E-cadherin mediates aggregation-dependent survival of prostate and mammary epithelial cells through the retinoblastoma cell cycle control pathway.
E-cadherin and the retinoblastoma tumor suppressor (Rb) are traditionally associated with diverse regulatory aspects of cell growth and differentiation. However, we have discovered new evidence, which suggests that these proteins are functionally linked in a physiologic pathway required for cell survival and programmed cell death. Pharmacological activation of protein kinase C (PKC) or inducible overexpression and activation of the alpha isozyme of PKC (PKCalpha) resulted in approximately 60% apoptosis of mammary and prostate epithelial cells. Interestingly, the surviving cells had undergone dramatic aggregation concurrent with increased E-cadherin expression. When aggregation was inhibited by the addition of an E-cadherin-blocking antibody, apoptosis increased synergistically. We hypothesized that survival of the aggregated population was associated with contact-inhibited growth and that apoptosis might result from aberrant growth regulatory signals in non-aggregated, cycling cells. This hypothesis was confirmed by experiments that demonstrated that E-cadherin-dependent aggregation resulted in Rb-mediated G1 arrest and survival. Immunoblot analysis and flow cytometry revealed that hypophosphorylated Rb was present in non-aggregated, S phase cultures concurrent with synergistic cell death. We have also determined that the loss of membrane E-cadherin and subsequent hypophosphorylation of Rb in luminal epithelial cells preceded apoptosis induced by castration. These findings provide compelling evidence that suggests that E-cadherin-mediated aggregation results in Rb activation and G1 arrest that is critical for survival of prostate and mammary epithelial cells. These data also indicate that Rb can initiate a fatal growth signal conflict in non-aggregated, cycling cells when the protein is hypophosphorylated as these epithelial cells enter S phase.
Day
M L
ML
Department of Surgery, Section of Urology, University of Michigan and the University of Michigan Comprehensive Cancer Center, Ann Arbor, Michigan 48109, USA. mday@umich.edu
Zhao
X
X
Vallorosi
C J
CJ
Putzi
M
M
Powell
C T
CT
Lin
C
C
Day
K C
KC
D017209:Apoptosis; D001940:Breast / Q000166:cytology*; D015820:Cadherins / Q000378:metabolism*; D002449:Cell Aggregation; D002453:Cell Cycle / Q000502:physiology*; D002460:Cell Line; D002470:Cell Survival; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D016193:G1 Phase; D006801:Humans; D008297:Male; D011467:Prostate / Q000166:cytology*; D011493:Protein Kinase C / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000502:physiology*
D015820:Cadherins; D016160:Retinoblastoma Protein; D011493:Protein Kinase C