Fig. 8. Activation of PTEN is required for the TRPV4 inhibition induced growth suppression in colon cancer.

a Silencing of TRPV4 or HC-067047 induces dephosphorylation of PTEN. HCT-116 or SW620 cells were transfected with control siRNA (siCTL), TRPV4 siRNA#1 (siTRPV4#1) or TRPV4 siRNA#2 (siTRPV4#2) for 72 h, or treated with vehicle (0.1% DMSO) or HC-067047 (4 μΜ) for 72 h. The protein levels of phosphor-PTEN (Ser380/Thr382/383; p-PTEN), PTEN, and ACTB were analyzed by western bolt. b The effect of PTEN siRNA (siPTEN) on the inhibition of AKT-mTOR signaling, the decrease of cyclin D3 expression or the increase of apoptosis marker cleaved PARP and Caspase3 expression induced by TRPV4 silencing. HCT-116 cells were transfected with siCTL, siTRPV4#1 plus siCTL, siTRPV4#1 plus siPTEN for 72 h. c Silencing of TRPV4 or HC-067047 induces the nuclear localization of PTEN. HCT-116 or SW620 cells were transfected or treated as in (a). The immunofluorescent images were taken on a confocal microscope. Scale bar: 10 μm. d The effect of PTEN siRNA on the decrease of cell viability induced by TRPV4 silencing. Cell viability was assessed by MTT assay. e The effect of PTEN siRNA on the decrease of colony formation induced by TRPV4 silencing. All quantitative data shown represent the means ± SEM of at least three independent experiments. ^*P < 0.05 and ^#P < 0.001, versus the siTRPV4#1 plus siCTL group