CAJ | 학술논문

目的构建人端粒酶反转录酶(HTERT)启动子调控的NK4基因重组腺病毒表达系统,探讨NK4基因对结肠癌生长、侵袭转移的抑制作用.方法以Ad HTERTp-NK4重组腺病毒载体感染结肠癌细胞HTC116,分别以RT-PCR方法和Western blot检测细胞内NK4基因mRNA及蛋白表达量:MTT和体外侵袭实验观察NK4对结肠癌细胞增殖及侵袭转移能力的抑制作用.建立结肠癌裸鼠皮下移植瘤模型,观察重组腺病毒Ad HTERTp-NK4对裸鼠结肠癌移植瘤生长的影响.结果 Ad HTERTp-NK4重组腺病毒载体感染HCT116细胞后,HCT116细胞内NK4 mRNA及蛋白表达水平明显增高:Ad HTERTp-NK4重组腺病毒感染后,HCT116细胞生长抑制率和侵袭抑制率分别为47.14%和40.63%,与Ad HTERTp-GFP(空载体)组(2.75%和2.31%)比较,差异均有统计学意义(均P＜0.05).局部注射Ad HTERTp-NK4重组腺病毒载体后,结肠癌裸鼠皮下移植瘤模型肿瘤生长受抑,抑瘤率为47.3%,与空载体组(4.6%)比较,差异有统计学意义(P＜0.05).结论 HTERT启动子调控NK4基因靶向治疗系统可有效抑制结肠癌的增殖及侵袭转移,在结肠癌的基因治疗具有较好的应用前景.
목적 구건인단립매반전록매(HTERT)계동자조공적NK4기인중조선병독표체계통,탐토NK4기인대결장암생장、침습전이적억제작용.방법 이Ad HTERTp-NK4중조선병독재체감염결장암세포HTC116,분별이RT-PCR방법화Western blot검측세포내NK4기인mRNA급단백표체량:MTT화체외침습실험관찰NK4대결장암세포증식급침습전이능력적억제작용.건립결장암라서피하이식류모형,관찰중조선병독Ad HTERTp-NK4대라서결장암이식류생장적영향.결과 Ad HTERTp-NK4중조선병독재체감염HCT116세포후,HCT116세포내NK4 mRNA급단백표체수평명현증고:Ad HTERTp-NK4중조선병독감염후,HCT116세포생장억제솔화침습억제솔분별위47.14%화40.63%,여Ad HTERTp-GFP(공재체)조(2.75%화2.31%)비교,차이균유통계학의의(균P＜0.05).국부주사Ad HTERTp-NK4중조선병독재체후,결장암라서피하이식류모형종류생장수억,억류솔위47.3%,여공재체조(4.6%)비교,차이유통계학의의(P＜0.05).결론 HTERT계동자조공NK4기인파향치료계통가유효억제결장암적증식급침습전이,재결장암적기인치료구유교호적응용전경.
Objective To investigate anti-tumor effect of the recombined adenovirus encoding NK4 gene regulated by human telomerase reverse transcriptase (HTERT) promoter (Ad HTERTp-NK4) on human colon cancer.Methods Colon cell line HCT116 was infected with Ad HTERTp-NK4.NK4 expression was determined by RT-PCR and Western blot.The cell-growth inhibition rate and the invasive capacity of cells were evaluated by MTT method and reconstituted basement membrane invasion assay.The model of subcutaneous tumor was generated by injection of HCT116 cells into the dorsum of nude mice.Ad HTERTp-NK4 was injected around the tumor tissues,and tumor growth was observed.Results NK4 gene was highly expressed in infected HCT116 cells.The cell growth inhibition rate and the invasive inhibition rate in Ad HTERTp-NK4 cells were 47.14% and 40.63% respectively,which were significantly higher than those in the control cells (2.75% and 2.31%,P＜0.05).Tumor growth was significantly inhibited in mice injected with Ad HTERTp-NK4,and the tumor growth inhibition rate was 47.3%,which was significantly higher than that in the controls (4.6% ,P＜0.05).Conclusion Ad HTERTp-NK4 can inhibit tumor growth and decrease the invasive capacity of tumor cells,which makes it an ideal candidate for new gene therapy for colon carcinoma.