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目的研究联合应用靶向survivin的反义寡核苷酸(ASODN)和三苯氧胺(TAM)对MCF-7乳腺癌细胞的作用。方法将一条20mer的ASODN序列与TAM作用于MCF-7乳腺癌细胞,分别为TAM组(单独应用TAM)、ASODN组(单独应用ASODN)及TAM+ASOND联合组(联合应用TAM+ASODN).运用四甲基偶氮唑盐比色试验(MTT)、流式细胞仪(FCM)、原位杂交以及分光光度法分别检测各组MCF-7细胞的抑制率、细胞周期和凋亡率、survivin mRNA表达和caspase-3活性。结果 TAM+ASOND联合组对MCF-7细胞的抑制率〔(62.26±3.92)%〕明显高于TAM组〔(42.30±6.63)%〕及ASODN组〔(54.77±9.99)%〕,P<0.05.TAM+ASOND联合组的细胞凋亡率为(28.08±4.32)%,明显高于TAM组〔(18.94±4.01)%〕及ASODN组〔(21.12±3.95)%〕,P<0.01.TAM+ASODN联合组对MCF-7细胞的G0/G1期阻滞作用明显强于TAM组和ASODN组(P<0.05,P<0.01);TAM+ASODN联合组survivin mRNA表达阳性率为(13.38±3.45)%,明显低于TAM组〔(39.67±7.42)%〕或ASODN组〔(27.50±5.80)%〕,P<0.01.TAM+ASOND联合组MCF-7细胞的caspase-3活性(0.93±0.13)高于TAM组(0.50±0.09)或ASODN组(0.64±0.08),P<0.01.结论靶向survivin的反义寡核苷酸能够促进MCF-7乳腺癌细胞凋亡,增加其对TAM治疗的敏感性。
Objective To investigate the effect of combined use of antisense oligonucleotides targeting survivin (ASODN) and tamoxifen (TAM) on MCF-7 breast cancer cells. Methods A 20-mer ASODN sequence and TAM were administered to MCF-7 breast cancer cells, which were TAM group (TAM alone), ASODN group (ASODN alone) and TAM + ASOND combined group (TAM + ASODN group) MTT assay, flow cytometry (FCM), in situ hybridization and spectrophotometry were used to detect the inhibition rate, cell cycle and apoptosis rate, survivin mRNA Expression and caspase-3 activity. Results The inhibition rate of MCF-7 cells in TAM plus ASOND group was significantly higher than that in TAM group (62.26 ± 3.92% vs 42.30 ± 6.63%, P <0.05), and (54.77 ± 9.99)% in ASODN group (28.08 ± 4.32)% in TAM plus ASOND group was significantly higher than that in TAM group (18.94 ± 4.01)% and (21.12 ± 3.95)% in ASODN group, P <0.01.TAM + ASODN combined group had significantly higher G0 / G1 phase arrest than that of TAM group and ASODN group (P <0.05, P <0.01); The positive rate of survivin mRNA in TAM + ASODN combined group was (13.38 ± 3.45) %, Which was significantly lower than that of TAM group (39.67 ± 7.42%) or ASODN group (27.50 ± 5.80%), P <0.01. The caspase-3 activity of MCF- (0.50 ± 0.09) or ASODN group (0.64 ± 0.08), P <0.01.Conclusion Antisense oligonucleotides targeting survivin can promote the apoptosis of MCF-7 breast cancer cells and increase their effect on TAM-treated Sensitivity.