Differential expression of Nuclear Factor kappa B and anticancer properties of NBD peptide in human urothelial carcinoma

Abstract

Bladder cancer BC is a recurrent and often chemoresistant cancer that ranks second among genitourinary tract cancers Nuclear Factor kappaB is a critical transcription factor capable of driving disease progression upon constitutive activation in BC Nemo Binding Domain peptide NBD is a part of NFkappaB pathway with antiinflammatory activity This study delineates the differential expression of NFkappaB heterodimers and the anticancer properties of NBD peptide in BC Cytoscape was used for insilico NFkappaB heterodimer analysis and NBD peptide was validated by SVM algorithm based applications Immunohistochemistry was used for NFkappaB heterodimer expression and NBD peptide anticancer activity was analyzed invitro Cytoscape results revealed overexpression of RelA and suppression of p50 precursor in BC without hindering target protein expression Immunohistochemical localization showed nuclear RelAp50 in low grade and only RelA expression in high grade Cytoplasmic RelAp50 expression remained extensive in all high and low grade tissues RelA expression was proportional to the disease progression whereas the differential expression of nuclear p50 in regions of squamous differentiation and muscle invasion indicated its diagnostic potential NBD peptide cytotoxicity was assessed on T24 BC cells and noncancerous cells like NIH3T3L1 CHO Vero Selective dose dependent cytotoxicity was exhibited by NBD peptide on T24 cells while being nontoxic to noncancerous cells Combination chemotherapy with NBD and standard drugs was tested on T24 cells T24 cell viability after 48hours of NBD peptide treatment was 50 percent at 18 dot 5 micromoles and it increased to about 78 percent when treated with a combination of NBD with standard drugs T24 cells accumulated in subG0G1 phase as they underwent necrosis and lost the ability to migrate when treated with 100 micromoles NBD Thus NBD peptide exhibited selective cytotoxicity on T24 cells and the combination with standard drugs demonstrated excellent inhibition of BC newline