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目的 研究反义寡核苷酸能否封闭原癌基因c-myc的表达并抑制培养的人结膜成纤维细胞的增生,为青光眼滤过术后滤过泡的瘢痕化探索新的治疗途径。方法 取人结膜下筋膜进行成纤维细胞的培养,脂质体Lipofectamine~(TM)2000的介导下分别将正义c-myc、反义c-myc转入细胞中,MTT法检测其对细胞增生活性的影响,RT-PCR扩增检测其对细胞mRNA表达水平的影响。结果 反义c-myc能显著抑制培养的人结膜成纤维细胞的增生,并抑制其c-myc mRNA的表达,而正义c-myc无此作用。反义c-myc的最适浓度为2μM。结论 反义c-myc通过阻断人结膜成纤维细胞中原癌基因c-myc表达来抑制细胞的增生。
Objective To investigate whether antisense oligonucleotides can block the expression of proto-oncogene c-myc and inhibit the proliferation of cultured human conjunctival fibroblasts, and explore a new therapeutic approach for the scarring of filtration bleb after glaucoma filtration. Methods Human subconjunctival fascia was taken for culture of fibroblasts. Lipofectamine 2000 was used to transfect c-myc and antisense c-myc into cells, respectively. Proliferative activity, RT-PCR amplification to detect the impact of cell mRNA expression levels. Results Antisense c-myc significantly inhibited the proliferation of cultured human conjunctival fibroblasts and inhibited the expression of c-myc mRNA, whereas c-myc did not. The optimal concentration of antisense c-myc was 2 μM. Conclusion Antisense c-myc inhibits cell proliferation by blocking c-myc expression in human conjunctival fibroblasts.