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目的：了解三螺旋形成寡核苷酸（ＴＦＯ）。反义寡核苷酸（ＯＤＮａｓ）抑制肝癌细胞生长的作用。方法：以Ｎ－ｒａｓ第二转录起始位点及翻译起始区１－６密码为靶点，合成１２聚硫代磷酸ＴＦＯ（ＴＦＯ１２）、１９聚硫代磷酸ＯＤＮａｓ（ＯＤＮａｓ１９），同时合成 １９聚无关序列硫代磷酸寡核苷酸对照（ＯＤＮｃｏｎ）。在人肝癌 Ｂｅｌ ７４０２细胞观察了硫代磷酸寡核苷酸对Ｎ－ｒａｓ表达ｐ２１蛋白的抑制及对细胞增殖的影响。结果： ＴＦＯ１２、 ＯＤＮａｓ１９处理的细胞ｐ２１合成降低了８０． ２％、６７．５％ ；细胞增殖降低了 ８８％、７７％；~３Ｈ－ＴｄＲ掺入降低了 ７８．２％、６９．５％。 ＴＦＯ１２、ＯＤＮａｓ１９的生长抑制作用呈剂量依赖性，该作用在处理后６小时即可检出，１２小时达高峰，以后逐渐降低，但７２小时后仍有一定作用。两者联合应用抑制作用增强。ＯＤＮｃｏｎ 对Ｎ－ｒａｓ表达ｐ２１及细胞增殖无影响。结论：针对Ｎ－ｒａｓ不同基因区的ＴＦＯ、ＯＤＮａｓ能有效抑制ｒａｓ表达及ｒａｓ相关性肝癌细胞的增殖，具有潜在应用价值。
Objective: To understand triple helix forming oligonucleotide (TFO). Antisense oligonucleotides (ODNas) inhibit the growth of hepatoma cells. METHODS: The 12-polythiophosphorothioate TFO (TFO12) and 19-polythiophosphoric acid ODNas (ODNas19) were synthesized using the N-ras second transcription initiation site and the translation initiation region 1-6 code as targets. Poly-independent sequence phosphorothioate oligonucleotide control (ODNcon). In human liver cancer Bel 7402 cells, the inhibition of N-ras expression of p21 protein by phosphothioate oligonucleotides and the effect on cell proliferation were observed. RESULTS: The p21 synthesis of cells treated with TFO12 and ODNas19 was reduced by 80. 2%, 67.5%; cell proliferation decreased by 88% and 77%; ~3H-TdR incorporation decreased by 78.2% and 69.5%. The growth inhibitory effects of TFO12 and ODNas19 were dose-dependent. The effect was detected within 6 hours after treatment, peaked at 12 hours, and then gradually decreased, but still had a certain effect after 72 hours. The combined use of the two has enhanced inhibition. ODNcon had no effect on N-ras expression p21 and cell proliferation. Conclusion: TFO and ODNas targeting different N-ras gene regions can effectively inhibit the expression of ras and the proliferation of ras-associated hepatocellular carcinoma cells, which has potential application value.