紫杉醇诱导三阴性乳腺癌细胞自噬相关蛋白LC3的表达及其作用
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黄俊辉, Email: 808003@csu.edu.com

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湖南省自然科学基金资助项目(11jj3122);中南大学研究生自主探索创新基金资助项目(2016zzts531)。


Paclitaxel induced expression of autophagy-associated protein LC3 in triple negative breast cancer cells and its action
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    摘要:

    目的:探讨紫杉醇对三阴性乳腺癌(TNBC)细胞自噬相关蛋白LC3表达的影响及意义。 方法:用CCK-8法测定紫杉醇对TNBC细胞MDA-MB-231增殖的抑制作用及25%抑制浓度(IC25);用IC25浓度的紫杉醇作用MDA-MB-231细胞后,分别用免疫荧光化学法、Western blot、流式细胞术检测细胞LC3与凋亡相关蛋白的表达及细胞的凋亡率。 结果:紫杉醇呈浓度依懒性抑制MDA-MB-231细胞的增殖(P<0.05),其IC25为3.11 μg/mL。IC25浓度的紫杉醇处理后,MDA-MB-231细胞后LC3的表达量以及LC3B/LC3A比例明显升高、凋亡蛋白Bax与caspase-3蛋白表达量明显降低、总凋亡率与早期调亡率均明降低(均P<0.05)。 结论:紫杉醇可诱导TNBC细胞自噬相关蛋白LC3表达升高,该作用可能降低细胞凋亡,从而导致TNBC细胞产生紫杉醇耐药。

    Abstract:

    Objective: To investigate the influence of paclitaxel on expression of autophagy-associated protein LC3 in triple negative breast cancer (TNBC) cells and the significance. Methods: The inhibitory effect of paclitaxel on proliferation of TNBC MDA-MB-231 cells was determined by CCK-8 assay and then the 25% inhibition concentration (IC25) value was calculated. In MDA-MB-231 cells after treatment with IC25 concentration of paclitaxel, the expressions of LC3 protein and apoptosis-associated proteins as well as the cell apoptosis rates were determined by immunofluorescence histochemistry, Western blot analysis and flow cytometry, respectively. Results: The proliferation effect of TNBC-MDA-MB-231 cells was significantly inhibited by paclitaxel in a concentration-dependent manner (P<0.05), with an IC25 value of 3.11 μg/mL. In MDA-MB-231 cells after treatment with 3.11 μg/mL paclitaxel, the expression level of LC3 protein as well as LC3B/LC3A ratio were significantly increased, the expression levels of the apoptosis-related protein Bax and caspase-3 were significantly decreased, and the total and early apoptosis rates were significantly decreased (all P<0.05). Conclusion: Paclitaxel can induce the increased expression of the autophagy-related protein LC3, and this action may probably reduce the cell apoptosis and thereby cause paclitaxel resistance in TNBC cells.

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吴至佛|姚晓艺|汪灵|邓增艳|石静|黄俊辉.紫杉醇诱导三阴性乳腺癌细胞自噬相关蛋白LC3的表达及其作用[J].中国普通外科杂志,2018,27(6):732-739.
DOI:10.3978/j. issn.1005-6947.2018.06.012

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  • 收稿日期:2018-03-13
  • 最后修改日期:2018-05-19
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  • 在线发布日期: 2018-06-15