Marked regression of liver metastasis by combined therapy of ultrasound‐mediated NFkB‐decoy transfer and transportal injection of paclitaxel, in mouse

Abstract

Nuclear factor-kappaB (NFkB) plays a pivotal role in cancer progression. In this study, we developed a decoy cis-element oligodeoxyribonucleic acid against NFkB-binding site (NFkB-decoy), which effectively inhibits NFkB activity, and tested the effect of combined therapy comprising local transfection of NFkB-decoy into the liver and transportal injection of paclitaxel on cancer growth and metastasis using an orthotopic murine model of colon cancer liver metastasis. For NFkB-decoy transfection, we employed a novel approach using ultrasound exposure with an echocardiographic contrast agent, Optison. We examined the influence of NFkB-decoy transfer on susceptibility to paclitaxel in cancer cells and the mechanism involved using several in vitro analysis systems. We then studied the in vivo effect of combined NFkB-decoy transfer and paclitaxel in preventing cancer progression using a murine model of liver metastasis created by splenic injection of a human colon cancer cell line, HT29. In vitro experiments, including MTT-assay, fluorescence-activated cell sorter and cDNA array analysis, revealed that NFkB-decoy transfer significantly increased the susceptibility of cancer cells to paclitaxel, and that decreased expression of anti-apoptotic genes along with increased expression of genes relevant to the apoptosis-promotor may be involved. In vivo experiments showed that local transfection of NFkB-decoy into the liver followed by portal injection of paclitaxel effectively induced cancer cell apoptosis in the liver metastasis, and significantly prolonged animal survival compared to controls, without notable side effects. In conclusion, a combination of local NFkB-decoy transfer into the liver and transportal injection of paclitaxel may be a safe and effective new therapy for liver metastasis. ' 2007 Wiley-Liss, Inc.Key words: apoptosis; decoy cis-elements ODN; portal-injection; colon cancer; HT29 Nuclear factor-kappaB (NFkB) is one of the most critical transcription factors for regulating the gene expression of various antiapoptotic proteins, such as Bcl-XL (Bcl2 like 1), TNF receptorassociated factor (TRAF)-1, c-IAP and p21 as well as inflammatory cytokines such as IL-1, IL-8 and interferon-g. 1-3 It is generally assumed that NFkB-induced factors promote apoptotic resistance, transformation, cell growth, metastasis and angiogenesis in neoplastic tumors. 4 Basal NFkB activity is often increased in various types of cancer by several activated oncogenes such as ras and raf. 5 The survival of some cancer cells, such as hematological malignancies and prostate cancer, clearly depends on NFkB. [6][7][8] Therefore, inhibition of NFkB activity is an attractive approach to cancer therapy, and indeed has been attempted for many types of cancer. 9 Unfortunately, the results have not always been promising. Inhibition of NFkB alone does not dramatically induce cell death in most solid cancers, perhaps because many types of tumor can upregulate NFkB-independent anti-apoptotic pathways. Combined therapy involving inhibition of NFkB act...

Transfection of NF-κB decoy oligodeoxynucleotide suppresses pulmonary metastasis by murine osteosarcoma

“…40,41 The intravenous injection of 'naked' NF-kB decoy ODN in an animal model showed accumulation in the kidney (30% of the dose per g tissue), but little accumulation in other organs. 42 Therefore, alternative delivery methods such as local administration, 43,44 combination use with cationic liposome as a carrier, 40 and physical stimulation by ultrasound wave 45,46 have been reported. In other series of study, we directly injected 'naked' NF-kB decoy ODN into a LM8 tumor mass and examined the biodistribution and the inhibitory effects of the decoy in a murine spontaneous pulmonary metastasis model.…”

Sonoporation Delivery of Interleukin-27 Gene Therapy Efficiently Reduces Prostate Tumor Cell Growth In Vivo

“…Other studies have used sonoporation-based strategies to deliver immunomodulatory cytokine gene therapy encoding IFNc (Sakakima et al, 2005) or IL-12 (Suzuki et al, 2010), with significant tumor size reductions in mouse models of hepatocellular and ovarian carcinoma, respectively. Liver metastasis regression was accomplished by combined therapy of an NFjB oligodeoxynucleotide decoy delivered by sonoporation and transportal injection of paclitaxel (Azuma et al, 2008). Based on these studies and our present report, we suggest that sonoporation gene transfer might offer significant potential as a cancer therapy, and the effects of cytokines or other therapies might be augmented if used in combination with chemotherapy-based approaches.…”

The expression of SIRT1 regulates the metastaticplasticity of chondrosarcoma cells by inducing epithelial-mesenchymal transition

“…SW1353 cells (5 × 10 5 /300 μl/mouse) were slowly injected in 8-week-old nude mice via the splenic vein through a 27-gauge needle and the spleen was then removed. The mice were sacrificed after 6 weeks, the livers were harvested and liver metastasis nodule were measured18.…”