Abstract
Objective: To evaluate the inhibition of proliferation of human osteosarcoma cells transfected with Pin1 anti-sense gene. Methods: Different doses of antisense Pin1 gene (0, 20, 50, 100, 200, 250 µL) were transfected into osteosarcoma MG-63 cells. The cells and culture supernatant before and after transfection were collected. The curve of cell growth was made by MTT method. The cell growth cycle and apoptosis were detected by FCM. The expression of Pin1 was detected by Western-blot and that of Pin1 mRNA by polymerase chain reaction (RT-PCR) respectively. Results: MTT and FCM assays indicated that the transfection by antisense Pin1 gene could inhibit MG-63 proliferation and induce apoptosis. Western-blot assays revealed that the antisense Pin1 gene-transfected MG-63 cells had weaker staining than those without transfected with antisense Pin1 gene, and staining intensity was negatively related with doses. The cells transfected by different doses of gene (0, 20, 50, 100, 200, 250 µL) had different absorbance rate: 0.854±0.136, 0.866±0.138, 0.732±0.154, 0.611±0.121, 0.547±0.109, 0.398±0.113, 0.320±0.151 respectively, with the difference being significant by F and q test (P<0.05). The expression of Pin1 mRNA had the similar results and its absorbance rate was 0.983±0.125, 0.988±0.127, 0.915±0.157, 0.786±0.125, 0.608±0.124, 0.433±0.130, 0.410±0.158 respectively (P <0.05). Conclusion: The expression of Pin1 mRNA in MG-63 cells could be inhibited by antisense Pin1 gene, so to reduce the expression of Pin1 and depress the proliferation of human osteosarcoma cells MG-63.
Similar content being viewed by others
References
Blum-Jensen P, Hunter T. Oncogenic kinase signalling. Nature, 2001, 411: 355–365.
Hanahan D, Weinberg RA. The hallmarks of cancer. Cell, 2000, 100: 57–70.
Lu KP, Liou YC, Zhou XZ. Pinning down the praline-directed phosphorylation signalling. Trends Cell Biol, 2002, 12: 164–172.
Ryo A, Liou YC, Lu KP, et al. Prolyl isomerase Pin1: a catalyst for oncogenesis and a potential therapeutic target in cancer. Cell Science, 2003, 116: 773–783.
Bao L, Kimzey A, Sauter G, et al. Prevalent overexpression of prolyl isomerase Pin1 in human cancers. Am J Pathol, 2004, 164: 1727–1737.
Riedel F. Expression of VEGF and inhibition of tumor angiogenesis by abrogation of VEGF in head and neck cancer. Laryngorhinootologie, 2003, 82: 436–437.
Whiteway A, Deru W, Prentice HG, et al. Construction of adeno-associated virus packaging plasmids and cells that directly select for AAV helper functions. J Virol Methods, 2003, 114: 1210.
Wulf GM, Liou YC, Ryo A, et al. Role of Pin1 in the regulation of p53 stability and p21 transactivation, and cell cycle checkpoints in response to DNA damage. Biol Chem, 2002, 277: 47976–47979.
Rippmann JF, Hobbie S, Daiber C, et al. Schnapp A: Phosphorylation-dependent proline isomerization catalyzed by pin1 is essential for tumor cell survival and entry into mitosis. Cell Growth Differ, 2000, 11: 409–416.
Fujimori F, Takahashi K, Uchida C, et al. Mice lacking Pin1 develop normally, but are defective in entering cell cycle from G0 arrest. Biochem Biophys Res Commun, 1999, 265: 658–663.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Xiong, W., Chen, A. & Guo, F. Inhibition of proliferation of human osteosarcoma cells transfected with PIN1 antisense gene. Chinese German J Clin Oncol 5, 294–297 (2006). https://doi.org/10.1007/s10330-005-0411-8
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10330-005-0411-8