Search for oncogenic regulators in an autocrine tumor model using differential display PCR: Identification of novel candidate genes including the calcium channel mtrp6
Search for oncogenic regulators in an autocrine tumor model using differential display PCR: Identification of novel candidate genes including the calcium channel mtrp6
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DOI：
10.1038/sj.onc.1202445
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发表时间：
1999-02-18
期刊：
影响因子：
8
通讯作者：
Moroni, C
中科院分区：
文献类型：
--
作者：
Buess, M;Engler, O;Moroni, C
之前已经建立了造血多步肿瘤模型，其中IL-3依赖性PB-3c肥大细胞在体内表达v-H-ras后进展为产生IL-3的自分泌肿瘤。这种致癌进程的核心是隐性步骤，该步骤可通过细胞融合逆转，并导致 IL-3 mRNA 稳定，同时激活自分泌环。通过差异显示 PCR 比较 IL-3 依赖性 PB-3c 和 IL-3 自分泌 V2D1 肿瘤细胞，发现肿瘤中有 12 个差异表达基因，其中 8 个上调，4 个下调。它们包括四种蛋白酶（小鼠肥大细胞蛋白酶 2、颗粒酶 B、胃蛋白酶原 F 和丝氨酸蛋白酶 1）和两种代谢酶（腺嘌呤磷酸核糖基转移酶和果糖 1,6-二磷酸酶）。为了验证，在独立的 PB-3c 前体克隆及其肿瘤衍生物中测试了已识别基因的表达。内源性逆转录病毒 IAP 元件和三种未知转录物的表达在所有肿瘤系中一致上调。在体细胞杂交体中，其中两种未知的 cDNA 显示显性表达模式，另一种显示隐性表达模式。一种在前体中表达但在肿瘤细胞中下调的转录本被克隆并鉴定为鼠钙通道 mtrp6。
A hemopoietic multistep tumor model, in which IL-3 dependent PB-3c mast cells, following expression of v-H-ras progress in vivo to IL-3 producing autocrine tumors has previously been established. Central for this oncogenic progression is a recessive step, which is reversible by cell fusion and leads to stabilization of IL-3 mRNA with concomitant activation of the autocrine loop. Comparing the IL-3 dependent PB-3c and the IL-3 autocrine V2D1 tumor cells with differential display PCR revealed 12 differentially expressed genes of which eight were upregulated and four downregulated in the tumor. They included four proteases (mouse mast cell protease 2, granzyme B, pepsinogen F and serine protease 1) and two metabolic enzymes (adenine phosphoribosyltransferase and fructose1,6-bisphosphatase). For validation, expression of the identified genes was tested in independent PB-3c precursor clones and their tumor derivatives. Expression of an endogenous retroviral IAP element and three unknown transcripts were consistently upregulated in all tumor lines. In somatic cell hybrids, two of these unknown cDNAs showed a dominant and one a recessive expression pattern. One transcript, expressed in the precursor but downregulated in the tumor cells, was cloned and identified as the murine calcium channel mtrp6.