hTERT基因反义核酸对Raji细胞端粒酶活性的影响  

Effects of hTERT antisense oligodeoxynucleotides on telomerase activity in Raji cells

作　　者：李文瑜[1] 张洹[2] 何冬梅[2] 

机构地区：[1]广东省人民医院肿瘤中心,广东广州510080 [2]暨南大学医学院血液病研究所,广东广州510632

出　　处：《中国病理生理杂志》2004年第2期196-199,共4页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金重点项目 (0 2 1195 ) ;广州市科技计划重点基金资助目 (2 0 0 1-Z - 0 37- 0 1)

摘　　要：目的 :观察人端粒酶逆转录酶 (hTERT)基因的全硫代修饰反义寡核苷酸 (ASODN)对Raji细胞端粒酶活性的影响。方法 :端粒酶聚合酶链反应 -酶联免疫测定法检测Raji细胞的端粒酶活性 ;采用逆转录 -聚合酶链反应方法检测hTERT基因mRNA的表达水平 ;以间接免疫荧光标记法通过流式细胞仪检测hTERT蛋白表达含量的变化。结果 :hTERTASODN作用于Raji细胞后 ,hTERTmRNA表达水平明显降低 ,ASODN与Raji细胞共同孵育 4 8h ,hTERT蛋白表达阳性率降为 5 0 15 %± 3 4 9% ,72hhTERT蛋白表达阳性率降为 33 35 %± 2 75 %。而hTERT正义核酸作用 2 4、4 8、72h对hTERT蛋白表达阳性率 (6 5 % )无显著影响 (P >0 0 5 )。hTERT基因ASODN作用于Raji细胞 4 8h ,其端粒酶活性明显降低 ,作用 72h ,端粒酶活性明显受到抑制 ,分别与正义寡核苷酸组、空白对照组相比有显著差异 (P <0 0 5 )。结论 :hTERT基因反义寡核苷酸特异性地抑制Raji细胞的端粒酶活性。AIM: To explore the effect of hTERT gene antisense phosphorothioate oligodeoxynucleotide (ASODN) on telomerase activity in Raji cells. METHODS: Polymerase chain reaction enzyme-linked immuonassay (PCR-ELISA) was used to determine telomerase activity. The expression levels of hTERT mRNA and protein were assayed by reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence using fluoresce isothiocyanate (FITC) label,respectively. RESULTS: RT-PCR and immunofluorescence assay showed that the expression levels of hTERT mRNA and protein from Raji cells decreased with time after hTERT ASODN treatment. There was no difference in hTERT mRNA and protein levels between control and SODN-treated cells. Telomerase activity decreased when Raji cells were treated with ASODN for 48 h. Telomerase activity of Raji cells was significantly inhibited when treated with ASODN for 72 h. There was no difference in telomerase activity levels between control and hTERT SODN-treated cells. CONCLUSION: hTERT ASODN could inhibit telomerase activity in Raji cells.

关 键 词：基因 端粒酶逆转录酶 寡核苷酸类 反义 RAJI细胞 端粒酶 

分 类 号：R733.7[医药卫生—肿瘤]