Higashide T, Hinoda Y, Itoh J, Takahashi H, Satoh Y, Ibayashi Y, Imai K, Yachi A
Department of Internal Medicine (Section 1), Sapporo Medical College.
Jpn J Cancer Res. 1990 Nov;81(11):1149-54. doi: 10.1111/j.1349-7006.1990.tb02527.x.
Expression of mRNAs of carcinoembryonic antigen (CEA) and nonspecific cross-reacting antigen (NCA) genes in colorectal carcinomas and adenomas was investigated by in situ hybridization (ISH) with specific biotinylated probes. Hybridization was clearly detected throughout the cytoplasm of 7 out of 15 adenomas and 13 out of 15 carcinomas with the CEA cDNA probe, and in 6 out of 15 adenomas and 10 out of 15 carcinomas with the NCA cDNA probe. The intensity of signal appeared to be stronger in carcinomas than that in adenomas, and the CEA and NCA mRNAs were expressed together in most of the positive tissue specimens. On the other hand, noninvaded tissues adjacent to the carcinoma did not show any signal except for 4 cases faintly stained with the NCA probe. This finding was partly confirmed by Northern blot analysis which indicated that the specific bands for the CEA and NCA mRNAs were more intense in RNAs from carcinoma tissues than in those from adjacent noninvaded tissues. These data suggest that the ISH technique with biotinylated probes could be of use for analyzing expression and localization of CEA and related genes on tissue sections.
采用特异性生物素化探针原位杂交(ISH)技术,研究结直肠癌和腺瘤中癌胚抗原(CEA)和非特异性交叉反应抗原(NCA)基因的mRNA表达情况。使用CEA cDNA探针时,在15个腺瘤中的7个以及15个癌中的13个的整个细胞质中均清晰检测到杂交信号;使用NCA cDNA探针时,在15个腺瘤中的6个以及15个癌中的10个的整个细胞质中均清晰检测到杂交信号。癌组织中的信号强度似乎比腺瘤中的更强,并且在大多数阳性组织标本中,CEA和NCA mRNA共同表达。另一方面,癌旁未受侵犯的组织除4例被NCA探针轻微染色外,未显示任何信号。Northern印迹分析部分证实了这一发现,该分析表明,癌组织RNA中CEA和NCA mRNA的特异性条带比相邻未受侵犯组织RNA中的更强烈。这些数据表明,使用生物素化探针的ISH技术可用于分析组织切片上CEA及相关基因的表达和定位。