Fleming M G, Howe S F, Graf L H
Department of Dermatology, Rush-Presbyterian-St. Luke's Medical Center, Chicago, IL 60612-3833.
Am J Dermatopathol. 1994 Aug;16(4):383-91. doi: 10.1097/00000372-199408000-00005.
Expression of IGF-I mRNA and protein was evaluated in pigmented lesions by in situ hybridization and immunohistochemistry. An IGF-I cDNA clone (phigf1) was subcloned into pBluescript KS II-. Both sense and antisense 35S riboprobes were prepared and used for in situ hybridization on formalin-fixed, paraffin-embedded specimens. Control hybridizations with a beta-actin probe were also performed. Grains were counted in 787-microns2 melanocytic areas of sections hybridized with the antisense IGF-I probe. Seven common nevi contained a mean of 218 grains; nine dysplastic nevi, a mean of 463 grains; eight early primary melanomas, a mean of 402 grains; five advanced primary melanomas, a mean of 217 grains; and nine metastatic melanomas, a mean of 194 grains. The differences between common and dysplastic nevus, common nevus and early melanoma, early and advanced primary melanoma, and early primary melanoma and metastatic melanoma were statistically significant. Keratinocytes also expressed abundant IGF-I message. IGF-I protein was demonstrable by immunohistochemistry in melanocytes and keratinocytes. These results suggest that progression-associated variation occurs in the net expression of IGF-I mRNA in melanocytic tumors.
通过原位杂交和免疫组织化学方法，在色素沉着病变中评估IGF-I mRNA和蛋白的表达。将IGF-I cDNA克隆（phigf1）亚克隆到pBluescript KS II-中。制备正义和反义35S核糖探针，并用于福尔马林固定、石蜡包埋标本的原位杂交。还用β-肌动蛋白探针进行对照杂交。在与反义IGF-I探针杂交的切片的787平方微米黑素细胞区域中计数颗粒。七个普通痣平均含有218个颗粒；九个发育异常痣平均含有463个颗粒；八个早期原发性黑色素瘤平均含有402个颗粒；五个晚期原发性黑色素瘤平均含有217个颗粒；九个转移性黑色素瘤平均含有194个颗粒。普通痣与发育异常痣、普通痣与早期黑色素瘤、早期与晚期原发性黑色素瘤以及早期原发性黑色素瘤与转移性黑色素瘤之间的差异具有统计学意义。角质形成细胞也表达丰富的IGF-I信息。通过免疫组织化学可在黑素细胞和角质形成细胞中检测到IGF-I蛋白。这些结果表明，黑素细胞肿瘤中IGF-I mRNA的净表达存在与进展相关的变化。
