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Microarray Analysis of Gene Expression in Metolachlor Treated Human Liver Cells
Presentation Type
Oral Presentation (Electronic Copy Not Available)
Keywords
Metolachlor--Physiological effect; Liver--Effect of chemicals on;
Abstract
Metolachlor, a commonly used pre-emergent pesticide in the Midwest, is a frequent contaminant of ground and surface waters. It inhibits protein and chlorophyll synthesis in plant cells, but its effect on mammalian cells is not clear. Previous work from our lab studying the non-target effects of metolachlor demonstrated low-level metolachlor exposure inhibits growth in human cell types such as fibroblasts, HepG2 liver cells and THP-1 alveolar macrophages and alters expression of several proteins involved in cell cycle control. But it is not well understood what other cellular processes are altered by exposure to metolachlor.
In the current study, DNA microarray analysis is used to screen for altered gene expression after exposure to increasing concentration of metolachlor. DNA microarray analysis requires isolation and purification of total RNA from HepG2 cells before and after treatment followed by hybridization with human cellular DNA. By using this method, we can determine which genes are being expressed at levels that differ from our untreated or control cells. Therefore, this technology allows for altered gene expression in treated vs. control cells.
After preliminary analysis, we expect to find differences in the expression of a family of genes involved in cell growth and cell cycle control as we have previously found differences looking at individual proteins in these processes between control and treated cells. We will generate heat maps of clustered genes and pathway analysis will be performed to determine affected pathways. This type of analysis has been demonstrated previously with other pesticides including atrazine and other chloroacetanilide family members.
Start Date
4-4-2017 1:00 PM
End Date
4-4-2017 4:30 PM
Faculty Advisor
Kavita Dhanwada
Department
Department of Biology
Copyright
©2017 Navinder Brar
Embargo Date
4-4-2017
Microarray Analysis of Gene Expression in Metolachlor Treated Human Liver Cells
Metolachlor, a commonly used pre-emergent pesticide in the Midwest, is a frequent contaminant of ground and surface waters. It inhibits protein and chlorophyll synthesis in plant cells, but its effect on mammalian cells is not clear. Previous work from our lab studying the non-target effects of metolachlor demonstrated low-level metolachlor exposure inhibits growth in human cell types such as fibroblasts, HepG2 liver cells and THP-1 alveolar macrophages and alters expression of several proteins involved in cell cycle control. But it is not well understood what other cellular processes are altered by exposure to metolachlor.
In the current study, DNA microarray analysis is used to screen for altered gene expression after exposure to increasing concentration of metolachlor. DNA microarray analysis requires isolation and purification of total RNA from HepG2 cells before and after treatment followed by hybridization with human cellular DNA. By using this method, we can determine which genes are being expressed at levels that differ from our untreated or control cells. Therefore, this technology allows for altered gene expression in treated vs. control cells.
After preliminary analysis, we expect to find differences in the expression of a family of genes involved in cell growth and cell cycle control as we have previously found differences looking at individual proteins in these processes between control and treated cells. We will generate heat maps of clustered genes and pathway analysis will be performed to determine affected pathways. This type of analysis has been demonstrated previously with other pesticides including atrazine and other chloroacetanilide family members.
Comments
Location: Maucker Union Oak Room