin sections previous fixed with formalin, we applied theImmunoMemBlot (IMB) method1) to detect the epitopes of target proteins with specific antibodies. In this study the proteinextracts were obtained from the paraffin sections of each representative case of ameloblastoma, adenomatoid odontogenictumor (AOT), and normal gingiva, and more a protein extract from fresh tissue of ameloblastoma was also compared toevaluate the IMB results used with 24 different antibodies. First of all, in the comparison between the paraffin section extractand fresh tissue extract of ameloblastoma, the latter consistently showed more positive IMB reaction than the former.Meanwhile, the paraffin section extract of ameloblastoma was more comparable with that of normal gingival, disclosing thatmost of proliferating genes, oncogenes, and apoptosis related genes, i.e., PCNA, CDK4, c-erbB2, CEA, p53, Bax, Bad, FLIP,FAS, Bcl-2, p21, N-ras, MMP-2, MMP-9, caspase-3, -8, -9, were highly expressed in ameloblastoma, but EGFR, HGF, andVEGF were similarly expressed both in the ameloblastoma and in normal human gingiva. On the other hand, the comparisonbetween ameloblastoma and AOT both in the immunohistochemistry and IMB using their paraffin section extracts clearlydemonstrated that the ameloblastoma showed more expression of proliferating genes and oncogenes while the AOT showedmore expression of apoptosis related genes, i.e., Bax, Bad, FLIP, and caspase-9. Taken together, these data suggest thatthe IMB can be used for the primary screening of quantitative protein analysis using the paraffin section extract, and thatthe IMB results could be evaluated in conjunction with the immunohistochemical observation.