MDM2反义寡核苷酸联合紫杉醇对乳腺癌MCF-7细胞株的作用

目的:探讨靶向MDM2反义寡核苷酸(ASON)联合紫杉醇对乳腺癌MCF-7细胞株的影响。方法:合成一段与MDM2 mRNA特异性结合的反义寡核苷酸和与反义寡核苷酸有4个碱基不同的的错义寡核苷酸(MON),脂质体2000介导不同浓度的MDM2ASON转染MCF-7乳腺癌细胞系,转染的乳腺癌细胞通过1μmol/L紫杉醇药物处理后,采用RT-PCR和Western Blot方法检测MDM2 ASON联合紫杉醇的协同作用及对乳腺癌MCF-7细胞株的抑制效率,MTT观察给药后MCF-7细胞的增殖能力和药物敏感性。结果:MDM2反义寡核苷酸联合紫杉醇明显下调MDM2 mRNA及MDM2蛋白表达水平,抑制MCF-7细胞的生长,随着MDM2 ASON浓度的增加,MDM2表达越来越低,协同作用越来越强,呈剂量依赖关系,A500联合紫杉醇的协同作用最明显,MTT显示紫杉醇处理的转染MCF-7细胞增殖抑制率明显增高,A500抑制增殖作用最明显,抑制率达(13.0±0.84)%。结论:不同浓度MDM2 ASON转染后的乳腺癌MCF-7细胞,等浓度紫杉醇处理后,乳腺癌MCF-7细胞MDM2表达明显降低,细胞凋亡增加,,MDM2 ASON联合紫杉醇对MCF-7细胞有协同作用,提高了乳腺癌MCF-7细胞对紫杉醇的药物敏感性。

Effects of MDM2 Antisense Oligonucleotide Combined with Paclitaxelon Human Breast Cancer Cells MCF-7

Objective: To investigate the effects of the MDM2 antisense oligonucleotide(ASON) combined with Paclitaxel on hu-man breast cancer cells MCF-7.Methods: The synthesis of antisense oligonucleotides specific binding of MDM2 mRNA and missense oligonucleotides(MON) different from four bases,different concentrations of MDM2 ASON mediated by Lipofectamine 2000 transfected MCF-7 breast cancer cell lines,breast cancer cells transfected by 1 μmol/L paclitaxel treatment.The expression of MDM2 mRNA and protein was determined by RT-PCR and Western blotting,To detect synergies of MDM2 ASON combined with paclitaxel and the inhibi-tion efficiency of breast cancer cells MCF-7,the proliferation of MCF-7 cell to paclitaxe and chemosensitivity were observed by MTT as-say.Results: The antisense oligonucleotide combined with Paclitaxel efficiently down-regulated MDM2 mRNA and protein expression,inhibit the growth of MCF-7 cells.MDM2 expression was getting lower and lower with the increase of the concentration of MDM2 ASON growing in a dose dependent relationship,the synergy of the A500 combined with paclitaxel was the most obvious.MTT showed that proliferation inhibition rate of MCF-7 cell transfected to paclitaxel increased significantly,A500 was the most significant effect,inhibi-tion rate was(13.0 ± 0.84)%.Conclusion: Human breast cancer cell MCF-7 transfected was treated by a concentration of paclitaxel,MDM2 expression was significantly decreased,increased apoptosis,MDM2 ASON combined with paclitaxel on MCF-7 cells had a syn-ergistic effect,improved the sensitivity of breast cancer MCF-7 cells to paclitaxel.