The melanoma differentiation-associated gene mda-6, which encodes the cyclin-dependent kinase inhibitor p21, is differentially expressed during growth, differentiation and progression in human melanoma cells.
The melanoma differentiation-associated gene mda-6, which encodes the cyclin-dependent kinase inhibitor p21, is differentially expressed during growth, differentiation and progression in human melanoma cells.
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发表时间：
1995-05
期刊：
影响因子：
8
通讯作者：
Hongping Jiang;Jian Lin;Zao‐zhong Su;M. Herlyn;Robert S. Kerbel;Bernard E Weissman;Danny R. Welch;Paul B. Fisher
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文献类型：
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作者：
Hongping Jiang;Jian Lin;Zao‐zhong Su;M. Herlyn;Robert S. Kerbel;Bernard E Weissman;Danny R. Welch;Paul B. Fisher
重组人成纤维细胞干扰素(干扰素-β)和抗白血病化合物梅泽莱因(Mezerein)联合诱导人黑色素瘤细胞终末分化，并导致不可逆转的增殖能力丧失。利用消减杂交技术，鉴定出在终末分化和生长受阻的人黑色素瘤细胞中表达增强的cDNA(酱和Fisher，1993；酱等人，1994a)。描述了一种特定的黑色素瘤分化相关(MDA)基因，MDA-6，其表达与黑色素瘤的进展和生长呈负相关。MDA-6与WAF1/CIP1/SDI1相同，WAF1/CIP1/SDI1编码M(R)21,000蛋白(P21)，M(R)21,000蛋白是细胞周期蛋白依赖性激酶的抑制因子。活跃生长的正常黑素细胞、SV40永生化的人类黑素细胞和发育不良的痣细胞系合成高水平的MDA-6mRNA；而活跃增殖的放射状和早期垂直生长阶段的原发黑色素瘤以及转移性人类黑色素瘤细胞产生低水平的MDA-6mRNA。用干扰素-β+MeZ处理原代和转移的人黑色素瘤细胞，可抑制细胞生长，增加MDA-6的表达。当人黑色素瘤细胞生长到高饱和密度或无血清培养时，MDA-6的表达也会增加。使用抗p53和抗p21抗体，发现在生长停滞和分化过程中，p53和p21蛋白水平呈负相关。干扰素-β+MeZ诱导H0-1人黑色素瘤细胞生长停滞和终末分化，导致野生型p53蛋白水平暂时降低，而p21蛋白水平相应增加。在Matrigel辅助的黑色素瘤进展模型中，在早期垂直生长阶段的原代人类黑色素瘤细胞中，MDA-6的表达减少，这些细胞被选为自主或增强裸鼠肿瘤形成的细胞。在转移性人类黑色素瘤细胞中，由于引入正常的人类6号染色体而导致转移潜能丧失，MDA-6mRNA水平增加。综上所述，这些研究表明，MDA-6(P21)可能是黑色素瘤生长、进展和转移的负调控因子。
The combination of recombinant human fibroblast interferon (IFN-beta) and the antileukemic compound mezerein (MEZ) induces terminal differentiation with an irreversible loss of proliferative capacity in human melanoma cells. Using subtraction hybridization, cDNAs were identified that display enhanced expression in terminally differentiated and growth arrested human melanoma cells (Jiang and Fisher, 1993; Jiang et al., 1994a). A specific melanoma differentiation-associated (mda) cDNA, mda-6, is described whose expression inversely correlates with melanoma progression and growth. mda-6 is identical to WAF1/CIP1/SDI1 that encodes the M(r) 21,000 protein (p21) that is an inhibitor of cyclin-dependent kinases. Actively growing normal melanocyte, SV40-immortalized human melanocyte and dysplastic nevus cell lines synthesize elevated levels of mda-6 mRNA; whereas, actively proliferating radial and early vertical growth phase primary melanomas as well as metastatic human melanoma cells produce reduced levels of mda-6 mRNA. Treatment of primary and metastatic human melanoma cells with IFN-beta + MEZ results in growth inhibition and an increase in mda-6 expression. mda-6 expression also increases when human melanoma cells are grown to high saturation densities or when grown in serum-free medium. Using anti-p53 and anti-p21 antibodies, an inverse correlation is found between p53 and p21 protein levels during growth arrest and differentiation. Induction of growth arrest and terminal differentiation in H0-1 human melanoma cells by IFN-beta + MEZ results in a temporal decrease in wild-type p53 protein levels with a corresponding increase in p21 levels. In the Matrigel-assisted melanoma progression model, mda-6 expression decreases in early vertical growth phase primary human melanoma cells selected for autonomous or enhanced tumor formation in nude mice. In metastatic human melanoma cells displaying a loss of metastatic potential resulting from introduction of a normal human chromosome 6, mda-6 mRNA levels increase. Taken together, these studies indicate that mda-6 (p21) may function as a negative regulator of melanoma growth, progression and metastasis.