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Transduction of Human Cells with Polymer-complexed Ecotropic Lentivirus for Enhanced Biosafety
Published on: July 24, 2011
The FUS1 gene inhibits EC109 cell growth mediated by a lentivirus vector
1School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, PR China. zhengff2004@gmail.com
Abstract:
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.
Insights
The FUS1 gene may suppress tumors in esophageal carcinoma. Introducing FUS1 into EC109 cells inhibited their growth, suggesting its potential as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Esophageal carcinoma is a significant global health concern.
- Identifying novel therapeutic targets is crucial for improving patient outcomes.
- The role of the FUS1 gene in esophageal carcinoma remains largely unexplored.
Purpose of the Study:
- To investigate the functional role of the FUS1 gene in esophageal carcinoma.
- To determine if FUS1 expression influences the proliferation of EC109 cells.
- To assess the potential of FUS1 as a tumor suppressor gene in esophageal cancer.
Main Methods:
- Messenger RNA (mRNA) expression of FUS1 was quantified using reverse transcription polymerase chain reaction (RT-PCR).
- FUS1 gene was amplified, cloned into a lentiviral vector, and transfected into EC109 cells.
- Cell proliferation was assessed using MTT assays and plate colony formation assays.
Main Results:
- FUS1 expression was lower in EC109 cells compared to control cell lines.
- Transfection of FUS1 into EC109 cells resulted in over 80% efficiency within 48 hours.
- Cell growth was inhibited by approximately 40% in FUS1-transfected cells, confirmed by both MTT and colony formation assays.
Conclusions:
- The FUS1 gene exhibits reduced expression in esophageal carcinoma cells.
- FUS1 gene introduction significantly inhibits the proliferation of esophageal carcinoma cells in vitro.
- FUS1 may function as a tumor suppressor gene, warranting further in vivo investigation for therapeutic potential.
