A Preliminary Study On Glycolysis In Keratinocytes Of Psoriasis Vulgaris And HaCaT Cell Line

Objective:Psoriasis vulgaris is a chronic inflammatory dermatological disease, which is characterized by hyper-proliferation and parakeratosis in epidermis, and telangiectasisa and inflammation in dermis. There are same characteristics between psoriasis and malignant tumors:hyper-proliferation and local hypoxia. Hypoxia-inducible factor-la (HIF-la) directly regulates most enzymes necessary for glycolysis, so it shifts energy production by increasing glycolysis. In preliminary studies we found that CD147was highly expressed in psoriatic patients’ skin lesions and peripheral blood cells, and a susceptibility gene of psoriasis. We also demonstrated that CD147played a vital role in glycolysis in melanoma, and could be regulated by HIF-1α and mediated melanoma cells’metastasis and invasion. In this study, we aimed to explore the relationship among HIF-1α, CD147and proteins associated with glycolysis as well as roles that they played in keratinocytes of psoriasis vulgaris and HaCaT cells. To further provide a new theoretical basis for psoriasis pathogenesis.Methods:1.The location and expressions of HIF-1α, CD147, GLUT1, HK2and PKM2were detected in both psoriasis vulgaris lesions and normal epidermis by immunohistochemistry, followed by statistic analysis;2. Established small interfering RNAs(siRNA) and transfected them into HaCaT cells to knock down HIF-1α, detected protein expressions of HIF-la, CD147, GLUT1, HK2and PKM2by Western Blot; 3. Added CoCl2in HaCaT cells to induce chemical hypoxia, detected protein expressions of HIF-1α, CD147, GLUT1, HK2and PKM2by Western Blot;4. Defined location and expressions of CD147and GLUT1in psoriasis epidermis by fluorescent double labeling;5. Transfected siRNAs and CD147full length plasmids into HaCaT cells to knock down and overexpress CD147respectively, detected protein expressions of CD147and GLUT1by Western Blot;6. Transfected siRNAs and GLUT1full length plasmids into HaCaT cells to knock down and overexpress GLUT1respectively, detected protein expressions of GLUT land CD147by Western Blot.Results:1. HIF-1α, CD147, GLUT1, HK2and PKM2expressed significantly higher in psoriasis vulgaris lesions than in normal epidermis (P<0.05);2. The expression of CD147, GLUT1and HIF-1α positively correlated with one another in psoriasis lesions (P<0.05), and expression of HK2and PKM2had no correlation with the others(P>0.05);3. After knocking down HIF-1α in HaCaT cells, protein expressions of HIF-1α, CD147, GLUT1and HK2were significantly decreased (P <0.05), but no significant change in PKM2’s expression (P>0.05);4. After hypoxia by adding CoCl2, protein expressions of HIF-1α, CD147, GLUT1and HK2were significantly increased (P<0.05), but no significant change in PKM2’expression (P>0.05).5. CD147and GLUT1co-localized in basal cell membranes and partial stratum spinosum cell membranes of psoriasis lesional epidermis;6. After knocking down and overexpressing CD147in HaCaT cells, no significant change observed in GLUTl’s expression (P>0.05);7. After knocking down and overexpressing GLUT1in HaCaT cells, no significant change observed in CD147’s expression (P>0.05). Conclusion:1. Glycolytic ability probably increases in keratinocytes of psoriasis vulgaris;2. Hypoxia in HaCaT cells leads to up-regulate HIF-la and futher increase expressions of CD147, GLUT1and HK2, that may contribute to increased glycolytic ability in HaCaT cells.