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  • Title: [Down-regulation of VEGF expression and up-regulation of PEDF expression in human corneal epithelial cells under hypoxic condition by RNA interference of HIF-1α].
    Author: Zhang X, Chen J, Yao M, Liu X, Zhou Q.
    Journal: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi; 2018 Oct; 34(10):880-884. PubMed ID: 30554580.
    Abstract:
    Objective To investigate the effect of short hairpin RNA (shRNA) targeting hypoxia-inducible factor-1α (HIF-1α) on the expression of vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) in spontaneously immortalized human corneal epithelial cells (S-ihCECs) under hypoxic condition. Methods Three kinds of shRNAs targeting the sites of HIF-1α mRNA were designed and synthesized, and then transfected into S-ihCECs in vitro. After transfection, the cells were exposed to 1 mmol/L CoCl2 for 6 hours. HIF-1α, VEGF and PEDF mRNA expressions were detected by real-time quantitative PCR. HIF-1α and VEGF protein levels were analyzed by Western blotting. PEDF level was tested by ELISA. Results After the shRNA interference, the expressions of HIF-1α and VEGF mRNA decreased by (70.9±2.8)% and (51.9±15.7)%, respectively when compared with the hypoxia group. The expressions of HIF-1α and VEGF proteins were both down-regulated. The expression of PEDF mRNA increased (1.22±0.18) fold and the protein was up-regulated. Conclusion ShRNA targeting HIF-1α can effectively silence the expression of HIF-1α, reduce the expression of VEGF and up-regulate the expression of PEDF.
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