Francia G, Mitchell S D, Moss S E, Hanby A M, Marshall J F, Hart I R
Richard Dimbleby Department of Cancer Research, Rayne Institute, United Medical School, St. Thomas' Hospital, London, United Kingdom.
Cancer Res. 1996 Sep 1;56(17):3855-8.
To identify genes involved in the melanocyte to malignant melanoma conversion, we have applied differential display to the comparison of syngeneic murine B16F10 (metastatic melanoma) and Melan-a-immortalized melanocyte cell lines. Approximately 7000 bands were analyzed, revealing approximately 80 to be differentially displayed. Reverse Northern blotting and subsequent Northern blotting confirmed the reproducible differential expression of four transcripts. Three B16F10-specific bands encode novel genes or partially sequenced cDNAs of unknown function. One Melan-a-specific band was found to be identical to the 3' end region of the mouse Annexin VI mRNA and shown to have a reduced message expression in B16F10 relative to Melan-a. Differential expression was confirmed at the protein level with Western blotting using a rabbit polyclonal antiserum. Immunohistochemistry of human melanoma specimens with this antiserum revealed a decrease or loss of Annexin VI expression as melanomas progressed from a benign to a more malignant phenotype. Our results provide further evidence for a potential role of Annexin VI in tumor suppression.
为了鉴定参与黑素细胞向恶性黑色素瘤转化的基因，我们应用差异显示技术比较了同基因小鼠B16F10（转移性黑色素瘤）和Melan - a永生化黑素细胞系。分析了大约7000条条带，发现约80条存在差异显示。反向Northern印迹和随后的Northern印迹证实了4种转录本可重复的差异表达。3条B16F10特异性条带编码新基因或功能未知的部分测序cDNA。发现1条Melan - a特异性条带与小鼠膜联蛋白VI mRNA的3'末端区域相同，并且相对于Melan - a，在B16F10中其信使表达降低。使用兔多克隆抗血清进行的蛋白质印迹在蛋白质水平上证实了差异表达。用该抗血清对人黑色素瘤标本进行免疫组织化学分析显示，随着黑色素瘤从良性表型发展为更恶性的表型，膜联蛋白VI表达降低或缺失。我们的结果为膜联蛋白VI在肿瘤抑制中的潜在作用提供了进一步的证据。
