Tankyrase1 反义核酸干预肺癌移植瘤生长的实验研究*

目的:观察Tankyrase1（端锚聚合酶1，TANK 1）的正义（TANK 1-SODN）及反义寡核苷酸（TANK 1-ASODN ）作用下人肺癌细胞移植瘤的生长情况，探讨TANK 1-ASODN 抑制癌细胞增殖的实际效果及作用机制，为肺癌基因治疗提供新线索。方法:将14只荷瘤裸鼠随机分成3 组，每日一次瘤体多位点分别注射相应剂量TANK 1-SODN、TANK 1-ASODN 及生理盐水。15天后，观察各组移植瘤体积、组织学及电镜下肺癌细胞超微结构的改变，使用链酶亲和素- 酶复合物（SABC）免疫组化法及原位杂交法检测3组移植瘤中端粒酶反转录酶（hTERT ）蛋白及其mRNA 表达水平。结果:TANK 1-ASODN 组移植瘤体积明显小于TANK 1-SODN及对照组，差异有显著性统计学意义（P<0.01），而TANK 1-SODN与对照组肿瘤体积差异无显著性统计学意义（P>0.01）。 电镜证实TANK 1-ASODN 通过多种途径，有明显的癌细胞杀灭效果。此外，TANK 1-ASODN 组癌细胞hTERT 蛋白及其mRNA 表达水平明显低于其他两组，差异有显著性统计学意义（P 均<0.01）。 结论:TANK 1-ASODN 明显降低了肺癌细胞中hTERT 蛋白及其mRNA的高水平表达，促进癌细胞坏死及凋亡，显著抑制了肺癌细胞增殖。 

The Inhibitory Effect of Antisense Oligodeoxynucleotides for Tankyrase 1 on Human Lung Cancer Cell Nodules

Objective: To observe the effect of sense and antisense oligodeoxynucleotides for tankyrase 1 (TANK 1-SODN and TANK 1-ASODN) on murine tumor growth and then to investigate the suppression mechanism of TANK 1-ASODN on cancer cell proliferation. Methods:Fourteen nude mice with lung cancer nodules were randomly divided into 3 groups. Multiple direct intratumoral injections of synthesized TANK1-ASODN were given for 15days. Mice treated with TANK 1-SODN and saline were used as the control groups. Tumor volume was measured and histopathological and ultrastructural characteristics of the tumor tissue were observed through microscopy and electron microscopy. SABC immunohistochemistry and in situ hybridization were used to detect the expression of hTERT telomerase protein and mRNA.Results: After15days of treatment, tumor size in the TANK1-ASODN treated group was much smaller than in the other 2 groups ( P<0.01). Electron microscopy images showed that TANK 1-ASODN had an apparent killing effect on lung cancer cells. Moreover, a statistically significant decrease in the positive expression ratio of telomerase hTERT protein and Mrna (P<0.01) was observed in the TANK1-ASODN group. Conclusion:TANK 1-ASODN can decrease the expression of hTERT, promote tumor cell degeneration and necrosis, and suppress proliferation of human lung cancer cells.