Increased Expression Levels Of Igfbp-5 And Gpx1 In Bxpc-3 Pancreatic Cancer Cell Lines.

No Thumbnail Available

Links to Files

Author/Creator ORCID

Date

2017

Type of Work

Department

Biology

Program

Master of Science

Citation of Original Publication

Rights

This item is made available by Morgan State University for personal, educational, and research purposes in accordance with Title 17 of the U.S. Copyright Law. Other uses may require permission from the copyright owner.

Abstract

Pancreatic cancer is a highly fatal disease with death usually occurring within one year of diagnosis. American Cancer Society predicts that by the end of 2017, there will be 53,670 new cases and 43,090 fatalities of pancreatic cancer. Despite the aggressiveness of this disease, little is known about its mechanism of development. It has been speculated that reactive oxygen species (ROS) damages cellular DNA causing deleterious mutations which leads to altered proteins and cellular functions including transformation of normal to malignant phenotype. Insulin-like growth factor binding protein-5 (IGFBP-5) is associated with regulation of biological functions such as cell growth, differentiation, apoptosis, cell adhesion, and metastasis: processes that are known to be critical in the etiology of cancer. Additionally, Glutathione peroxidase 1 (GPx1) is an anti-oxidant enzyme that detoxifies hydrogen peroxide in cells thereby minimizing oxidative stress. We therefore, hypothesized that expression of IGFBP-5 and GPx1 may be altered in pancreatic cancer cells. To address our hypothesis, we analyzed the expression of IGFBP-5 and GPx1, in the pancreatic cancer cell line, BxPC-3 relative to the non-malignant pancreatic cell line, HPDE-6. Both cell lines were cultured in 5% CO2 incubators at 37oC and nuclear extracts were subsequently prepared. The protein concentrations were determined by BCA assay and the proteins resolved by SDS polyacrylamide gel electrophoresis (SDS-PAGE). The protein bands were stained with Coomassie Blue and de-stained with water. Antibodies against IGFBP-5, and GPx1 were used to assess protein expression via Western Blot Analysis. Interestingly, our results showed that multiple proteins with molecular weights ranging from 10 kDa to 300 kDa including IGFBP-5 and GPx1 were differentially expressed in BxPC-3 compared to the normal cell line. This expression was further verified by Western Blot Analysis. These findings are consistent with our hypothesis and implicate the involvement of IGFBP-5 and GPx1 in the etiology of pancreatic cancer. Further characterization of these proteins may yield useful insights essential in the understanding of the mechanisms underlying pancreatic cancer development. Furthermore, this research may be critical in identifying biomarkers useful in therapeutic intervention of pancreatic cancer.