[Effect of transferred NK4 gene on biological characteristics of human pancreatic cancer cell line SW1990].
OBJECTIVE
Hepatocyte growth factor (HGF) plays an important role in the regulation of migration, invasion,and angiogenesis of cancer via the activation of its receptor, c-Met. NK4 is not only an antagonist of HGF but also an angiogenesis inhibitor. The blockade of HGF/c-Met signal pathway and tumor angiogenesis may be a new strategy for cancer treatment. This study was designed to construct eukaryotic expressing vector of NK4 gene, transfer it into human pancreatic cancer cell line SW1990, and observe the effect of transfected NK4 gene on the biological behaviors of SW1990 cells,and its expression in SW1990 cells.


METHODS
The recombinant of pcDNA3/hNK4 plasmid was digested by restrictive enzyme,NK4 gene was cloned into a high effective eukaryotic expressing vector pRC/CMV2, and the recombinant of pRC/CMV2-hNK4 plasmid was transiently introduced into SW1990 cells by lipofectamine. Reverse transcriptase-polymerase chain reaction (RT-PCR),and Western blot were used to detect the expression of NK4 at mRNA, and protein levels,respectively. Migration, and invasion capabilities of the transfected cells were evaluated by Transwall chamber, and Matrigel invasion chamber, respectively.


RESULTS
Expressions of NK4 gene after lipofectamine mediated transfection were observed in SW1990 cells, expected fragment of 453 bp has been amplified by RT-PCR, and Western blot analysis showed positive expression of NK4 protein (50 KDa). NK4 gene had no inhibitory effect on the growth of SW1990 cells (2.2x10(5) vs 2.5x10(5), P >0.05), while it had significantly suppressive effect on the migration and invasion of SW1990 cells driven by HGF or fibroblasts (P< 0.01).


CONCLUSIONS
NK4 gene transfection may inhibit spreading and invasion of pancreatic cancer cells, which would play an important role in the anti-metastasis therapy for pancreatic cancer.
Department of Gastroenterology,The Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai,Guangdong,519000, P.R.China. lairenxu@hotmail.com
Ren-Xu
RX
Lai
Shi-Zhen
SZ
Yuan
Toshikazu
T
Nakamura
D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D018076:DNA, Complementary / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D017228:Hepatocyte Growth Factor / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008934:Mitogens / Q000235:genetics* / Q000502:physiology; D009361:Neoplasm Invasiveness; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D010957:Plasmids; D019859:Proto-Oncogene Proteins c-met / Q000235:genetics* / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
D018076:DNA, Complementary; C066855:HGF protein, human; D008934:Mitogens; D011994:Recombinant Proteins; D017228:Hepatocyte Growth Factor; D019859:Proto-Oncogene Proteins c-met