反义硫代磷酸寡核苷酸逆转肝癌细胞多药耐药的实验研究

REVERSAL OF MULTIDRUG RESISTANCE OF HUMAN HEPATOMA RESISTANT CELLS IN VITRO BY ANTISENSE PHOSPHOROTHIOATE OLIGONUCLEOTIDE

目的 观察反义硫代磷酸寡核苷酸 (ASOND)逆转耐药肝癌细胞的多药耐药作用。方法 用人工合成互补于mdrl基因 5′端AUG起始密码子的反义 2 0聚硫代磷酸寡核苷酸 ,以Lipofectamine为载体 ,转染人耐药肝癌细胞SMMC 772 1,MTT测定细胞对化疗药物的敏感性 ,通过RT PCR检测mRNA的表达 ,流式细胞仪分析P 170的表达。结果 ASOND可抑制SMMC 772 1细胞mRNA及P 170的表达 ,增加耐药细胞对化疗药物的敏感性 ,最佳ASOND作用浓度为 0 .5 μmol/L。 结论 ASOND可增加耐药细胞对化疗药物的敏感性 ,部分逆转耐药肝癌细胞耐药。

Objective To investigate the reversal effect of antisense phosphorothioate oligonucleotide (ASOND) on human hepatoma resistant cells. Methods Human hepatoma resistant cells SMMC-7721 was transfected with synthetic antisense phosphorothioate oligonucleotide complementary to the 5′ region flanking the AUG initiation codon mediated by lipofectamine. In vitro drug sensitivity was measured by MTT assay. The expression of P-170 was determined by flow cytometry and mRNA was assessed by RT-PCR. Results ASOND inhibited the expression of mRNA and p-170 in SMMC-7721, enhanced the sensitivity of SMMC-7721 to chemotherapeutic drug. The best inhibitory effect was achived by the dose of 0.5μmol/L. Conclusion ASOND enhanced the sensitivity of SMMC-7721 to chemotherapeutic drug and reversed the multidrug resistance of SMMC-7721 partially.