[Zhonghua yan ke za zhi] Chinese journal of ophthalmology · 2008
[RNA interference targeting aquaporin-1 on inhibition of human vascular endothelial cell migration in vitro].
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Zhang Hong-xu, Zhang Ming-chang, Chen Hong
Аннотация
OBJECTIVE: To investigate inhibition of human vascular endothelial cell migration in vitro by RNA interference targeting AQP-1, and the mechanism of corneal neovascularization.
METHODS: Experiment research. Human umbilical vein endothelial cell (HUVEC) was cultured, AQP-1 specific siRNA was designed and synthesized. siRNA was transfected into the HUVEC by Lipofectamine 2000. The mRNA of AQP-1 was assessed by reverse transcription-polymerase chain reaction (RT-PCR). Transwell migration assay and wound healing assay were used to detect the migration capability of HUVEC. The group comparison of AQP-1 mRNA expression was used one-way ANOVA test The result of transwell migration assay and wound healing assay was analysed with Independent-Sample T test.
RESULTS: The expression of AQP-1 mRNA was significantly (P < or = 0.05) suppressed after transfection. 24 h, 48 h and 72 h, after transfection, the expression of AQP-1 mRNA was 11.3%, 17.4% and 29.7% respectively compared to normal control (P = 0.004, 0.007, 0.002). Transwell migration assay and wound healing assay showed that the migration capability of HUVEC was decreased obviously (t = 10.813, P = 0.016 and t = 22.431, P = 0.027).
CONCLUSIONS: AQP-1 specific siRNA is effective in suppressing the AQP-1 expression and inhibiting the migration of the cultured HUVEC. Reducing the expression of AQP-1 can inhibit the corneal neovascularization.
METHODS: Experiment research. Human umbilical vein endothelial cell (HUVEC) was cultured, AQP-1 specific siRNA was designed and synthesized. siRNA was transfected into the HUVEC by Lipofectamine 2000. The mRNA of AQP-1 was assessed by reverse transcription-polymerase chain reaction (RT-PCR). Transwell migration assay and wound healing assay were used to detect the migration capability of HUVEC. The group comparison of AQP-1 mRNA expression was used one-way ANOVA test The result of transwell migration assay and wound healing assay was analysed with Independent-Sample T test.
RESULTS: The expression of AQP-1 mRNA was significantly (P < or = 0.05) suppressed after transfection. 24 h, 48 h and 72 h, after transfection, the expression of AQP-1 mRNA was 11.3%, 17.4% and 29.7% respectively compared to normal control (P = 0.004, 0.007, 0.002). Transwell migration assay and wound healing assay showed that the migration capability of HUVEC was decreased obviously (t = 10.813, P = 0.016 and t = 22.431, P = 0.027).
CONCLUSIONS: AQP-1 specific siRNA is effective in suppressing the AQP-1 expression and inhibiting the migration of the cultured HUVEC. Reducing the expression of AQP-1 can inhibit the corneal neovascularization.