转染
分子生物学
生物
慢病毒
细胞培养
基因
基因表达
细胞生长
病毒学
遗传学
病毒
病毒性疾病
作者
B Zhang,Xiao Xu,Zhongtian Qi,Peng Lu,Bin Qiu,Xiaoxi Huo
标识
DOI:10.1080/09674845.2013.11669925
摘要
The effects of the FUS1 gene on the oesophageal carcinoma cell line EC109 are investigated. The messenger RNA (mRNA) expression level of the FUS1 gene was detected by a reverse transcription polymerase chain reaction (RT-PCR) technique in the cell lines SHEE, SHEEC and EC109. The full length of the FUS1 gene was amplified using a PCR technique from the total RNA of umbilical mesenchymal stem cells. The FUS1 gene was cloned into a pSL6-IRES-EGFP vector and identified by PCR, digestion and sequencing. The recombinant pSL6-FUS1-IRES-EGFP plasmid was transfected into 293FT cells and the resulting lentivirus was collected. The growth of EC109 cells after transfection with lentivirus containing the FUS1 gene was determined by MTT assay and plate colony formation. Expression of the FUS1 gene in EC109 cells was weaker than that in SHEE, SHEEC cells and human umbilical vein endothelial cells (HUVEE; used as a control). Transfection efficiency was more than 80% after 48 h. Cell growth assessed by MTT assay was inhibited by about 40% compared with the control group; a finding that was in accordance with the plate colony formation results. The results suggest that the FUS1 gene might be a candidate tumour suppressor gene for the treatment of oesophageal carcinoma; however, these results require confirmation in in vivo studies.
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