Expression and up-regulation of alternatively spliced transcripts of melastatin, a melanoma metastasis-related gene, in human melanoma cells
Expression and up-regulation of alternatively spliced transcripts of melastatin, a melanoma metastasis-related gene, in human melanoma cells
复制标题
DOI：
10.1006/bbrc.2000.3894
复制
发表时间：
2000-12-09
影响因子：
3.1
通讯作者：
Setaluri, V
中科院分区：
文献类型：
--
作者：
Fang, D;Setaluri, V
一种新的转移抑制因子Melastatnn(MLSN1)的表达缺失最近被报道与黑色素瘤细胞的转移潜能有关。利用差异显示分析，我们在用分化诱导剂六亚甲基双乙酰胺(HMBA)处理的色素转移性人黑色素瘤细胞中鉴定出MLSN1过表达的基因。在这项研究中，我们发现MLSN1的多个短转录本存在于黑素细胞和转移性黑色素瘤细胞系中，而全长5.4-kb的mRNA只有在黑素细胞中才能检测到。用分化诱导剂HMBA处理色素黑色素瘤细胞，会导致5.4kb的MLSN1mRNA和短RNA的上调。对一组无色素的原发和转移性黑色素瘤细胞系的分析显示，在一些黑色素瘤细胞系中1.8kb的mRNA表达很弱。用与MLSN1mRNA全长不同区域对应的DNA探针和寡核苷酸引物进行Northern印迹和RT-PCR分析表明，这些短转录本含有主要对应于5.4kb基因5‘端或S’端的序列。HMBA似乎上调了主要来自5‘端的MLSN1转录本。CAMP和蛋白激酶C通路的调节剂对MLSN1的表达无明显影响。我们的数据显示，在培养的色素黑色素瘤细胞中存在多种MLSN1转录本，既有成分表达的，也有诱导的，这表明MLSN1的表达可以在转录和mRNA处理水平上进行调节，(C)2000学术出版社。
Loss of expression of a novel suppressor of metastasis, melastatnn (MLSN1), has recently been reported to correlate with metastatic potential of melanoma cells. Using differential display analysis, we identified MLSN1 among genes overexpressed in pigmented metastatic human melanoma cells treated with the differentiation inducer hexamethylene bisacetamide (HMBA). In this study, we show that multiple short transcripts of MLSN1 are present in melanocytes and pigmented metastatic melanoma cell lines while the full-length 5.4-kb mRNA is detectable only in melanocytes. Treatment of pigmented melanoma cells with the differentiation-inducing agent, HMBA, results in up-regulation of the 5.4-kb MLSN1 mRNA as well as short RNAs. Analysis of a panel of nonpigmented primary and metastatic melanoma cell lines showed weak expression of a 1.8-kb mRNA in a few melanoma cell lines. Northern blot and RT-PCR analyses with DNA probes and oligonucleotide primers that correspond to distinct regions of full-length MLSN1 mRNA indicated that the short transcripts contained sequences corresponding primarily to either 5'- or S'-end of the 5.4-kb mRNA. HMBA appears to up-regulate MLSN1 transcripts derived mainly from the 5'-end. Modulators of cAMP and protein kinase C pathways had no significant effect on MLSN1 expression. Our data show that multiple MLSN1 transcripts, both constitutively expressed and inducible, are present in cultured pigmented melanoma cells, and suggest that MLSN1 expression can be regulated at the level of both transcription and mRNA processing, (C) 2000 Academic Press.