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Tetracycline-inducible protein expression in pancreatic cancer cells: Effects of CapG overexpression

World Journal of Gastroenterology 2011年第15期17卷 1947-1960页

作者： Sarah Tonack Sabina Patel Mehdi Jalali Taoufik Nedjadi Rosalind E Jenkins Christopher Goldring John Neoptolemos eithne costello Liverpool CR-UK Centre Department of Molecular and Therapeutic Cancer MedicineUniversity of Liverpool Liverpool L69 3GA United Kingdom MRC Centre forDrug Safety Science Department of Pharmacology and Therapeutics School of Biomedical Sciences University of Liverpool L69 3GA United Kingdom National Institute for Health Research Pancreatic Biomedical Research Unit Liverpool L69 3GA United Kingdom

AIM: To establish stable tetracycline-inducible pancreatic cancer cell lines. METHODS: Suit-2, MiaPaca-2, and Panc-1 cells were transfected with a second generation reverse tetracycline-controlled transactivator protein (rtTA2S-M2), under the control of either a cytomegalovirus (CMV) or a chicken β-actin promoter, and the resulting clones were ***: Use of the chicken (β-actin) promoter proved superior for both the production and maintenance of doxycycline-inducible cell lines. The system proved versatile, enabling transient inducible expression of a variety of genes, including GST-P, CYP2E1, S100A6, and the actin capping protein, CapG. To determine the physiological utility of this system in pancreatic cancer cells, stable inducible CapG expressors were established. Overexpressed CapG was localised to the cytoplasm and the nuclear membrane, but was not observed in the nucleus. High CapG levels were associated with enhanced motility, but not with changes to the cell cycle, or cellular proliferation. In CapG-overexpressing cells, the levels and phosphorylation status of other actin-moduating proteins (Cofilin and Ezrin/Radixin) were not altered. However, preliminary analyses suggest that the levels of other cellular proteins, such as ornithine aminotransferase and enolase, are altered upon CapG induction. CONCLUSION: We have generated pancreatic-cancer derived cell lines in which gene expression is fully controllable.

关键词： Pancreatic cancer cells Tetracycline-inducible CapG Suit-2 Panc-1 MiaPaCa-2

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