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为了探讨HSP70对癌细胞的增生及生存的作用,我们使用自己合成的S(硫)修饰的HSP70反义寡脱氧核苷酸(an-tisense oligomer)阻断癌细胞的 HSP70基因的表达,分析了HSP70反义寡脱氧核苷酸的抗肿瘤作用中细胞凋亡与HSP70的关系.我们合成的抗核酸酶的S(硫)修饰的反义脱氧寡核苷酸(碱基序列为5’-CGCGGCTTTGGCCAT-3’)与人HSP7OmRNA的起始密码子以及其下游的四个密码子互补.此外,合成了相应的正义脱氧寡核苷酸(sense oligomer,5’-ATGGCCAAAGCC GCG-3’)以及无义脱氧寡核苷酸(nonsenseoligomer,5’-CGGGTATGCTTC GCC-3’)作为对照.这些寡脱氧核苷酸经高压液相色谱及反相色谱纯化,分析其细胞内代谢动力学,发现90～180分钟之间可进入细胞内,并且在其细胞内可稳定存在24/小时以上.经 Westem blor、定量免疫荧光以及Northem blot分析发现HSP70反义脱氧寡核苷酸
To investigate the effect of HSP70 on the proliferation and survival of cancer cells, we used an S-(thio)-modified HSP70 antisense oligodeoxynucleotide (an-tisense oligomer) to block the expression of HSP70 gene in cancer cells. Relationship between apoptosis and HSP70 in HSP70 antisense oligodeoxynucleotide anti-tumor effect. We synthesize nuclease-resistant S(thio)-modified antisense deoxyoligonucleotides (base sequence 5’-CGCGGCTTTGGCCAT -3’) is complementary to the start codon of human HSP7O mRNA and its four downstream codons. In addition, the corresponding sense oligomer (5’-ATGGCCAAAGCC GCG-3’) and nonsense are synthesized. The nonsenseoligomer (5’-CGGGTATGCTTC GCC-3’) was used as a control. These oligodeoxynucleotides were purified by high pressure liquid chromatography and reversed phase chromatography to analyze the intracellular metabolic dynamics and found to be 90-180 minutes. Into the cell between, and in its cells can be stable for more than 24/hour. HSP70 antisense deoxyribonucleotide was found by Westem blor, quantitative immunofluorescence and Northern blot analysis