Phospholipase C gamma 1 regulates the Rap GEF1-Rap1 signalling axis in the control of human prostate carcinoma cell adhesion.
Phospholipase Cgamma1 (PLCgamma1) is activated downstream of a variety of extracellular stimuli and has previously been implicated in the regulation of motility responses central to tumour cell invasion. In this study, we used a novel RNAi vector system to achieve conditional PLCgamma1 knockdown in PC3LN3 human prostate carcinoma cells for further evaluation of PLCgamma1 in tumour cell biology. Using this approach, we revealed a role for PLCgamma1 in the regulation of PC3LN3 cell adhesion that appears to be independent of its effects on tumour cell chemotactic migration and spreading in response to extracellular matrix. Subsequent microarray analysis of PLCgamma1-knockdown cells revealed Rap GEF1 mRNA to be decreased in response to PLCgamma1 loss. This translated into a decrease in Rap GEF1 protein levels and a significant loss of Rap1 activity in PLCgamma1-knockdown cells. Transient knockdown of Rap GEF1 caused a reduction in PC3LN3 adhesion while overexpression of Rap GEF1 rescued the PLCgamma1 knockdown-induced adhesion defect. These data highlight control of the Rap GEF1-Rap1 molecular switch as a specific requirement for PLCgamma1-mediated tumour cell adhesion.
Cancer Research UK Centre for Cancer Therapeutics, The Institute of Cancer Research, Sutton, UK.
J C
JC
Peak
N P
NP
Jones
S
S
Hobbs
M
M
Katan
S A
SA
Eccles
D002277:Carcinoma / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D002448:Cell Adhesion / Q000235:genetics; D045744:Cell Line, Tumor; D002633:Chemotaxis / Q000235:genetics; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008297:Male; D009419:Nerve Tissue Proteins / Q000378:metabolism / Q000502:physiology*; D051966:Phospholipase C gamma / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology; D015398:Signal Transduction / Q000502:physiology*; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism / Q000502:physiology*
D020662:Guanine Nucleotide Exchange Factors; D009419:Nerve Tissue Proteins; C403525:RAPGEF2 protein, human; D051966:Phospholipase C gamma; D020827:rap1 GTP-Binding Proteins