The Expression And Interaction Of CD147and ABCG2in Epidermis Of Psoriasis And Hacat Cells

AbjectiveThe Extracellular Matrix Metalloproteinase Inducer (EMMPRIN, CD147, basigin), an immunoglobulin-like receptor, is an integral plasma membrane glycoprotein which is enriched on the surface of many cancer cells. The expression of CD147on psoriasis keratinocytes has been known to promote the development of psoriasis. ABCG2(breast cancer resistance protein, BCRP/ABCG2) is a member of the G-subfamiliy of the ATP-binding cassette (ABC)-transporter superfamily. This half-transporter is assumed to function as an important pump limiting cellular accumulation of various compounds. Though it has been referred in many reports to promote cell proliferation, the location and regulation of ABCG2in psoriasis is still under-investigated. We did our present experiment to study the expression and the regulation of CD147and ABCG2in epidermis of psoriasis and HaCaT cells.Methods(1) The location and expression of CD147and ABCG2in the keratinocytes of psoriasis were detected by immunohistochemistry;(2) The location and expression of CD147and ABCG2in the epidermis of psoriasis were detected by laser scanning confocal microscope combined with immunohistochemical double labeling;(3) Establish a stable CD147-overexpression HaCaT cells (HaCaT-C) and ABCG2-overexpression HaCaT cells (HaCaT-A),which were defined by Western blot;(4) The complex of CD147and ABCG2were detected by IP (immunoprecipitation, IP) in HaCaT-A;(5) We established ABCG2and CD147-cooverexpression HEK293FT cells to analyze the binding sites of CD147and ABCG2by IP;(6) The expression of ABCG2induced by MX were detected in HaCaT-C and HaCaT-N cell line by Western blot;(7) Alarm Blue assay was carried out to show the difference of chemosensitivity to MX between HaCaT-C cells and HaCaT-N cells;(8) The expression of CD147and ABCG2were detected in HaCaT cells treated with TNF-a by Western blot.Results(1) Campared with normal skin, CD147and ABCG2were located in the whole layers of the epidermis of psoriasis and mainly expressed on the membrance of keratinocytes;(2) CD147and ABCG2were colocated on the membrance of keratinocytes in psoriasis;(3) We established a stable cell line HaCaT-C with overexpression CD147;(4) CD147and ABCG2formed a complex in HaCaT-A detected by IP;(5) The binding sites of CD147and ABCG2were located on the transmembrane domain of CD147;(6) The induced-expression of ABCG2by MX were1.56±0.16fold in HaCaT-C compared with HaCaT-N by Western blot;(7) Alarm Blue assay showed a decrease MX cytoxicity to HaCaT-C than HaCaT-N cells; (8) The expression of CD147and ABCG2were higher in HaCaT cells treated with TNF-α.Conclusion(1) Campared with normal skin, CD147and ABCG2were located in the whole layers of the epidermis of psoriasis and mainly expressed on the membrance of keratinocytes.(2) CD147and ABCG2formed a complex and the binding site was located on the transmembrane domain of CD147.(3) CD147played a role in the regulation of the expression and fuction of ABCG2.