Abstract 3617: Inhibition of KLF4 expression in resistant B-NHL cell lines inhibited cell growth and sensitized the cells to drug-induced apoptosis

Abstract

Abstract Kruppel-like factor 4 (KLF4) is a member of the KLF4 zinc-finger containing transcription factor family. Reported studies have indicated the involvement of KLF4 in the regulation of proliferation, apoptosis, and differentiation of B-cells and B-cell malignancies. We have recently reported that KLF4 is overexpressed in pediatric Burkitt lymphomas and is a predictive biomarker for survival. In addition, overexpression of KLF4 predicted unresponsiveness to CHOP treatment (Valencia-Hipolito A et al Leuk Lymphoma. 2014;55:1806-14). Preliminary findings demonstrated that the transcription factor Yin Yang 1 (YY1) plays, in part, a role in the regulation of KLF4 expression in B-NHL. We hypothesized that (1) the chemical inhibition of KLF4 may result in the inhibition of proliferation, induction of apoptosis, and sensitization to drug-induced apoptosis in B-NHL cell lines and (2) the inhibition of YY1 would mimic the chemical inhibition of KLF4. Analysis of two B-NHL cell lines revealed that the expression of KLF4 was high in Ramos and low Raji as compared to normal B-cells. Treatment with increasing concentrations (0.5-10 μM) of Kenpaullone (a KLF4 inhibitor) induced in Ramos cells both the inhibition of proliferation and cell survival and the induction of apoptosis; however, there was no effect on the treatment of Raji cells. Treatment of Ramos cells with the combination of Kenpaullone and CDDP potentiated apoptosis as compared to treatment with either the chemical inhibitor or CDDP used alone. The finding with the chemical inhibitor was validated with the transfection of Ramos cells with siRNA-KLF4. In addition, treatment with Kenpaullone inhibited the transcriptional expression of KLF4 (using a reporter assay system with pKLF4-GFP). Additional studies demonstrated that the transfection of Ramos cells with siRNA-YY1 resulted in significant inhibition of KLF4 expression and correlated with the inhibition of proliferation and the induction of apoptosis. The latter is due, in part, as KLF4 suppresses the extrinsic apoptotic pathway by inhibiting the activation and cleavage of caspases 7, 9, and 3.. Our previous analysis in TMA of pediatric lymphomas demonstrated, in all of the tumor tissues, the presence of a positive correlation between the expression of KLF4 and YY1 and that this correlation was markedly significant in the Burkitt subtype. In conclusion, the overexpression of KLF4 may be responsible, in part, in the pathogenesis, malignancy, and drug resistance of B-NHL lymphomas. In addition, the present findings suggest that the chemical inhibition of KLF4 by Kenpaullone treatment or the inhibition of YY1 may be considered as targets for therapeutic intervention in the treatment of B-lymphoma overexpressing KLF4, when used alone or in combination with sub-toxic chemo/immune-drugs. Current studies are evaluating the role of KLF4 inhibition in vivo using B-NHL tumor xenografts models. Citation Format: Mayra R. Montecillo-Aguado, Gabriel G. Vega, Hector Mayani, Sara Huerta-Yepez, Rogelio Hernández-Pando, Otoniel Martinez-Maza, Benjamin Bonavida, Mario I. Vega. Inhibition of KLF4 expression in resistant B-NHL cell lines inhibited cell growth and sensitized the cells to drug-induced apoptosis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3617. doi:10.1158/1538-7445.AM2015-3617