CpG Immunomer DNA Enhances Antisense Protein Kinase A RIα Inhibition of Multidrug-Resistant Colon Carcinoma Growth in Nude Mice: Molecular Basis for Combinatorial Therapy

Abstract

CpG DNAs induce cytokines, activate natural killer cells, and elicit vigorous T-cell response leading to antitumor effects. Antisense oligodeoxynucleotides targeted against the RIalpha subunit of protein kinase A (antisense PKA RIalpha) induce growth arrest, apoptosis, and differentiation in a variety of cancer cell lines in vitro and in vivo. This study investigated the use of a combinatorial therapy consisting of the RNA-DNA second-generation antisense PKA RIalpha and the CpG immunomer (CpG DNA linked through 3'-3' linkage containing two accessible 5' ends). HCT-15 multidrug-resistant colon carcinoma growth in nude mice was used as an experimental model. The inhibitory effect on tumor growth and apoptotic activity of antisense RIalpha and CpG immunomer, singly and in combination, were measured by tumor growth, levels of RIalpha subunit, and antiapoptotic and proapoptotic proteins. Effect on host-immune system was measured by mouse spleen size, interleukin-6 (IL-6) levels in mouse blood, and nuclear factor-kappaB (NF-kappaB) transcription activity in mouse spleen cells. In combination, CpG immunomer and antisense PKA RIalpha induced additive/supra-additive effect on the inhibition of tumor growth. Antisense RIalpha but not CpG immunomer increased Bax and Bak proapoptotic protein levels and decreased Bcl-2 and RIalpha protein levels in tumor cells. CpG immunomer but not antisense RIalpha induced an enlargement of mouse spleen, increased IL-6 levels in mouse blood, and increased NF-kappaB transcription activity in mouse spleen cells. These results show that type I PKA down-regulation and induction of apoptosis in tumor cells by antisense PKA RIalpha, and host-immune stimulation by CpG immunomer are responsible at the molecular level for the supra-additive effects of tumor growth inhibition. Thus, antisense PKA RIalpha and CpG immunomer in combination work cooperatively and as tumor-targeted therapeutics to treat human cancer.