Gene expression profiling of human epidermal keratinocytes in simulated microgravity and recovery cultures.
Simulated microgravity (SMG) bioreactors and DNA microarray technology are powerful tools to identify "space genes" that play key roles in cellular response to microgravity. We applied these biotechnology tools to investigate SMG and post-SMG recovery effects on human epidermal keratinocytes by exposing cells to SMG for 3, 4, 9, and 10 d using the high aspect ratio vessel bioreactor followed by recovery culturing for 15, 50, and 60 d in normal gravity. As a result, we identified 162 differentially expressed genes, 32 of which were "center genes" that were most consistently affected in the time course experiments. Eleven of the center genes were from the integrated stress response pathways and were coordinately down-regulated. Another seven of the center genes, which are all metallothionein MT-I and MT-II isoforms, were coordinately up-regulated. In addition, HLA-G, a key gene in cellular immune response suppression, was found to be significantly up-regulated during the recovery phase. Overall, more than 80% of the differentially expressed genes from the shorter exposures (<or=4 d) recovered in 15 d; for longer (>or=9 d) exposures, more than 50 d were needed to recover to the impact level of shorter exposures. The data indicated that shorter SMG exposure duration would lead to quicker and more complete recovery from the microgravity effect.
Clement
Jade Q
JQ
Department of Chemistry and NASA University Research Center for Biotechnology and Environmental Health, Texas Southern University, Houston, TX 77004, USA. clement_jq@tsu.edu
Lacy
Shareen M
SM
Wilson
Bobby L
BL
D019149:Bioreactors; D015152:Blotting, Northern; D018929:Cell Culture Techniques; D002460:Cell Line; D016000:Cluster Analysis; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006112:Gravitation; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D013997:Time Factors; D018474:Weightlessness Simulation