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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。
Objective: To investigate the effect of targeting MDM2 antisense oligonucleotide (ASON) and paclitaxel on breast cancer MCF-7 cell line. METHODS: An antisense oligonucleotide specifically binding to MDM2 mRNA and a missense oligonucleotide (MON) different from antisense oligonucleotide by 4 bases were synthesized. Lipofectamine 2000 mediated different MDM2ASON was transfected into MCF-7 breast cancer cell line. The transfected breast cancer cells were treated with 1μmol / L paclitaxel, and the synergistic effect of MDM2 ASON and paclitaxel was detected by RT-PCR and Western Blot. 7 cell line, MTT was used to observe the proliferation and drug sensitivity of MCF-7 cells after administration. Results: MDM2 antisense oligodeoxynucleotides combined with paclitaxel significantly down-regulated the expression of MDM2 mRNA and MDM2 protein and inhibited the growth of MCF-7 cells. With the increase of MDM2 ASON concentration, the expression of MDM2 was lower and lower and the synergistic effect was stronger and stronger , The synergistic effect of A500 and paclitaxel was the most obvious. MTT showed that paclitaxel-treated MCF-7 transfected MCF-7 cells were significantly increased, and A500 inhibited the proliferation significantly (13.0 ± 0.84)%. CONCLUSION: The MDM2 expression in breast cancer MCF-7 cells is significantly decreased and the apoptosis is increased after treated with different concentration of MDM2 ASON in breast cancer MCF-7 cells treated with paclitaxel of equal concentration. MDM2 ASON combined with paclitaxel has a significant effect on MCF-7 cells Synergy, increased breast cancer MCF-7 cells to paclitaxel drug sensitivity.