整合素连接激酶对人视网膜色素上皮细胞增殖和迁移的影响

目的:探讨RNA干扰抑制整合素连接激酶（integrin-linked kinase, ILK）对人视网膜色素上皮（human retinal pigment epithelium,hRPE）细胞增殖和迁移的影响。方法:体外培养hRPE细胞,设计并合成特异性人ILK的RNA干扰片段,阳离子脂质体转染入人视网膜色素上皮细胞,利用RT-PCR及Western blot半定量检测siRNA对ILK基因及蛋白表达的抑制作用,MTT方法检测转染前后siRNA对细胞增殖的抑制作用。损伤愈合实验和Transwell实验观察人视网膜色素上皮细胞迁移能力的改变。结果:培养的hRPE细胞存在ILK的基因转录表达,siRNA显著抑制hRPE细胞ILK的mRNA和蛋白表达（P<0.01）。MTT、损伤愈合实验和Transwell实验提示转染ILK-siRNA后人视网膜色素上皮细胞的增殖、迁移能力有明显下降,与正常对照组相比有统计学差异（P<0.05）。结论:特异性ILK-siRNA能有效抑制ILK在hRPE的mRNA和蛋白的表达并显著降低hRPE的增殖和迁移能力。

Effect of integrin-linked kinase on the proliferation and migration of human retinal pigment epithelium cells

AIM:To investigate the effect of RNA interference targeting integrin-linked kinase (ILK) on the proliferation and migration of cultured human retinal pigment epithelium (hRPE)cells.METHODS:hRPE cells were cultured, and their morphology and staining features were determined. ILK specific siRNA was designed and synthesized. siRNA was transfected into hRPE cells with Lipofectamine 2000. The effects of transfection were observed using a fluorescence microscope. RT-PCR and Western blot were used to semiquantitatively evaluate the mRNA and protein expressions of ILK. MTT assay was used to detect the proliferation of hRPE cells. Wound healing assay and transwell migration assay were used to detect the migration capability of hRPE cells. RESULTS:ILK was found in normal culture hRPE cells. The mRNA and protein expressions of ILK significantly decreased by siRNA transfection(P<0.01).The proliferation of hRPE cells also significantly decreased(P<0.05). The migration capability of hRPE significantly decreased by siRNA transfection, with statistically significant difference compared with the normal control group(P<0.05).CONCLUSION:ILK-siRNA transfection significantly decreases the mRNA and protein expressions of ILK, as well as the proliferation and migration abilities of hRPE cell’s.