[Gene marking of human colon cancer LoVo cells with enhanced green fluorescent protein].
OBJECTIVE
The aim of this study is to obtain a human colon cancer cell line marked with enhanced green fluorescent protein (EGFP) gene.


METHODS
The G1FP retroviral vector, carrying EGFP gene, was constructed. When combined with liposome-mediated transfection with cross infection, EGFP virus producer cells were generated. The human colon cancer LoVo cells were transduced with EGFP retrovirus. The integration and expression of EGFP gene were analyzed with polymerase chain reaction (PCR) and flow cytometry.


RESULTS
The ecotropic EGFP virus was obtained by cocultivation of packaging cells for two weeks, while the amphotropic virus producer cells Am12/G1FP, expressing high-level EGFP, were generated through cross infection with a titer of 1.1 x 10(5) infectious particles/ml. The genetically modified LoVo/GFP cells gave off strikingly bright green fluorescence, in which the integration of EGFP provirus was confirmed by PCR analysis.


CONCLUSIONS
The LoVo/GFP cells, which express high-level green fluorescence, may provide an useful tool for the study of micrometastasis of colon cancer in vivo.
Chen
S
S
Department of General Surgery, First Hospital, Suzhou University, Suzhou 215006, China.
Fu
J
J
Zhang
Z
Z
D000818:Animals; D003001:Cloning, Molecular; D003110:Colonic Neoplasms; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics*; D051379:Mice; D008819:Mice, Nude; D012190:Retroviridae / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins