反义hTERT对人舌鳞癌细胞裸鼠体内成瘤的抑制作用的研究

Inhibitory effect of antisense oligodeoxynucleotides against hTERT on nude mice transplantation tumor of human tongue Tca8113 cancer cell

目的观察人端粒酶逆转录酶(human telomerase reverse transcriptase,hTERT)基因反义寡核苷酸对人舌鳞癌Tca8113细胞株裸鼠体内成瘤的影响及其作用机制。方法实验设反义寡核苷酸组、正义寡核苷酸组、空载体组和空白对照组,后3个组为对照组。采用RT-PCR法检测hTERT基因表达变化,TRAP法测定端粒酶活性,免疫组化法检测细胞增殖核抗原(PCNA)表达量。所有数据用单因素ANOVA方差分析进行统计学检验。结果hTERT基因反义寡核苷酸组人舌鳞癌Tca8113细胞体内成瘤能力、瘤体体积明显小于对照组,hTERTmRNA和端粒酶活性表达低于对照组(P<0.05)。结论hTERT基因反义寡核苷酸能够抑制人舌鳞癌Tca8113体内成瘤,抑制细胞hTERT基因的转录和端粒酶活化可能是其作用机制之一。

Objective To study the effect on nude mice transplantation tumors of human tongue Tca8113 cancer cell and its mechanisms, affected by antisense oligodeoxynucleotides （ASODN） against human telomerase reverse transcriptase （hTERT）, and thus might be used to illustrating the significance of antisense hTERT in treating the squamous cell of human tongue. Methods ASODN was designed complementary to target sites of hTERT mRNA. Sense oligodeoxynucleotides（SODN）, blank carrier,blank culture medium were control groups. The Tca8113 cells were transfected once time with ASODNs, and the influences on transplantation tumor growth, hTERT mRNA, and telomerase activity were examined. The data were analyzed by one-way ANOVA of SPSS 11. 0 for windows. Results An distinctive reduction of Tca8113 cell viability in nude mice was achieved by treatment with ASODN. Additionally ,significant inhibition of proliferation （tumor foundation time ,tumor size,PCNA）, as well as an induction of hTERT mRNA were observed. As a consequence, the telomerase activity was inhibited by anti hTERT treatment. Conclusion hTERT ASODN causes remarkable inhibitory effects on the growth of human tongue Tca8113 cancer cells in nude mice and can differentially inhibit hTERT mRNA, consequently disables telomerase activity and thereby terminates the unlimited ability of cancer cell proliferation in vivo. hTERT ASODN represents a promising new treatment option of tongue tumors.