菊藻丸对乳腺癌MCF-7细胞PTEN/PI3K/Akt/mTOR信号通路的影响*

作者：欧阳志1，郑裴凡2，李松莲1

单位：1.湖南中医药大学第二附属医院，湖南 长沙 410005； 2.湖南中医药大学，湖南 长沙 410208

引用：引用：欧阳志，郑裴凡，李松莲.菊藻丸对乳腺癌MCF-7细胞PTEN/PI3K/Akt/mTOR信号通路的影响[J].中医药导报,2025,31(4):34-41.

DOI：10.13862/j.cn43-1446/r.2025.04.006

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摘要：

目的：探究菊藻丸含药血清通过调控PTEN/PI3K/Akt/mTOR信号通路抑制人乳腺癌MCF-7细胞恶性行为的作用及机制。方法：设计合成针对人PTEN基因的shRNA及阴性对照NC序列，构建PTEN基因沉默的慢病毒载体，分别感染MCF-7细胞。Real-time PCR和Western blotting验证PTEN敲减效果。将细胞分为正常对照组、空白血清组、菊藻丸含药血清组、LV-shNC组、LV-shNC+菊藻丸含药血清组、LV-shPTEN组、LV-shPTEN+菊藻丸含药血清组、LV-shPTEN+菊藻丸含药血清+MK2206组。CCK-8和集落形成实验检测细胞增殖能力，TUNEL染色检测凋亡水平，流式细胞术分析细胞周期分布，划痕实验和Transwell小室实验分别检测细胞迁移和侵袭能力，Western blotting检测PTEN/PI3K/Akt/mTOR通路关键蛋白表达变化。结果：Real-time PCR和Western blotting结果证实，PTEN shRNA转染MCF-7细胞后，PTEN mRNA和蛋白表达水平明显降低。与空白血清组比较，菊藻丸含药血清组MCF-7细胞增殖、集落形成、迁移侵袭能力显著下降，凋亡水平升高，G1期阻滞更明显，PTEN表达量升高，p-PI3K、p-AKT、p-mTOR水平降低（P<0.05）；LV-shNC组与空白血清组各指标比较，差异均无统计学意义（P>0.05）；LV-shNC+菊藻丸含药血清组较LV-shNC组细胞增殖、集落形成、迁移侵袭能力受到抑制，凋亡率及G1期比例升高，PTEN表达上调，PI3K/Akt/mTOR通路活性下降（P<0.05）；与LV-shNC+菊藻丸含药血清组比较，LV-shPTEN+菊藻丸含药血清组细胞增殖、侵袭转移能力增强，凋亡水平及G1期比例下降，PTEN表达量降低，PI3K/Akt/mTOR通路过度活化（P<0.05）；LV-shPTEN+菊藻丸含药血清+MK2206组较LV-shPTEN+菊藻丸含药血清组细胞恶性表型受到更明显抑制，PI3K/Akt/mTOR通路活性进一步降低（P<0.05）。结论：菊藻丸含药血清可通过激活PTEN，抑制PI3K/Akt/mTOR信号通路，进而抑制乳腺癌MCF-7细胞的增殖、侵袭转移，促进其凋亡和G1期阻滞。这可能是菊藻丸发挥抗乳腺癌作用的重要机制之一。

关键词：乳腺癌；菊藻丸；PTEN；PI3K/Akt/mTOR；细胞增殖；侵袭转移；凋亡；细胞周期

Abstract：

Objective: To investigate the effects and mechanisms of Juzao Pill-containing serum on inhibiting the malignant behaviors of human breast cancer MCF-7 cells through regulating the PTEN/PI3K/Akt/mTOR signaling pathway. Methods: shRNA targeting human PTEN gene and negative control NC sequences were designed and synthesized. Lentiviral vectors for PTEN gene silencing were constructed and used to infect MCF-7 cells. The knockdown effect of PTEN was verified by Real-time PCR and Western blotting. Cells were divided into control group, blank serum group, Juzao Pill-containing serum group, LV-shNC group, LV-shNC+Juzao Pill-containing serum group, LV-shPTEN group, LV-shPTEN+Juzao Pill-containing serum group, and LV-shPTEN+Juzao Pill-containing serum+MK2206 group. Cell proliferation was detected by CCK-8 and colony formation assays. Apoptosis level was examined by TUNEL staining. Cell cycle distribution was analyzed by flow cytometry. Scratch wound healing and Transwell assays were performed to assess cell migration and invasion abilities, respectively. Key protein expression changes in the PTEN/PI3K/Akt/mTOR pathway were detected by Western blotting. Results: Real-time PCR and Western blotting confirmed that PTEN mRNA and protein expression levels were significantly reduced after PTEN shRNA transfection in MCF-7 cells. Compared with the blank serum group, the Juzao Pill-containing serum group showed significantly decreased proliferation, colony formation, migration and invasion abilities, increased apoptosis level, more obvious G1 phase arrest, upregulated PTEN expression, and reduced phosphorylation levels of p-PI3K, p-Akt, and p-mTOR (P<0.05). There was no statistical difference in each indicator between the LV-shNC group and the blank serum group (P>0.05). Compared with the LV-shNC group, the LV-shNC+Juzao Pill-containing serum group exhibited inhibited cell proliferation, colony formation, migration and invasion abilities, increased apoptosis rate and G1 phase proportion, upregulated PTEN expression, and decreased PI3K/Akt/mTOR pathway activity (P<0.05). Compared with the LV-shNC+Juzao Pill-containing serum group, the LV-shPTEN+Juzao Pill-containing serum group showed enhanced cell proliferation, invasion and metastasis abilities, decreased apoptosis level and G1 phase proportion, reduced PTEN expression, and hyperactivated PI3K/Akt/mTOR pathway (P<0.05). The LV-shPTEN+Juzao Pill-containing serum+MK2206 group exhibited more obvious suppression of malignant phenotypes and further reduced PI3K/Akt/mTOR pathway activity compared with the LV-shPTEN+Juzao Pill-containing serum group (P<0.05). Conclusion: Juzao Pill-containing serum can inhibit the proliferation, invasion and metastasis, and promote the apoptosis and G1 phase arrest of breast cancer MCF-7 cells by activating PTEN and inhibiting the PI3K/Akt/mTOR signaling pathway, which may be one of the important mechanisms of Juzao Pill in exerting anti-breast cancer effects.

Key words：breast cancer; Juzao Pill; PTEN; PI3K/Akt/mTOR; cell proliferation; invasion and metastasis; apoptosis; cell cycle

发布时间：2025-12-20

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