CAJ | 학술논문

【目的】探讨人端粒酶启动子（hT ERT ）联合正常组织特异性miR‐34a （M IR）的整合靶向系统（hT ERT‐M IR）对胶质瘤细胞靶向性的影响。【方法】首先检测hT ERT、miR‐34a在人胶质瘤细胞株 U87、H4和人胶质细胞株HEB（正常细胞株）中的表达；通过质粒构建分别获得 hTERT‐Luc、hTERT‐MIR‐Luc、hTERT‐Bax 以及hTERT‐MIR‐Bax表达质粒，将hTERT‐Luc、hTERT‐MIR‐Luc质粒分别转染U87、H4和 HEB细胞中，检测三组细胞中hT ERT 与hT ERT‐M IR载体的表达活性；将hT ERT‐Bax以及hT ERT‐M IR‐Bax质粒分别转染U87、H4和HEB细胞中，比较三组细胞中hTERT‐Bax和hTERT‐MIR‐Bax对胶质瘤细胞增殖抑制能力的影响。【结果】hTERT在人胶质瘤细胞株U87和H4中特异性高表达，miR‐34a在人胶质细胞株HEB中特异性高表达；在正常胶质细胞株 HEB中，hTERT‐MIR的活性较 hTERT 明显减低（ P ＜0．01）；在胶质瘤细胞株 U87和 H4h中TERT‐Bax和hTERT‐MIR‐Bax 对细胞增殖抑制比较差异无显著性，但在正常胶质细胞株 HEB中，hTERT‐M IR‐Bax对细胞的增殖抑制明显小于hT ERT‐Bax。【结论】hT ERT‐M IR是胶质瘤基因靶向治疗的有效载体系统。
【목적】탐토인단립매계동자（hT ERT ）연합정상조직특이성miR‐34a （M IR）적정합파향계통（hT ERT‐M IR）대효질류세포파향성적영향。【방법】수선검측hT ERT、miR‐34a재인효질류세포주 U87、H4화인효질세포주HEB（정상세포주）중적표체；통과질립구건분별획득 hTERT‐Luc、hTERT‐MIR‐Luc、hTERT‐Bax 이급hTERT‐MIR‐Bax표체질립，장hTERT‐Luc、hTERT‐MIR‐Luc질립분별전염U87、H4화 HEB세포중，검측삼조세포중hT ERT 여hT ERT‐M IR재체적표체활성；장hT ERT‐Bax이급hT ERT‐M IR‐Bax질립분별전염U87、H4화HEB세포중，비교삼조세포중hTERT‐Bax화hTERT‐MIR‐Bax대효질류세포증식억제능력적영향。【결과】hTERT재인효질류세포주U87화H4중특이성고표체，miR‐34a재인효질세포주HEB중특이성고표체；재정상효질세포주 HEB중，hTERT‐MIR적활성교 hTERT 명현감저（ P ＜0．01）；재효질류세포주 U87화 H4h중TERT‐Bax화hTERT‐MIR‐Bax 대세포증식억제비교차이무현저성，단재정상효질세포주 HEB중，hTERT‐M IR‐Bax대세포적증식억제명현소우hT ERT‐Bax。【결론】hT ERT‐M IR시효질류기인파향치료적유효재체계통。
[Objective]To explore a novel vector system based on hTERT promoter and cell‐specific miR‐34a for targeted therapy of glioma .[Methods] The expression levels of hTERT and miR‐34a were detected by quanti‐tative real‐time polymerase chain reaction (qRT‐PCR) in U87 and H4 glioma and normal colloid cells .The vectors of hTERT‐Luc ,hTERT‐MIR‐Luc ,hTERT‐Bax and hTERT‐MIR‐Bax were constructed and transfected into U 87 and H4 glioma and normal colloid cells respectively .The expression activities of hTERT and hTERT‐MIR vectors were detected by luciferase assay in U 87 and H4 glioma and normal colloid cells .And the proliferative capacities of U87 and H4 glioma and normal colloid cells regulated by hTERT‐Bax and hTERT‐MIR‐Bax were evaluated by thi‐azolyl blue tetrazolium blue (MTT) .[Results]The expression of hTERT promoter was stronger in U87 and H4 cells and miR‐34a was highly expressed in normal colloid cells .After incorporating normal cell‐specific miRNA‐targeted sequences ,the activity of hTERT‐MIR system decreased in normal cells ( P <0 .01) .Furthermore ,the proliferative capacity of glioma cells after transfection showed no significant differences .However ,significant differences existed in normal colloid cells .[Conclusion]hTERT‐MIR is an effective vector system of targeted gene therapy for glioma .