APPLICATION OF MICROWAVE(MW)-FIXED TISSUE SAMPLES TO IN SITU HYBRIDIZATION: DETECTION OF CARCINOEMBRYONIC ANTIGEN (CEA) mRNA AND ONCOGENE TRANSCRIPTS IN MW-FIXED COLORECTAL CARCINOMAS USING SULFONATED PROBES (<Symposium>In situ Hybridization I. : Methodol
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概要
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Microwave (MW) energy has been reported to permit rapid fixation and good conservation of tissue antigens as well as of tissue morphology. In this paper, we examined the applicability of MW-fixed tissue specimens to in situ hybridization. The preliminary analysis by Northern blot hybridization revealed that no degradation of mRNA occurred following MW irradiation to tissue samples. To determine the optimal conditions for in situ hybridization of MW-fixed tissue sections,we first investigated the in situ localization of carcinoembryonic antigen (CEA) mRNA in MW-fixed colorectal carcinomas using a sulfonated cDNA probe. The results indicated that pretreatment of sections with HCl could be omitted and the concentration of proteinase K for tissue digestion could be reduced in MW-fixed specimens. Thus, these specimens provided stronger CEA mRNA signals when compared with routinely fixed ones. On the basis of this result, we examined oncogene expression in 26 colorectal cancer specimens fixed by MW irradiation, and successfully detected Ha-ras mRNA in 6 (23%), Ki-ras mRNA in 7 (27%), and c-myc mRNA in 12 (46%) specimens. The results of in situ hybridization for the oncogene mRNAs agreed well with those obtained by the immumohistochemical analysis of oncogene products using the specific antibodies to p21^<ras> and p62^<myc>. Our observations indicated the usefulness of rapid MW fixation for the preservation of mRNA in tissue preparations. MW-fixed tissue samples are applicable to in situ hybridization as well as to the immunohistochemical analysis of the expression of various genes.
- 日本組織細胞化学会の論文
著者
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Higashiyama M
Department Of Thoracic Surgery Osaka Medical Center For Cancer And Cardiovascular Diseases
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Higashiyama Masahiko
大阪府立成人病センター研究所
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Higashiyama Masahiko
The Second Department Of Surgery Osaka University Medical School
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Mori T
Kyoto Univ. Kyoto Jpn
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Monden Takushi
The Second Department Of Surgery Osaka University Medical School
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Mori Takesada
The Second Department Of Surgery Osaka University Medical School
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MUROTANI MASAHIRO
The Second Department of Surgery, Osaka University Medical School
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MORIMOTO HIDEKI
The Second Department of Surgery, Osaka University Medical School
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KAWASAKI YASUHITO
The Second Department of Surgery, Osaka University Medical School
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SHIMANO TAKASHI
The Second Department of Surgery, Osaka University Medical School
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Murotani Masahiro
The Second Department Of Surgery Osaka University Medical School
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Morimoto H
Hiroshima Univ. Fac. Dentistry Hiroshima Jpn
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Shimano Takashi
The Second Department Of Surgery Osaka University Medical School
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Higashiyama Masahiko
Department Of Thoracic Surgery Osaka Medical Center For Cancer And Cardiovascular Diseases
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Kawasaki Yasuhito
The Second Department Of Surgery Osaka University Medical School
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