端粒酶hTRT基因反义寡核苷酸逆转前列腺癌细胞恶性表型的研究  

Malignant Phenotypes Reverse Effects on Prostate Cancer Cells by Antisense Oligonucleotide Targeting Telomerase hTRT Gene

作　　者：童强松[1] 郑丽端[2] 叶哲伟[1] 曾甫清[1] 鲁功成[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院泌尿外科,武汉430022 [2]华中科技大学同济医学院附属协和医院病理科,武汉430022

出　　处：《华中科技大学学报（医学版）》2003年第5期487-490,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：卫生部科研基金资助项目 (No 98 1 136 )

摘　　要：目的　探讨反义技术阻断人类端粒酶催化亚单位 (hTRT)基因表达对前列腺癌细胞生物学行为的影响。方法　设计、合成异硫氰酸荧光素标记的hTRT基因反义寡核苷酸 (ASODN) ,在脂质体介导下转染前列腺癌细胞系PC3 M ,运用荧光显微镜观察ASODN在癌细胞内的分布 ,MTT比色分析、流式细胞仪、克隆形成实验检测癌细胞体外生长活性 ,RT PCR和PCR ELISA法检测hTRT基因表达及端粒酶活性。结果　 0 5～ 2 0 μmol/LASODN转染 12～ 36h后 ,ASODN逐渐聚集于细胞核 ,PC3 M细胞增殖活性抑制 34 1%～ 79 6 % (P <0 0 1) ,细胞周期阻滞于G0 /G1期 ,克隆形成能力抑制 6 1 2 6 % (P <0 0 1) ,hTRTmRNA水平降低 2 4 4 8%～ 86 73% (P <0 0 1) ,端粒酶活性抑制 2 4 74 %～ 76 72 % (P <0 0 1)。结论　运用ASODN阻断hTRT基因表达 ,降低端粒酶活性 。Objective To explore the effects of blocking expression of human telomerase reverse transcriptase (hTRT) on biological behavior of prostate cancer cells by antisense technique. Methods The 5′ isothiocyananate (5′ FITC) conjugated antisense oligonucleotide (ASODN) for hTRT was designed and synthesized to transfect prostate cancer cell line PC3 M with liposome. The cellular distribution of ASODN was observed by fluorescence. The in vitro cellular growth activities were detected by MTT colormetric, flow cytometric and clone formation assays. The hTRT expression and telomerase activities were respectively detected by RT PCR and PCR ELISA methods. Results When being transfected with 0 5-2 0 μmol/L ASODN for 12-36 h, ASODN gradually localized within nucleus. The growth of cancer cells were inhibited by 34 1 %-79 6 % ( P <0 01). The cell cycle was arrested at G 0/G 1 phase, with the clone formation abilities being inhibited by 61 26 % ( P <0 01). The hTRT mRNA level was decreased by 24 48 %-86 73 % ( P <0 01), while the telomerase activities being reduced by 24 74 %-76 72 % ( P <0 01).Conclusion The malignant phenotypes of prostate cancer could be reversed by reducing the hTRT gene expression and telomerase activities with ASODN.

关 键 词：端粒酶 HTRT基因 反义寡核苷酸 前列腺癌 细胞恶性表型 研究 

分 类 号：R737.25[医药卫生—肿瘤]