CAJ | 학술논문

目的：探讨载体介导的靶向人端粒酶反转录酶（htert）基因 rna 干扰（rnai）对白血病 Hl-6细胞 htert 基因、蛋白表达和细胞增殖的影响。方法采用靶向 htert 基因 rnai 的重组质粒 psilecer1.0-U6/htert 转染 Hl-60细胞,以空载体质粒 psilecer1.0-U6转染组、转染试剂及空白组作对照，将转染后24h、72h、120h 的细胞用 rt-Pcr 检测细胞 htert 基因 mrna 的表达，Western-blot 检测细胞 htert 蛋白的表达，Mtt 检测细胞增殖活性。结果转染psilecer1.0-U6/htert 质粒24h、72h 和120h 后，Hl-60细胞 htert 基因、蛋白表达水平均低于各对照组（p ＜0.05），细胞增殖抑制率高于各对照组（p ＜0.05）。htert 基因、蛋白和细胞增殖抑制率在转染 psilecer1.0-U6/htert 质粒24h、72h 和120h 组间无明显差异（p ＞0.05），在3个对照组间也无明显差异（p ＞0.05）。结论载体介导的靶向htert 基因的 rnai 技术能在体外抑制 Hl-60细胞 htert 基因和蛋白的表达，增加细胞凋亡，抑制增殖。
목적：탐토재체개도적파향인단립매반전록매（htert）기인 rna 간우（rnai）대백혈병 Hl-6세포 htert 기인、단백표체화세포증식적영향。방법채용파향 htert 기인 rnai 적중조질립 psilecer1.0-U6/htert 전염 Hl-60세포,이공재체질립 psilecer1.0-U6전염조、전염시제급공백조작대조，장전염후24h、72h、120h 적세포용 rt-Pcr 검측세포 htert 기인 mrna 적표체，Western-blot 검측세포 htert 단백적표체，Mtt 검측세포증식활성。결과전염psilecer1.0-U6/htert 질립24h、72h 화120h 후，Hl-60세포 htert 기인、단백표체수평균저우각대조조（p ＜0.05），세포증식억제솔고우각대조조（p ＜0.05）。htert 기인、단백화세포증식억제솔재전염 psilecer1.0-U6/htert 질립24h、72h 화120h 조간무명현차이（p ＞0.05），재3개대조조간야무명현차이（p ＞0.05）。결론재체개도적파향htert 기인적 rnai 기술능재체외억제 Hl-60세포 htert 기인화단백적표체，증가세포조망，억제증식。
Objective to explore the effect of vector medidated targeting human telomerase reverse transcriptase (htert) gene rna interfeerence (rnai) on the gene,protein expression and cell proliferetion of leukemia Hl-60 cells. Methods the targeting htert gene rnai recombination vector (psilecer1.0-U6/htert) was transfected into Hl-60 cells. the Hl-60 cells transfected with empty vector (psilecer1.0-U6), rnai-mate and Hl-60 cells were used as the controls. rt-Pcr was used to detect the expression of htert mrna on 24h, 72h, 120h after cell transfection, and Western-blot was used to detect the expression of htert protein. Mtt assay was used to observe the proliferative effects of cells. Results the expression levels of htert gene and protein in Hl-60 cells were lower and the proliferation inhibitory rate were higher than those of each control group after transfection with psilencer1.0-U6/ htert for 24 h, 72 h and 120h (P < 0.05).there were no differences among Hl-60 cells transfected with psilencer1.0-U6/ htert for 24 h, 72 h and 120h,and no differences were also observed among the three control groups(P > 0.05).Conclusion Vector-mediated targeting htert gene rnai can down-regulate the expression of htert gene and protein expression of htert of Hl-60, increase the cell apoptosis and inhibit cell proliferation.