端粒酶反義寡核苷酸抑制視網膜色素上皮細胞增殖的實驗研究

目的使用脂質體介導的端粒酶反義寡核苷酸封閉端粒酶基因在視網膜色素上皮細胞的表達從而抑制視網膜色素上皮細胞的增殖,爲增殖性玻璃體視網膜病燮的治療探索一條基因治療途徑.方法使用端粒重復序列擴增法(Telomericrepeat amplification protocol,TRAP)定性、定量檢測濃度爲0umol/L、2umol/L、4umol/L、6umol/L、8umol/L脂質體介導的端粒酶反義寡核苷酸及濃度爲4umol/L的正義寡核苷酸在視網膜色素上皮細胞端粒酶活性表達.結果TRAP結果顯示随着端粒酶反義寡核苷酸濃度的增高,視網膜色素上皮細胞端粒酶活性逐漸降低,8umol/L幾乎没有表達,正義寡核苷酸很少抑制視網膜色素上皮細胞端粒酶活性.結論脂質體介導的端粒酶反義寡核苷酸能抑制視網膜色素上皮細胞端粒酶的活性從而抑制視網膜色素上皮細胞的增殖.

Suppression ofTelomerase Antisense Oligodeoxynucleotide on Growth of Human RPE Cells

Objective To inhibit human retinal pigment epithelial (hRPE) cell proliferation by blocking telomerase gene expression with liposome-mediated telomerase antisense oligodeoxynucleotide(AS-ODN), in order to search for a new approach of genetic therapy for proliferative vitreoretinopathy (PVR). Methods The liposome-mediated synthetic phosphorothioate telomerase AS-ODN labeled with FAM fluorescence in 5'ends were delivered to RPE cells at different concentrations of 0 uM, 2 uM, 4 uM, 6 uM and 8 uM; then telomerase activity was examined by telomerase repeat amplification protocol (TR AP) and quantitative analysis. Results Telomerases AS-ODN at 4 uM, 6 uM and 8 uM significantly inhibited hRPE cell proliferation. TRAP showed the expression of liposome-mediated telomerase AS-ODN in hRPE cells became weaker with the increase of concertrations and there was almost no expression at 8um. Conclusion The liposome-mediated telomerase AS-ODN at some concentrations can sequence-specifically supress telomerase expression in hRPE cells and cellular proliferative activity. These findings establish a rationale for investigating the potential use of telomerase AS-ODN as a novel therapeutical tool for treatment of PVR.