| 陈园园,叶园园,李志臻.SALL4基因沉默对MCF-7/A细胞增殖和化疗敏感性的影响[J].中国肿瘤,2016,25(2):132-137. |
| SALL4基因沉默对MCF-7/A细胞增殖和化疗敏感性的影响 |
| Effect of SALL4 Gene Silencing on the Proliferation and Drug Sensitivity of Multidrug Resistant Breast Cancer Cell MCF-7/A |
| 投稿时间：2015-10-10 |
| DOI：10.11735/j.issn.1004-0242.2016.02.A011 |
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| 中文关键词: SALL4基因 乳腺肿瘤 表柔比星 |
| 英文关键词:SALL4 breast neoplasms epirubicin |
| 基金项目:国家自然科学基金 (81172078)；上海交通大学医工交叉基金项目(YG2013MS16) |
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| 中文摘要: |
| 摘 要：［目的］ 探讨沉默SALL4基因对乳腺癌耐药细胞MCF-7/A的增殖及化疗敏感性的影响。［方法］ qRT-PCR和Western Blot实验检测MCF-7、MCF-7/A细胞内SALL4表达水平，运用慢病毒包装shRNA方法沉默MCF-7/A细胞的SALL4基因；实验组（Lv-shSALL4）、阴性对照组（Lv-shNC）和空白对照组(CON)。CCK8法、克隆形成实验检测SALL4沉默对细胞增殖能力的影响；CCK8法检测表柔比星处理细胞24h和48h的半数抑制浓度（IC50）；流式细胞仪检测细胞凋亡。［结果］ SALL4在MCF-7/A细胞中的表达水平显著高于MCF-7（P<0.05）。MCF-7/A细胞中SALL4基因被成功沉默，病毒感染MCF-7/A细胞72h后，细胞感染效率约95%，沉默SALL4基因后MCF-7/A细胞内SALL4 mRNA及蛋白表达均明显降低（P<0.05）。沉默MCF-7/A细胞的SALL4基因，显著抑制了细胞的增殖能力（P<0.05），增强了细胞对表柔比星的敏感性；表柔比星24h和48h的IC50均明显降低（P<0.05），并且表柔比星诱导的细胞凋亡增加（P<0.05）。［结论］ 沉默SALL4基因能够抑制乳腺癌耐药细胞MCF-7/A的增殖，增强耐药细胞对表柔比星的敏感性。SALL4基因沉默可能是乳腺癌治疗的新靶点。 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the effect of SALL4 gene silencing on the cellular proliferation and sensitivity of human breast cancer cell line MCF-7/A to chemotherapy. ［Methods］ The mRNA and protein levels of SALL4 in MCF-7 and MCF-7/A cells were examined by qRT-PCR and Western Blot，respectively. The lentivirus-mediated shRNA approach was applied to silence the SALL4 gene of MCF-7/A cells. The experiment set up an experimental group (Lv-shSALL4)，the negative control group (Lv-shNC) and blank control group(CON).The inhibitory effect on proliferation of MCF-7/A cells were detected by CCK8 assay and colony formation. Groups of CON，Lv-shNC and Lv-shSALL4 were cultivated with various concentrations of Epirubicin for 24h and 48h. Then，the IC50 values(the concentration of drug inhibiting 50% of the cells) of MCF-7/A at 24h and 48h were measured by CCK8 regent kit and the cell apoptosis rate was detected by flow cytometry. ［Results］ SALL4 obviously up-regulated in MCF-7/A cells，compared with to MCF-7 cells(P<0.05). SALL4 gene in MCF-7/A cells was successfully silenced by the lentivirus-mediated shRNA interference approach. After being transfected for 72h，the infection efficiency was about 95%.And both mRNA and protein levels of SALL4 in Lv-shSALL4 group were significantly reduced (P<0.05).CCK8 assay based growth curve and colony formation assay indicated that silencing SALL4 significantly inhibited the proliferation of MCF-7/A cells(P<0.05).Besides，down-regulation SALL4 markedly increased the sensitivity of MCF-7/A cells to Epirubicin.IC50 value of Epirubicinin Lv-shSALL4 group was significantly lower at 24h and 48h based on CCK8 assay(P<0.05).The cell apoptosis rate was higher in Lv-shSALL4 group than that in CON group after cultivating with Epirubicin for 48h(P<0.05).［Conclusion］ Silencing the SALL4 gene in MCF-7/A can inhibit the proliferation and enhance the chemotherapy sensitivity of drug resistant breast cancer cells MCF-7/A，towards Epirubicin.It suggests that SALL4 may be a promising target for the therapy of breast cancer. |
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