Gene Expression Profiles Analysis Of HaCaT Kerationocytes During Interaction With Cryptococcus Neoformans In Vitro

Objective:1.To observe the change of morphology, adhesion and integration with light microscope when cryptococcus neoformans coculturing with HaCaT keratinocytes for 2 hour,4 hour,12 hour and 24 hour in vitro.2.To learn about the interaction of cryptococcus neoformans and HaCaT keratinocytes with gene expression profiles microarray in vitro.Methods: 1. HaCaT keratinocytes were subcultured for 2 to 3 times and modulated the cells concentration to 1×106-7/ ml when it approaches 95% fusion condition,then cocultured in incubator at 37℃and 5% CO2 with l×107-8/mL yeast suspension or fresh cell culture fluids, and oberved with phrase-contrast microscope at different time points.2.Detect the change of gene expression profiles in experimental group and contrast group with homo sapiens genome expression profiles microarray. Results: 1.We could see the adhesion and anastomosis of cryptococcus neoformans and HaCaT keratinocytes,but there was no apparent splitting or fracture phenomenon.2.We found 138 genes up-regulated in expression level when the experimental group compared to the control group,and of which 127 genes was known to us while 11 genes was unkonwn.They involved in cell adhesion and endocytosis, apoptosis, transcription regulation,signal transduction, protein amino acid phosphorylation and many other biological processes,and maybe related to the invasion of cryptococcus neoformans to HaCaT keratinocytes.Discussion: HaCaT keratinocytes could adhere to and swallow cryptococcus neoformans during coculturing in vitro.This maybe caused by the up-regulated gene expression of some genes and at the same time HaCaT keratinocytes increased the apoptosis of itself.So we presumed that the process of adhesion, endocytosis and apoptosis of HaCaT keratinocytes during the invasion of cryptococcus neoformans was the probable pathogenesis of cutaneous cryptococcosis.