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Isolation of Mouse Epidermal Keratinocytes and Their In Vitro Clonogenic Culture
06:16

Isolation of Mouse Epidermal Keratinocytes and Their In Vitro Clonogenic Culture

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Gene expression changes associated with altered growth and differentiation in benzo[a]pyrene or arsenic exposed

Damon S Perez1, Robert J Handa, Raymond S H Yang

  • 1Quantitative and Computational Toxicology Group, Center for Environmental Toxicology and Technology, Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA. perez.damon@mayo.edu <perez.damon@mayo.edu>

Journal of Applied Toxicology : JAT
|September 20, 2007
PubMed

Insights

Arsenic and benzo[a]pyrene (BaP) alter human skin cell growth and differentiation by affecting different genes. These toxic chemicals impact molecular pathways controlling keratinocyte proliferation and differentiation.

Area of Science:

  • Toxicology
  • Molecular Biology
  • Dermatology

Background:

  • Arsenic and benzo[a]pyrene (BaP) are environmental toxins known to inhibit terminal differentiation and alter growth potential in normal human epidermal keratinocytes (NHEK) in vitro.
  • Understanding the molecular mechanisms underlying these cellular changes is crucial for assessing toxicological risks and developing preventative strategies.

Purpose of the Study:

  • To identify molecular alterations in normal human epidermal keratinocytes (NHEK) induced by arsenic and benzo[a]pyrene (BaP).
  • To compare the gene expression profiles and identify specific molecular pathways affected by each toxicant.

Main Methods:

  • Gene expression profiling using microarray analysis on NHEK treated with BaP or arsenic.
  • Quantitative real-time RT-PCR to validate microarray findings for selected genes involved in keratinocyte growth and differentiation.

Main Results:

  • BaP (2.0 microm) induced 85 genes and suppressed 17; Arsenic (5.0 microm) induced 106 genes and suppressed 15.
  • Real-time RT-PCR confirmed BaP increased mRNA levels of AIBP63, IFIT5, IL1A, IL1B, and RASGRP1.
  • Real-time RT-PCR confirmed arsenic increased mRNA levels of RB1, ARID4A, TSC22D1, MNT, and RAD50.

Conclusions:

  • Arsenic and BaP target distinct genes and molecular pathways in NHEK.
  • These findings provide molecular markers for chemical-specific effects on keratinocyte proliferation and differentiation.
  • Further mechanistic studies can correlate these molecular alterations with observed differentiation defects.

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