ABSTRACT
Objective To investigate the effect of anti-sense osteopontin(ANOPN)on proliferation and metastasis of esophagus cancer cells.Methods An OPN gene recombinant expression vector plasmid was constructed by RT-PCR from human umbilical vein endothelial cell gene and cloned into a mammalian expression vector pcDNA3.1(+).PcDNA3.1-ANOPN was introduced by LipofectinTM.Positive cell clones(ECA-ANOPN),vector-transfected cells ECA-vect and blank cell ECA were used as three groups.RT-PCR and immunocytochemistry assay were used to investigate the expressions of OPN mRNA and protein.The metastasis characteristics of cells were studied by Transwell method.Results The vector was constructed successfully,the sequencing result was identical with that reported in GenBank.Compared with vector-transfected cells(ECA-vect cells) and ECA cells,the growth rate of ECA-ANOPN cells was significantly slowed(P
ABSTRACT
To evaluate the inhibitory effects of fas gene transduction on oesophageal cancer cells in vitro, the eukaryotic expression vector fas pBK was constructed and transfected into oesophageal carcinoma cells EC109. Western blot results showed high expression of Fas protein in gene transfected cells Fas EC109. Cell growth curve and plating efficiency test revealed that Fas EC109 had longer population doubling time and lower plating efficiency compared with control cells. Results of MTT assay showed increased sensitivity of Fas EC109 cells to CDDP, VCR and 5 FU. All these data suggested that transgenic expression of fas gene could effectively inhibit the proliferation and increase the drug sensitivity of oesophageal carcinoma cells in vitro.