PCR-ELISA定量检测as-hTERT对HepG2.2.15细胞端粒酶活性的抑制作用  

Detection of telomerase activity in as-hTERT-treated HepG2.2.15 cells by PCR-ELISA in vitro

作　　者：刘素侠[1] 孙汶生[1] 马春红[1] 韩丽辉[1] 宋静[1] 

机构地区：[1]山东大学医学院免疫学研究所

出　　处：《山东大学学报（医学版）》2002年第4期292-293,共2页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助课题 ( 30 0 70 34 1)

摘　　要：目的 :采用PCR ELISA方法探讨人类端粒酶催化亚单位 (hTERT)基因关键区段的反义寡核苷酸 (as hTERT)对HepG2 .2 .1 5细胞端粒酶活性的抑制作用。方法 :通过PCR合成as hTERT及无关对照序列 ,体外作用于HBVDNA转染的HepG2 .2 .1 5细胞 ,用PCR ELISA定量检测反义核酸作用后的端粒酶活性 ,MTT法观察as hTERT对细胞生长的抑制作用 ,ELISA法检测as hTERT对HBsAg和HBeAg表达的影响。结果 :as hTERT作用浓度为 1 0 μmol/L时 ,定量检测端粒酶活性 ,A450 nm值为 0 .42 ,低于无关序列与生理盐水对照的A450 nm值 (分别为 1 .46、1 .49) ,as hTERT可抑制细胞生长 ,对HBsAg和HBeAg的最佳抑制率分别为 76%和5 6%。结论 :as hTERT在体外可明显降低HepG2 .2 .1 5细胞的端粒酶活性 ,抑制细胞的生长 ,并可影响HBV抗原表达。Objective: To detect the telomerase activity and anti HBV effect in HepG2.2.15 cells treated with as hTERT in vitro. Methods: The sequences of antisense phosphorothioate oligodeoxynucleotides complementary to the initiator of hTERT(as hTERT) and random control were synthesized through PCR. HepG2.2.15 cells were given 10 μmol/L as hTERT.Telomerase activity was detected by PCR ELISA method and the anti HBV effect was assayed with ELISA. Results: Growth arrest was observed in cells given as hTERT and inhibition of telomerase activity was detected in as hTERT treated cells. The best inhibition rates of HBsAg and HBeAg were 76% and 56% respectively. Conclusion: As hTERT has anti tumour activity and may be used as tumour cell inhibitors.

关 键 词：抑制作用 人类端粒酶催化亚单位 反义寡核苷酸类 聚合酶链反应-酶联免疫分析 

分 类 号：R730.3[医药卫生—肿瘤]