丝胶靶向Akt调控PI3K/Akt/mTOR信号通路诱导三阴乳腺癌细胞自噬

摘要/Abstract

摘要： 目的探讨丝胶对PI3K/Akt/mTOR信号通路的调控及其对三阴性乳腺癌（TNBC) MDA-MB-468细胞自噬的诱导作用,并阐明作用靶点。方法将MDA-MB-468细胞随机分为对照组（不含药物）、实验组（8 mg·mL^-1丝胶）、Akt激活剂组（8 mg·mL^-1丝胶+10 μmol·L^-1SC79）,分别给予相应药物作用24 h后,采用RT-qPCR法检测各组细胞PI3K、Akt、mTOR、自噬效应蛋白（Beclin1）、自噬相关蛋白5（ATG5）、微管相关轻链蛋白3（LC3）、螯合体1（P62）mRNA表达水平;Western blotting法检测各组细胞PI3K、Akt、p-Akt、mTOR、p-mTOR、Beclin1、ATG5、LC3、P62蛋白表达水平;免疫荧光染色法检测各组细胞LC3、P62蛋白的定位及荧光强度。结果与对照组相比,实验组MDA-MB-468细胞PI3K、Akt、mTOR、P62 mRNA及PI3K、Akt、p-Akt、mTOR、p-mTOR、P62蛋白水平均显著降低（P<0.05）,Beclin1、ATG5、LC3 mRNA水平及Beclin1、ATG5、LC3Ⅱ/LC3Ⅰ蛋白水平均显著升高（P<0.05）;与实验组相比,Akt激活剂组MDA-MB-468细胞PI3K、Akt、mTOR、P62 mRNA及PI3K、Akt、p-Akt、mTOR、p-mTOR、P62蛋白水平均显著升高（P<0.05）,Beclin1、ATG5、LC3 mRNA水平及Beclin1、ATG5、LC3Ⅱ/LC3Ⅰ蛋白水平均显著降低（P<0.05）。结论丝胶对MDA-MB-468细胞自噬的诱导作用可通过以Akt为靶点调控PI3K/Akt/mTOR信号通路的转导来实现。

关键词: 丝胶, 三阴性乳腺癌, PI3K/Akt/mTOR信号通路, 自噬, MDA-MB-468细胞

Abstract: Objective To investigate the induction of autophagy of MDA-MB-468 cells in triple-negative breast cancer (TNBC) by sericin through regulating the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway, and to clarify the target of action. Methods MDA-MB-468 cells were randomly divided into a control group (without drugs), an experimental group (8 mg·mL^-1 sericin), and an Akt activator group (8 mg·mL^-1 sericin + 10 μmol·L^-1SC79). After 24 hours treatment with corresponding drugs, the mRNA expression levels of PI3K, Akt, mTOR, autophagy effector protein (Beclin1), autophagy associated protein 5 (ATG5), microtubule associated light chain protein3 (LC3) and chelator 1 (P62) in cells of each group were detected by RT qPCR. The protein expression levels of PI3K, Akt, p-Akt, mTOR, p-mTOR, Beclin1, ATG5, LC3 and P62 in cells of each group were detected by Western blotting. The localization and fluorescence intensity of LC3 and P62 proteins in cells of each group were detected by immunofluorescence staining. Results Compared with the control group, the mRNA levels of PI3K, Akt, mTOR, and P62 and the protein levels of PI3K, Akt, p-Akt, mTOR, p-mTOR, and P62 in MDA-MB-468 cells of the experimental group were significantly decreased (P<0.05), while the mRNA levels of Beclin1, ATG5, LC3 and the protein levels of Beclin1, ATG5, LC3Ⅱ/LC3Ⅰ were significantly increased (P<0.05). Compared with the experimental group, the mRNA levels of PI3K, Akt, mTOR, P62 and the protein levels of PI3K, Akt, p-Akt, mTOR, p-mTOR, P62 in MDA-MB-468 cells of the Akt activator group were significantly increased (P<0.05), while the mRNA levels of Beclin1, ATG5, LC3 and the protein levels of Beclin1, ATG5, LC3Ⅱ/LC3Ⅰ were significantly decreased (P<0.05). Conclusion The induction effect of sericin on autophagy of MDA-MB-468 cells can be achieved by targeting Akt to regulate the transduction of the PI3K/Akt/mTOR signaling pathway.

Key words: sericin, triple-negative breast cancer, PI3K/Akt/mTOR signaling pathway, autophagy, MDA-MB-468 cells

中图分类号:

R587.1

李警耀, 张睿, 金美琪, 陈志宏. 丝胶靶向Akt调控PI3K/Akt/mTOR信号通路诱导三阴乳腺癌细胞自噬[J]. 承德医学院学报, 2025, 42(6): 456-462.

LI Jingyao, ZHANG Rui, JIN Meiqi, CHEN Zhihong. Sericin targets akt to regulate the PI3K/Akt/mTOR signaling pathway to induce autophagy in triple-negative breast cancer cells[J]. Journal of Chengde Medical University, 2025, 42(6): 456-462.

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链接本文: http://tougao.cdmc.edu.cn:8088/CN/

http://tougao.cdmc.edu.cn:8088/CN/Y2025/V42/I6/456

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