Investigating Karyopherin B1: small molecule interactions for cancer therapy
type: CreativeWork
description: The advent of gene expression profiling studies has allowed for the identification of genes with potential as disease markers and therapeutic targets. Our laboratory identified the eukaryotic nuclear importer protein Karyopherin B1 (KpnB1), to be up-regulated in different cancer cell lines, including cervical and oesophageal as well as transformed cells. Inhibition of KpnB1 in these cells using small interfering RNA (siRNA) resulted in significant cancer cell death via apoptosis, suggesting KpnB1 is essential for cancer cell survival. Within our laboratory, we established that candidate small molecules targeted against KpnB1 identified using a rational drug design approach. The outcome of this research is for examine inhibitors of KpnB1 for potential as future anti-cancer agents using. Based on the long-term goal of this research, this particular project was aimed at investigating a small molecule inhibitor identified in our laboratory, known as Inhibitor of Nuclear Import-43 (INI-43) for its potential to bind to the nuclear importer, KpnB1. Using conventional assays as well as cutting edged techniques including circular dichrosim (CD) and isothermal titration calorimeter (ITC), an examination of INI-43 and its interactions with KpnB1 was made. In vitro analysis showed that INI-43 exhibits cytotoxic effects on cervical cancer cells with an IC₅₀ of ≈10μM and induces apoptotic cell death. The NFAT dual luciferase assay measured nuclear import of KpnB1 associated proteins, showing that INI-43 inhibits nuclear import/activity of NFAT in a dose dependent manner. Confocal microscopy of exogenous FRFP-KpnB1 as well as endogenous KpnB1 in the presence of INI-43 showed a change in the localisation of KpnB1 upon drug treatment. Both FRFP-KpnB1 and endogenous KpnB1 appear to be prevented from entering the nucleus, and is retained in the peri-nuclear space and the cytoplasm suggesting that INI-43 inhibits KpnB1 movement into the nucleus. To investigate KpnB1-INI-43 interactions…
abstract: The advent of gene expression profiling studies has allowed for the identification of genes with potential as disease markers and therapeutic targets. Our laboratory identified the eukaryotic nuclear importer protein Karyopherin B1 (KpnB1), to be up-regulated in different cancer cell lines, including cervical and oesophageal as well as transformed cells. Inhibition of KpnB1 in these cells using small interfering RNA (siRNA) resulted in significant cancer cell death via apoptosis, suggesting KpnB1 is essential for cancer cell survival. Within our laboratory, we established that candidate small molecules targeted against KpnB1 identified using a rational drug design approach. The outcome of this research is for examine inhibitors of KpnB1 for potential as future anti-cancer agents using. Based on the long-term goal of this research, this particular project was aimed at investigating a small molecule inhibitor identified in our laboratory, known as Inhibitor of Nuclear Import-43 (INI-43) for its potential to bind to the nuclear importer, KpnB1. Using conventional assays as well as cutting edged techniques including circular dichrosim (CD) and isothermal titration calorimeter (ITC), an examination of INI-43 and its interactions with KpnB1 was made. In vitro analysis showed that INI-43 exhibits cytotoxic effects on cervical cancer cells with an IC₅₀ of ≈10μM and induces apoptotic cell death. The NFAT dual luciferase assay measured nuclear import of KpnB1 associated proteins, showing that INI-43 inhibits nuclear import/activity of NFAT in a dose dependent manner. Confocal microscopy of exogenous FRFP-KpnB1 as well as endogenous KpnB1 in the presence of INI-43 showed a change in the localisation of KpnB1 upon drug treatment. Both FRFP-KpnB1 and endogenous KpnB1 appear to be prevented from entering the nucleus, and is retained in the peri-nuclear space and the cytoplasm suggesting that INI-43 inhibits KpnB1 movement into the nucleus. To investigate KpnB1-INI-43 interactions…
author: Strydom, Erin
language: eng
date published: 2016
date modified: 2026-07-22T22:23:46Z
provider: University of Cape Town
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Division of Medical Biochemistry

PropertyValue
property ID: Handle
value: http://hdl.handle.net/11427/22897