Combining kallistatin gene therapy and meloxicam to treat hepatocellular carcinoma in mice

Abstract

Hepatocellular carcinoma (HCC) is one of the most common cancerrelated causes of death, and conventional treatments offer unsatisfactory response. We have previously reported that kallistatin gene therapy suppressed the growth of HCC tumors by its antiangiogenic activity, and meloxicam, a selective COX-2 inhibitor, inhibited proliferation and induced apoptosis of human HCC cells in vitro. The aim of this study was to determine whether combining kallistatin gene therapy and meloxicam could offer a better therapeutic effect to combat HCC in mice. A kallistatin expression plasmid was constructed and its expression was detected after intratumoral gene transfer. Both kallistatin gene therapy and meloxicam suppressed the growth of subcutaneous human HepG2 tumors established in BALB ⁄ c nude mice, and the combinational therapy showed a stronger effect in suppressing tumor growth, tumor angiogenesis and cell proliferation, and increasing cell apoptosis, than the respective monotherapies. Gene transfer of kallistatin inhibited tumor angiogenesis, and slightly inhibited cell proliferation and increased cell apoptosis in situ, but had no effect on expression of vascular endothelial growth factor, basic fibroblast growth factor, proliferating cell nuclear antigen, Bcl-2, Bax, or activation of caspase-3. Meloxicam therapy inhibited cell proliferation, induced cell apoptosis, reduced expression of proliferating cell nuclear antigen, increased activation of caspase-3, and upregulated Bax. Meloxicam also slightly inhibited tumor angiogenesis with no effect on the expression of vascular endothelial growth factor or basic fibroblast growth factor. Combining two novel anticancer agents, kallistatin targeting tumoral vascularization and meloxicam targeting cell proliferation and apoptosis, warrants investigation as a therapeutic strategy to combat HCC. (Cancer Sci 2009; 100: 2226-2233 H epatocellular carcinoma is the fifth most common cancer and the third leading cause of cancer-related mortality worldwide with an estimated incidence of over one million new cases each year.(1) Surgery offers a cure, but the rate of liver resection is <15% and recurrence rate remains as high as 50% after resection.(2) The conventional treatments including chemotherapy and radiotherapy offer unsatisfactory response. Therefore, new strategies to combat HCC are urgently needed.Solid tumors including HCC must establish an adequate vascular network to acquire necessary nutrition. Given that HCC is a hypervascular tumor, anti-angiogenic therapy offers a promising approach to treatment. Previous studies have revealed that angiogenesis inhibitors including TNP-470, (3) angiostatin, (4) endostatin, (5) soluble VEGF receptor 1,and vasostatin (6) are effective in treating HCC in experimental animal models. Kallistatin, a serine proteinase inhibitor, was first identified as a tissue kallikrein-binding protein,and has recently emerged as a novel inhibitor of angiogenesis. (8) Kallistatin inhibits the proliferation, migration, and adhesion of endothelial cells a...

Side effects of pain and analgesia in animal experimentation

“…Several studies have shown that NSAIDs, Coxibs and aspirin help to prevent cancer 56 . In particular, the effect of meloxicam has been described as inhibiting primary tumor growth in hepatocellular carcinoma or ovarian cancer mouse models 57,58 , as well as having an anti-metastatic effect in an orthotopic osteosarcoma mouse model 59 .…”

Kallistatin suppresses cancer development by multi-factorial actions

“…Kallistatin is an effective inhibitor of angiogenesis and inflammation. In murine models of tumor xenografts, kallistatin treatment inhibits the development of diverse cancers, including breast, liver, colon, lung and gastric carcinoma [29–34, 47]. Moreover, systemic delivery of the kallistatin gene dramatically inhibits experimental lung metastasis, inflammation and angiogenesis in tumor-bearing mice [34].…”

“…Kallistatin possesses anti-tumor activity as evidenced by inhibiting tumor growth and metastasis in animal models [29–34, 47]. For example, intramural injection of adenovirus carrying the human kallistatin gene into pre-established breast cancer xenografts in nude mice results in significant suppression of tumor growth and reduction of blood vessel numbers [29].…”

“…Furthermore, lentivirus-mediated kallistatin gene delivery by intravenous injection dramatically decreases tumor metastasis into lungs in association with reduced angiogenesis and inflammation, and enhances the survival of tumor-bearing mice [34]. In addition, a combination of local kallistatin gene therapy with meloxicam (a selective cyclooxygenase-2 inhibitor) was shown to have a better therapeutic effect in combating liver cancer in mice by inhibiting tumor growth and angiogenesis, and inducing apoptosis of human hepatocellular carcinoma cells [47]. Thus, addition of extrinsic kallistatin with various strategies by adenovirus, adeno-associated virus, lentivirus, plasmid DNA, or recombinant kallistatin protein, can retard tumor growth and progression in xenografted mice with breast, liver, stomach, colon and lung carcinomas.…”

Combined zoledronic acid and meloxicam reduced bone loss and tumour growth in an orthotopic mouse model of bone‐invasive oral squamous cell carcinoma

“…41 While there are no published reports of the effects of meloxicam on clinical or preclinical in vivo OSCC studies, Meloxicam has been shown to induce tumor cell apoptosis in human patients with esophageal cancer 42 and reduce tumor growth in animal models of other forms of cancer including osteosarcoma, 43 hepatocellular carcinoma 44 and ovarian carcinoma. 45 …”