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| The Effect of URI Gene Stable Expression on Cell Proliferation in SMMC-7721 Cell Line |
| JI Hui, WANG Qi, ZHONG Lei, WEI Wen-xiang |
| Dept of Cell Biology, School of Biology and Basic Medicine Science, Medical College, Soochow University, Suzhou 215123, China |
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Abstract Objective: To establish the cell lines which can stably express the RPB5-mediating protein (URI) and study the effects of URI gene on the biological function of human hepatoma SMMC-7721 cells. Methods: Get PFLAG-CMV-4-URI from the E.coli which was transfected the plasmid. Then to identify the recombinant eukaryotic expression by enzyme analysis, RT-PCR Transient transfected the plasmid PFLAG-CMV-4-URI into SMMC-7721 cells by Lipofectamine reagent. G418 selected the cell clone which stably expressed URI. The URI expression were detected by RT-PCR and cell proliferation was measured by MTT assay. Plate colony forming assay was used to observe the clone formation ability of cell lines. Results: The positive clones which stably expressed URI were selected. Result of RT-PCR and enzyme analysis showed that URI gene could stably expression in SMMC-7721 cells. MTT assay showed that URI promoted the growth of SMMC-7721 cells. And the URI gene could also improve the formation rate of cell clones. Conclusion: URI gene can improve the proliferation of SMMC-7721 cells probably by helping the cell through the G2/M checkpoint.
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Received: 10 December 2012
Published: 25 June 2013
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