Isolation of All CD44 Transcripts in Human Epidermis and Regulation of Their Expression by Various Agents

• Published in: PloS one Vol. 11; no. 8; p. e0160952
• Main Authors: Teye, Kwesi, Numata, Sanae, Ishii, Norito, Krol, Rafal P, Tsuchisaka, Atsunari, Hamada, Takahiro, Koga, Hiroshi, Karashima, Tadashi, Ohata, Chika, Tsuruta, Daisuke, Saya, Hideyuki, Haftek, Marek, Hashimoto, Takashi
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 09-08-2016; Public Library of Science (PLoS)
• Subjects: Acetic acid; Acrylamide; Adult; Alternative splicing; Animals; Base Sequence; Biological effects; Biology; Biology and Life Sciences; Calcium; Cancer; Carcinogenesis; CD44 antigen; Cell surface; CHO Cells; Cloning; Cricetinae; Cricetulus; Deoxyribonucleic acid; Dermatology; DNA; Epidermal growth factor; Epidermis; Epidermis - cytology; Epidermis - metabolism; Exons - genetics; Fetal calf serum; Gels; Gene expression; Gene Expression Regulation - drug effects; Genetic aspects; Genetic Variation; Heparan sulfate; Humans; Hyaluronan Receptors - genetics; Hydrogen; Hydrogen peroxide; Isoforms; Keratinocytes; Keratinocytes - cytology; Keratinocytes - metabolism; Keratinocytes - pathology; Medicine; Medicine and Health Sciences; Metastasis; Nucleotide sequence; Pathogenesis; Phorbol 12-myristate 13-acetate; Physiological aspects; Polymerase chain reaction; Protein Isoforms - genetics; Proteoglycans; Psoriasis; Research and Analysis Methods; Retinoic acid; RNA, Messenger - genetics; RNA, Messenger - isolation & purification; Skin Neoplasms - pathology; Tumorigenesis; Japan; New York; United States > US
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0160952

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.