Inducible Activation of NFkappaB in Lour Grade and High Grade Transitional Carcinoma Cell Lines

Shin, Myung Sik; Rackley, Raymond; Appell, Rodney

Abstract

PURPOSE: While the exact inflammatory and immunological mechanisms involved in the induction of immunotherapy for superficial bladder carcinoma are mostly unknown, many of these mechanisms are mediated through the activation of transcription factors involved in signal transduction pathways Nuclear Factor Kappa B(NFkappaB) signal transduction pathways induce transcription factors that activate genes encoding immunological proteins such as cytokines and cell Surface maskers. This study investigates the activation pattern of NFkappaB by lipopolysaccharide(LPS), interferon gamma(INFgamma), and tissue necrosis factor alpha(TNFalpha) in low(RT4) and high(T24) grade transitional carcinoma cell lines.
MATERIALS AND METHODS
Low and high grade transitional carcinoma cell lines were cultured with Eagle's minimum essential medium. Using electrophoretic mobility shift assays(EMSA) of whole cell extracts from cell cultures of RT4 and T24 after exposure to LPS, INFgamma and TNFalpha, activation of NFkappaB complex has been demonstrated. Degradation of IkappaB has been demonstrated by Western blot analysis using IkappaBalpha /MAD-3.
RESULTS
NFkappaB complex was induced by TNFalpha in both RT4 and T24 cells as determined by EMSA. NFalphaB complex was also Induced by INFgamma in the RT4 cells but not in the T24 cells. However in LPS treatment, the NFkappaB complex was strongly induced in T24 cells and the induction was very weak in RT4 cells. Inhibitor of NFkappaB (IkappaBalpha) was degraded rapidly after LPS treatment in the T24 cells as determined by Western blot analysis with IkappaBalpha specific antibody. However, the level of IkappaBalpha protein was same in the RT4 cells before and after LPS treatment.
CONCLUSIONS
Identification of differences in low and high grade tumor cell lines in the inducibility of transcription factors by LPS and INFgamma provide an opportunity for understanding the observed differences in the mechanisms within the signal transduction pathways will ultimately create disease specific as well as patient specific treatments for these malignant urothelial disorders.