**6. Hydodynamics-based NK4 gene therapy for colon cancer inhibition**

Hydrodynamic delivery has emerged as the simplest and effective method for intracellular delivery of subjective genes in rodents; this process requires no special equipment. The system employs a physical force generated by the rapid injection of a large volume of solution into a blood vessel to enhance the permeability of endothelium and the plasma membrane of the parenchyma cells, such as hepatocytes, to facilitate a delivery of the substance into cells (Bonamassa *et al*., 2011). Using this technique in mice, we established an endocrine delivery system for NK4 that leads to an inhibition of the malignant behavior of cancers, as follows.

Prostate carcinoma r-NK4, sc Inhibition of growth, Davies G *et al*., (PC-3 cells, (osmotic pump) Anti-angiogenesis Int J Cancer 106: sc, Mouse) 348-354 (2003)

Ovarian carcinoma NK4 gene, Inhibition of metastasis, Saga Y *et al*., (HRA cells, Stable Prolonged survival Gene Ther 8: ip, Mouse) transfection 1450-1455 (2001)

Lymphoma Adeno-NK4, Inhibition of growth, Kikuchi T *et al*., (E.G7-OVA cells, intra-tumor Anti-angiogenesis, Blood 100: sc, Mouse) (with DC) Induction of CTL 3950-3959

Multiple myeloma Adeno-NK4, im Inhibition of growth, Du W *et al*., (KMS11/34 cells, Anti-angiogenesis, Blood 109: sc, Mouse) Enhanced apoptosis 3042-3049

Melanoma Adeno-NK4, iv Inhibitions of growth Kishi Y *et al*., (B16F10 cells, and metastasis, Cancer Sci 100: sc, Mouse) Anti-angiogenesis 1351-1358

Glioblastoma r-NK4, Inhibition of growth, Brockmann MA (U-87 MG cells, intra-tumor Anti-angiogenesis, *et al*., Clin Cancer

Breast carcinoma r-NK4, sc Inhibition of growth, Martin TA *et al.,* (MDAMB231 cells, Anti-angiogenesis Carcinogenesis sc, Mouse) 24: 1317-1323 (2003)

Intra-brain, Mouse) Enhanced apoptosis Res 9:

Adeno-NK4, adenoviral vector carrying NK4 cDNA; r-NK4, recombinant NK4 protein; sc, subcutaneous; iv, intravenous; ip, intraperitoneal; im, intramuscular; DC, dendritic cells; and CTL,

Table 1. Representative studies to show therapeutic effects of NK4 on distinct types of

**6. Hydodynamics-based NK4 gene therapy for colon cancer inhibition** 

Hydrodynamic delivery has emerged as the simplest and effective method for intracellular delivery of subjective genes in rodents; this process requires no special equipment. The system employs a physical force generated by the rapid injection of a large volume of solution into a blood vessel to enhance the permeability of endothelium and the plasma membrane of the parenchyma cells, such as hepatocytes, to facilitate a delivery of the substance into cells (Bonamassa *et al*., 2011). Using this technique in mice, we established an endocrine delivery system for NK4 that leads to an inhibition of the malignant behavior of

(2003)

(2007)

(2009)

 4578-4585 (2003)

C. Reproductive system:

D. Hematopoietic system**:** 

E. Other organ or tissues**:**

cytotoxic T lymphocytes.

cancers, as follows.

tumors in animal models
