Gene transfection in human keratinocyte and melanocyte

1首先，我们研究了ha标记的人凝胶蛋白cDNA转染人鳞癌细胞系HEC46和人黑色素瘤细胞系，代替角化细胞和黑素细胞。我们以人类黑色素瘤组织和正常皮肤组织作为对照，在Western blotting中发现了一种独特的85Kda蛋白通过使用抗gelsolin抗体对38个黑色素瘤样本进行gelsolin表达的Western blotting分析，我们发现了一个缺乏c -末端结构域的gelsolin p85 （GSNp85）的新截断变体，该变体总是与野生型gelsolin共表达。与放射状生长期（RGP）黑色素瘤相比，GSNp85变体在垂直生长期（VGP）黑色素瘤中表达更为频繁，在其他皮肤癌、正常皮肤组织和痣组织中不表达，除了2种先天性交界处类型。这些数据提示，GSNp85在黑色素瘤中的出现可能是肿瘤晚期的新指标。

1 At first, we studied the transfection of ha-tagged human gelsolin cDNA in human squamous carcinomacell line HEC46 and human melanoma cell line in stead of keratinocyte and melanocyte. We used human melanoma tissue and normal skin tissue as a control, and found out a unique 85Kda protein in Western blotting.2 By Western blotting analysis of 38 melanoma samples for the expression of gelsolin using anti-gelsolin antibodies, we identified a new truncated variant of gelsolin p85 (GSNp85) lacking the C-terminal domain, which was always co-expressed with wild type gelsolin. The GSNp85 variant was more frequently expressed in vertical growth phase (VGP) melanomas as compared to radial growth phase (RGP) melanomas and was not expressed in other skin cancers, normal skin tissues, and nevi tissues except for 2 congenital junctional types. These data suggest that the occurrence of GSNp85 in melanoma may be a new indicator for the advanced stage of the tumor.