Inhibition of the aquaporin 3 water channel increases the sensitivity of prostate cancer cells to cryotherapy.
Aquaporins (AQPs) are intrinsic membrane proteins that facilitate selective water and small solute movement across the plasma membrane. In this study, we investigate the role of inhibiting AQPs in sensitising prostate cancer cells to cryotherapy. PC-3 and DU145 prostate cancer cells were cooled to 0, -5 and -10 degrees C. The expression of AQP3 in response to freezing was determined using real-time quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. Aquaporins were inhibited using mercuric chloride (HgCl(2)) and small interfering RNA (siRNA) duplex, and cell survival was assessed using a colorimetric assay. There was a significant increase in AQP3 expression in response to freezing. Cells treated with AQP3 siRNA were more sensitive to cryoinjury compared with control cells (P<0.001). Inhibition of the AQPs by HgCl(2) also increased the sensitivity of both cell lines to cryoinjury and there was a complete loss of cell viability at -10 degrees C (P<0.01). In conclusion, we have shown that AQP3 is involved directly in cryoinjury. Inhibition of AQP3 increases the sensitivity of prostate cancer cells to freezing. This strategy may be exploited in the clinic to improve the efficacy of prostate cryotherapy.
Ismail
M
M
Department of Oncology, Postgraduate Medical School, University of Surrey, Guildford GU2 7WG, UK. ms18273@gmail.com
Bokaee
S
S
Morgan
Richard
R
Davies
J
J
Harrington
K J
KJ
Pandha
H
H
D051400:Aquaporin 3 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D017679:Cryotherapy; D005455:Fluorescent Antibody Technique; D006801:Humans; D008297:Male; D008627:Mercuric Chloride / Q000494:pharmacology*; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
C496027:AQP3 protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051400:Aquaporin 3; D008627:Mercuric Chloride