摘要
目的 :研究端粒酶RNA模板区特异性核酶对人低分化鼻咽癌CNE 2Z细胞增殖、凋亡的影响。方法 :用电转染的方法将带有绿色荧光蛋白GFP报道系统的端粒酶核酶基因的真核表达质粒PGFPuro teloRZ7 1及空载质粒PPAT GFP导入人低分化鼻咽癌CNE 2Z细胞 ;检测转染细胞的GFP表达情况、细胞增殖指数及凋亡。结果 :CNE 2ZGTR7 1细胞 (转染目的基因质粒PGFPuro teloRZ7 1的CNE 2Z细胞 )和CNE 2ZG细胞 (转染空载质粒PPAT GFP的CNE 2Z细胞 )有GFP表达 ,而未转染的CNE 2Z细胞无GFP表达。流式细胞仪检测显示 ,CNE 2ZGTR7 1细胞增殖指数 [(2 5 10 0±0 14 1) % ]明显低于CNE 2Z细胞 ,即未转染的细胞 [(5 3 663± 16 981) % ]和CNE 2ZG细胞 [(61 5 75± 5 166) % ] ,差异有统计学意义 ,P <0 0 1。CNE 2ZGTR7 1细胞传第 12代后有凋亡出现 ,CNE 2Z及CNE 2ZG细胞无凋亡。结论 :端粒酶核酶基因的导入使CNE 2Z细胞的增殖能力下降并可诱导CNE 2Z细胞凋亡。端粒酶RNA模板区可以作为鼻咽癌的一个治疗靶点 ,端粒酶RNA模板区特异性核酶可望成为有效的端粒酶抑制剂 。
OBJECTIVE:To study the effect of the proliferation and apoptosis of human low differentiated nasopharyngeal carcinoma (NPC) cell line CNE 2Z by a hammerhead ribozyme (telomerase ribozyme,teloRZ)directed against the RNA component of human telomerase (hTR) used as template.METHODS:A eukaryotic expression plasmid P GFPuro teloRZ7 1 containing teloRZ gene and the green fluorescent protein (GFP) reporter gene was constructed.Eukaryotic expression plasmid P GFPuro teloRZ7 1 and control plasmid P PAT GFP were transfected into human low differentiated NPC cell line CNE 2Z by electroporation, and then detected the expression of GFP,cellular proliferation and apoptosis.RESULTS:Green fluorescence were detected in CNE 2ZG cells transfected by control plasmid P PAT GFP and CNE 2ZGTR7 1 cells transfected by plasmid P GFPuro teloRZ7 1 . There was no GFP expression in CNE 2Z cells untransfected by any plasmid. The results of cell cycle analysis by FCM indicated that the cellular proliferation index (PI) of CNE 2ZGTR7 1 cells[(25 100±0 141)%]was significantly lower than those of CNE 2Z cells[(53 663±16 981)%] and CNE 2ZG cells[ (61 575±5 166)%], P <0 01.Apoptosis could be observed in CNE 2ZGTR7 1 cells which were transferred after 12 generation.There was no apoptosis occurring in CNE 2Z and CNE 2ZG cells.CONCLUSIONS:TeloRZ gene was electroporated successfully into CNE 2Z cells.TeloRZ can inhibit the proliferation and induce the apoptosis of CNE 2Z cells.The template region of telomerase RNA can be a target to treat NPC.TeloRZ may produce a marked effect of antisense nucleic acids technology for telomerase inhibition and cancer therapy.
出处
《肿瘤防治杂志》
2004年第1期38-42,共5页
China Journal of Cancer Prevention and Treatment
基金
广东省自然科学基金资助项目 (3 1963 )
