| 韩 雪,方 红,聂 琛.干扰ANXA3基因表达对乳腺癌细胞增殖和凋亡的影响[J].肿瘤学杂志,2019,25(7):622-626. |
| 干扰ANXA3基因表达对乳腺癌细胞增殖和凋亡的影响 |
| Effect of Interfering ANXA3 Gene Expression on Proliferation and Apoptosis of Breast Cancer Cells |
| 投稿时间：2018-01-02 |
| DOI：10.11735/j.issn.1671-170X.2019.07.B007 |
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| 中文关键词: 乳腺癌 增殖 凋亡 ANXA3 |
| 英文关键词:breast cancer proliferation apoptosis ANXA3 |
| 基金项目: |
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| 摘要点击次数: 1874 |
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| 中文摘要: |
| 摘 要：［目的］ 探究干扰膜联蛋白A3（ANXA3）基因表达对乳腺癌细胞增殖、凋亡的影响以及可能作用机制。［方法］ 以乳腺癌MCF-7细胞为研究对象，RNA干扰技术沉默ANXA3基因表达，荧光定量RT-PCR和Western Blot检测转染后细胞中ANXA3 mRNA及蛋白表达；四甲基偶氮唑（MTT）法检测细胞的增殖能力变化，膜联蛋白 V-FITC（Annexin V-FITC）/碘化丙啶(PI)双染法检测细胞的凋亡率，蛋白质印迹法（Western Blot）检测细胞中B细胞淋巴瘤/白血病-2（Bcl-2）、Bcl-2相关X蛋白(Bax)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3（Cleaved Caspase-3）蛋白的水平。［结果］ 空白对照组、阴性对照组和ANXA3干扰组细胞中ANXA3 mRNA的表达量分别为0.078±0.006、0.072±0.007和0.008±0.001；ANXA3蛋白表达量分别为0.792±0.085、0.764±0.078和0.353±0.039；细胞的凋亡率分别为（3.425±0.643）%、（3.537±0.740）%、（23.455±3.686）%；Bcl-2蛋白水平分别为1.256±0.247、1.239±0.265、0.624±0.065；Bax蛋白水平分别为0.856±0.086、0.842±0.079、1.526±0.344；Cleaved Caspase-3蛋白水平分别为0.522±0.057、0.535±0.062、1.230±0.367。与空白对照组相比，ANXA3干扰组细胞中ANXA3 mRNA和蛋白水平显著性降低（P<0.05），细胞的增殖能力显著性降低（P<0.05），其凋亡率显著性增加（P<0.05）；细胞中Bcl-2蛋白水平显著性下调（P<0.05），Bax、Cleaved Caspase-3蛋白表达量显著性上调（P<0.05）。［结论］ 干扰ANXA3表达抑制乳腺癌细胞增殖，诱导其凋亡，其作用与Bcl-2、Bax、Cleaved Caspase-3蛋白水平变化引起的细胞凋亡通路的变化有关。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the effect of interfering Annexin A3（ANXA3） gene expression on proliferation and apoptosis of breast cancer cells and its mechanism. ［Methods］ The ANXA3 gene expression in breast cancer MCF-7 cells was silenced by RNA interference technique. The expressions of ANXA3 mRNA and protein were detected by RT-PCR and Western Blot，respectively. Tetramethylzole(MTT) method was used to examine the cell proliferation and the apoptotic rate was determined by Annexin V-FITC/PI staining method. The expression of B cell lymphoma/lewkmia-2（Bcl-2），Bcl-2-related protein X(Bax)，cleaved cysteinyl aspartate specific proteinase 3（cleaved Caspase-3）in MCF-7 cells were detected by Western blot. ［Results］ The expression levels of ANXA3 mRNA were 0.078±0.006，0.072±0.007，0.008±0.001 in blank control group，negative control group and ANXA3 interference group，respectively；and the protein expression levels were 0.792±0.085，0.764±0.078 and 0.353±0.039. The apoptosis rates were（3.425±0.643）%，（3.537±0.740）%，（23.455±3.686）%，respectively，the protein expression levels of Bcl-2 were 1.256±0.247，1.239±0.265，0.624±0.065，respectively；those of Bax were 0.856±0.086，0.842±0.079，1.526±0.344，respectively；and those of cleaved Caspase-3 were 0.522±0.057，0.535±0.062，1.230±0.367 in three groups，respectively. Compared with the blank control group，the expression levels of ANXA3 mRNA and protein were significantly decreased(P<0.05)，cell proliferation was significantly decreased(P<0.05)，and the apoptosis rate was significantly increased(P<0.05). The protein expression levels of Bcl-2 were significantly down-regulated(P<0.05)，and the protein expression levels of Bax and cleaved Caspase-3 were significantly up-regulated(P<0.05). ［Conclusion］ Interference of ANXA3 expression inhibits the proliferation and induces apoptosis of breast cancer MCF-7 cells，which is related to the change in the expression levels of Bcl-2，Bax and cleaved Caspase-3. |
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