Ki-67-directed antisense therapy in an orthotopic renal cell carcinoma model.

Abstract

The Ki-67 antigen is only present in proliferating cells. We have shown previously that phosphorothioate-modified antisense oligonucleotides (ON) against this antigen are potent antitumoral agents in bladder and prostate cancer-derived cells. Since ON are known to accumulate in vivo in the kidney, high local effectivity may be expected. Here, we evaluated and characterized antitumoral effects in an orthotopic renal cell cancer (RENCA) model. RENCA cells were incubated with antisense and control ON in the presence of a cationic lipid. Uptake studies were performed with FITC-labeled ON. Ki-67 protein analysis after ON treatment was performed by immunohistochemical staining. For animal studies, 1 x 10(5) RENCA cells were implanted under the renal capsule of Balb/c mice. Antisense and control ON were injected intraperitoneally daily for 14 days. Tumor weights and status of metastasis were documented after sacrifice. Furthermore, vessel density in tumor tissues was determined by CD31 immunolabeling. Antisense treatment of RENCA cells resulted in specific reduction of the Ki-67 protein and inhibition of cell growth. A substantial cellular uptake of labeled ON was noted in vitro and in vivo. The growth of orthotopically implantated syngeneic kidney tumors in immunocompetent mice was significantly inhibited in antisense-treated animals (p < 0.05). Furthermore, lung metastases were noted in 10% of antisense-treated animals compared to 30-40% in control groups. Immunohistochemical staining of the vessel density showed no significant difference among treatment groups. The results demonstrate that Ki-67-directed antisense oligonucleotides are potent inhibitors of target protein expression and proliferation of tumor cells in vitro, and of tumor growth and lung metastasis formation in murine renal cell carcinoma whereas tumor vascularization is not significantly affected.