OBJECTIVE: To study the role of aquaporin 5(AQP5) in regulating the cell proliferation and apoptosis of human colorectal cancer cells and explore the possible mechanism. METHODS: A small interfering RNA (siRNA) targeting AQP5 was used to suppress endogenous AQP5 expression in the human colorectal cancer cell lines COLO 205 and SW480, and the transfection efficiency of AQP5 siRNA was determined using immunofluorescence assay and PCR. The changes in the proliferation of the transfected cells was evaluated with MTT assay, and the cell apoptosis was analyzed using Annexin V-FITC/PI and TUNEL assays; the changes of Bax and Bcl2 expressions in the cells were determined using RTPCR and Western blotting. RESULTS: Transfection with AQP-5-siRNA resulted in a significant reduction (up to 90%) of AQP-5 expression in COLO 205 and SW480 cells. MTT assay showed that AQP-5-siRNA transfection significantly inhibited the cell proliferation compared NS siRNA transfection (P<0.05). Flow cytometric analysis revealed significantly increased apoptotic rate of cells following AQP-5-siRNA transfection compared with NS?siRNA transfection(P<0.05). Real-time quantitative RT-PCR and Western blotting demonstrated that AQP-5-siRNA transfection significantly increased Bax and Bcl-2 expressions at both mRNA and protein levels in the cells. CONCLUSION: AQP5-siRNA can promote apoptosis of colorectal cancer cells in vitro possibly in relation to its effects on Bax and Bcl expressions.
OBJECTIVE: To study the role of aquaporin 5(AQP5) in regulating the cell proliferation and apoptosis of humancolorectal cancer cells and explore the possible mechanism. METHODS: A small interfering RNA (siRNA) targeting AQP5 was used to suppress endogenous AQP5 expression in the humancolorectal cancer cell lines COLO 205 and SW480, and the transfection efficiency of AQP5 siRNA was determined using immunofluorescence assay and PCR. The changes in the proliferation of the transfected cells was evaluated with MTT assay, and the cell apoptosis was analyzed using Annexin V-FITC/PI and TUNEL assays; the changes of Bax and Bcl2 expressions in the cells were determined using RTPCR and Western blotting. RESULTS: Transfection with AQP-5-siRNA resulted in a significant reduction (up to 90%) of AQP-5 expression in COLO 205 and SW480 cells. MTT assay showed that AQP-5-siRNA transfection significantly inhibited the cell proliferation compared NS siRNA transfection (P<0.05). Flow cytometric analysis revealed significantly increased apoptotic rate of cells following AQP-5-siRNA transfection compared with NS?siRNA transfection(P<0.05). Real-time quantitative RT-PCR and Western blotting demonstrated that AQP-5-siRNA transfection significantly increased Bax and Bcl-2 expressions at both mRNA and protein levels in the cells. CONCLUSION:AQP5-siRNA can promote apoptosis of colorectal cancer cells in vitro possibly in relation to its effects on Bax and Bcl expressions.
Authors: Lindsey A Torre; Freddie Bray; Rebecca L Siegel; Jacques Ferlay; Joannie Lortet-Tieulent; Ahmedin Jemal Journal: CA Cancer J Clin Date: 2015-02-04 Impact factor: 508.702