Objective The effect of FUS1 gene on esophageal carcinoma cell line, EC109, was investigated.
Methods The mRNA expression level of FUS1 gene was detected by RT-PCR technique in the esophageal carcinoma cell lines of SHEE, SHEEC and EC109. The full length of FUS1 gene was amplified by method of PCR technique from the total RNA of the umbilical mesenchymal stem cells. FUS1 gene was cloned into pSL6-IRES-EGFP vector and identified with PCR, digestion and sequencing. The recombinant plasmid of pSL6-FUS1-IRES-EGFP was packaged in 293T cells and the lentivirus was collected. The infection effect was observed by transfection into EC109 cells. The transfected cells growth were determined by means of MTT.
Results: The expression of FUS1 gene in EC109 cells was weaker than that of SHEE and SHEEC cells. The lentivirus vector of pSL6-FUS1-IRES-EGFP was obtained identified by means of PCR, digestion, sequencing. The lentivirus vector was packaged in 293T cells and collected. EC109 cells were infected by the lentivirus. The transfection efficiency was more than 80% observed after 48h infection. The cell growth was inhibited by about 37% compared with the control group.
Conclusion: The recombinant lentivirus vector of FUS1 was successfully cloned and the preparation lentivirus may high effectively transfect cells. Over-expression of FUS1 gene could suppress the growth of esophageal carcinoma cells. The results suggested that FUS1 gene might be a candidate tumor suppressor gene for the gene therapy of esophageal carcinoma, which need be further confirmed by the results in vivo.
98th AACR Annual Meeting-- Apr 14-18, 2007; Los Angeles, CA