hTERT反义寡核苷酸对瘢痕疙瘩成纤维细胞增殖及端粒酶活性的影响  

Inhibition of hTERT antisense oligodeoxynucleotide on proliferation and telomerase activity in keloid fibroblasts

作　　者：黄勇[1] 林立新[1] 王鹏[1] 王学明[1] 王玉婷[1] 姜蕾[1] 

机构地区：[1]青岛大学医学院附属烟台毓璜顶医院烧伤整形科,山东省264000

出　　处：《中华临床医师杂志（电子版）》2012年第18期38-41,共4页Chinese Journal of Clinicians(Electronic Edition)

基　　金：烟台市科技发展计划(2011112)

摘　　要：目的研究hTERT基因反义寡核苷酸(ASODN)对瘢痕疙瘩成纤维细胞生长增殖及端粒酶活性的抑制作用,探讨其在瘢痕疙瘩治疗中的作用。方法应用ASODN封闭瘢痕疙瘩成纤维细胞hTERT基因的表达,不同浓度不同时间以ASODN作用于体外培养的瘢痕疙瘩成纤维细胞,细胞计数和MTT法检测成纤维细胞生长增殖情况,端粒酶PCR-ELISA法检测不同处理浓度和时间成纤维细胞端粒酶活性。结果 ASODN作用于瘢痕疙瘩成纤维细胞72h后,成纤维细胞生长受抑,增殖能力减弱,并具有浓度、时间依赖性。端粒酶活性检测显示:ASODN0.5、1.0和1.5μmol/L作用组OD450-690值分别为0.612±0.038、0.535±0.027、0.423±0.023,与空白对照组(0.898±0.058)比较,差异有统计学意义(P<0.05);正义寡核苷酸1.0μmol/L作用组(OD450-690值为0.893±0.042)与空白对照组比较,差异无统计学意义(P>0.05)。结论 hTERT基因ASODN能抑制瘢痕疙瘩成纤维细胞的生长增殖,降低成纤维细胞端粒酶活性是其重要作用机制之一。通过抑制端粒酶活性进行抗瘢痕疙瘩治疗可能是一个新途径。Objective To investigate the inhibition of hTERT antisense oligodeoxynucleotide(ASODN)on the proliferation and telomerase activity in keloid fibroblasts and to explore its anti-keloid effect.Methods Keloid fibroblasts in culture medium in vitro were given different concentrations of hTERT ASODN and were observed in different time.After treated by hTERT ASODN the proliferation of the fibroblasts was measured by cell count and MTT assay method,the changes of telomerase activity of keloid fibroblasts treated with hTERT ASODN was determined with PCR-ELISA method.The data was analyzed by statistical software(SPSS 11.5).Results The results showed that after sealing hTERT gene with ASODN for 72 hours,the fibroblasts growth was repressed and the ability of proliferation decreased,and the effect was specific in sequence and dependent in dose and time.OD450-690 values were 0.898±0.058,0.612±0.038,0.535±0.027,0.423±0.023 respectively,as the fibroblasts were treated with 0,0.5,1.0,1.5 μmol/L ASODN for 72 hours.The difference was significant as compared 0.5,1.0,1.5 μmol/L groups with 0 μmol/L ASODN group respectively(P<0.05),but the difference was no significant when compared 1.0 μmol/L SODN group(0.893±0.042)with untreated group(0.898±0.058)(P>0.05).Conclusions hTERT-ASODN can suppress growth and proliferation of keloid fibroblasts.Decreasing the telomerase activity of keloid fibroblasts may be one of the most important mechanisms.That hTERT-ASODN inhibited telomerase activity in keloid fibroblasts is an important pathway that may play a key role in the anti-keloid therapy.

关 键 词：瘢痕疙瘩 端粒 末端转移酶 寡核苷酸类 反义 

分 类 号：R622[医药卫生—整形外科]