不同修饰型反义寡核苷酸抑制癌基因表达生物学效应的初步观察

Observation of Inhibitory Effect of Different Modified Type Antisense Oligonucleotides on Gene Expression

比较了两种不同的反义寡核苷酸抑制ＨＬ－６０细胞ｃ－ｍｙｃ癌基因表达的效应。二者有相同的碱基序列，但有不同的修饰方式，即全硫代修饰及卟啉修饰；另外还合成了两种非修饰型的反义和错义寡核苷酸作为对照。ＭＴＴ细胞增殖实验、ｃ－ｍｙｃｍＲＮＡ的Ｄｏｔｂｌｏｔ检测、ｃ－ｍｙｃ蛋白的免疫组化检测等结果显示卟啉修饰的寡核苷酸除了具有明显的的特异性抑制基因表达的效果外。

Two different modified type antisense oligonucleotides complemetary to c myc mRNA were compared in inhibitory effect on gene targeted expression and cell proliferation One was the phosphorothioate oligonucleotide(S O) and the other contained a porphyrin group(P O) At the same time,a mis sense oligomer ( M O ) and an anti sense oligomer(A O) as the controls were also synthesized Human acute promyelocyte leukemia cell line (HL 60), which can express high level of c myc oncoprotein, was chosen as the target cell The above four oligomers were incubated with HL 60 cells for 4, 24, 48,72 h respectively The results from dot blot hybridizaton for amount of c myc mNA level, MTT spectrophotometry for determination of HL 60 cell proliferation and immuno histochemistry reaction for amount of c myc protein showed that P O modified oligonucleotides indeed inhibited the expression of c myc oncogene in HL 60 cells and increased the additional effect of cell death