Activation of Phosphatidylinositol 3-Kinase Is Sufficient for Cell Cycle Entry and Promotes Cellular Changes Characteristic of Oncogenic Transformation

ABSTRACT

Using a new inducible form of phosphatidylinositol 3-kinase (PI 3-kinase) we have found that PI 3-kinase activation has the following effects on cell growth and proliferation. (i) Activation of PI 3-kinase was sufficient to promote entry into S phase of the cell cycle within several hours. This was shown by activation of cyclin-dependent kinase 4 (Cdk4) and Cdk2 and by the induction of DNA synthesis. (ii) PI 3-kinase activation alone was not, however, sufficient to provide for progression through the entire cell cycle. Instead, prolonged activation of PI 3-kinase in the absence of serum stimulation resulted in apoptosis. It is possible that the cells undergo apoptosis because the PI 3-kinase-induced entry into the cell cycle is abnormal. For example, we found that the cyclin E-Cdk2 complex, which normally disappears after entry into S phase of the cell cycle, fails to be downregulated following induction by PI 3-kinase. (iii) Finally, we found that prolonged activation of PI 3-kinase in the presence of serum resulted in cellular changes that resemble those associated with oncogenic transformation. The cells reached high densities, were irregular and refractile in appearance, and formed colonies in soft agar. In contrast, neither PI 3-kinase nor serum stimulation alone could induce these changes. Our results suggest that activation of PI 3-kinase promotes anchorage-independent cell growth and entry into the cell cycle but does not abrogate the growth factor requirement for cell proliferation.

ACKNOWLEDGMENTS

We thank Catherine Tribouley for generously providing the Myc · ER expression vector. We thank Laurie Goda for the speedy synthesis of oligonucleotides and Moijgan Amir-Ebrahimi and Jeff Tucker for DNA sequence analysis. We are grateful to Kang Dai, Bert Pronk, Christoph Reinhard, Kelly Smith, Anne Roulston, and Ning Lee for sharing their expertise on cell cycle regulation and apoptosis. We thank Kang Dai, Bert Pronk, Christoph Reinhard, Steve Harrison, A. B. Jefferson, Nicholas Marini, and especially Kelly Smith, Kevin Ramer, and Lisa Molz for many helpful comments on the manuscript.