Isolation of All CD44 Transcripts in Human Epidermis and Regulation of Their Expression by Various Agents.
Isolation of All CD44 Transcripts in Human Epidermis and Regulation of Their Expression by Various Agents.
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DOI：
10.1371/journal.pone.0160952
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Hashimoto T
中科院分区：
文献类型：
--
作者：
Teye K;Numata S;Ishii N;Krol RP;Tsuchisaka A;Hamada T;Koga H;Karashima T;Ohata C;Tsuruta D;Saya H;Haftek M;Hashimoto T
CD 44是一种细胞表面蛋白多糖，参与许多生物学事件。CD 44转录物经历复杂的选择性剪接，导致许多功能不同的同种型。然而，迄今为止，这些亚型在人表皮中的性质尚未得到充分确定。在这项研究中，我们从正常人表皮中分离出所有的CD 44转录本，并研究它们的表达是如何调节的。通过RT-PCR方法，我们发现人表皮中存在多种不同的CD 44转录本，并通过克隆的方法从琼脂糖和丙烯酰胺凝胶中的DNA条带中获得了所有这些转录本。详细的序列分析显示了18个CD 44转录本，其中3个是新的。接下来，我们研究了10种不同药物对培养的人角质形成细胞中CD 44转录物表达的影响，发现几种药物，特别是表皮生长因子，过氧化氢，佛波醇12-肉豆蔻酸酯13-乙酸酯，视黄酸，钙和胎牛血清以不同的模式调节其表达。此外，发现正常和恶性角质形成细胞在血清刺激和随后的饥饿后产生不同的CD 44转录物，这表明特定的CD 44亚型通过不同的CD 44介导的生物学途径参与肿瘤发生。
CD44, a cell surface proteoglycan, is involved in many biological events. CD44 transcripts undergo complex alternative splicing, resulting in many functionally distinct isoforms. To date, however, the nature of these isoforms in human epidermis has not been adequately determined. In this study, we isolated all CD44 transcripts from normal human epidermis, and studied how their expressions are regulated. By RT-PCR, we found that a number of different CD44 transcripts were expressed in human epidermis, and we obtained all these transcripts from DNA bands in agarose and acrylamide gels by cloning. Detailed sequence analysis revealed 18 CD44 transcripts, 3 of which were novel. Next, we examined effects of 10 different agents on the expression of CD44 transcripts in cultured human keratinocytes, and found that several agents, particularly epidermal growth factor, hydrogen peroxide, phorbol 12-myristate 13-acetate, retinoic acid, calcium and fetal calf serum differently regulated their expressions in various patterns. Furthermore, normal and malignant keratinocytes were found to produce different CD44 transcripts upon serum stimulation and subsequent starvation, suggesting that specific CD44 isoforms are involved in tumorigenesis via different CD44-mediated biological pathways.