二甲双胍调控AKT/mTOR通路抑制前列腺癌的机制

Metformin Inhibits Prostate Cancer Progression via Regulation of the AKT/mTOR Signaling Pathway

摘要

目的 探讨二甲双胍对前列腺癌细胞增殖、迁移和侵袭的影响，并分析其是否通过调控人类抗原R（human antigen R, HuR）及AKT/mTOR信号通路发挥抗肿瘤作用。方法 取人前列腺癌细胞PC3和22RV1细胞系，采用二甲双胍干预，并结合HuR敲低和过表达实验进一步验证其作用机制。采用细胞计数试剂盒-8（cell counting kit-8, CCK-8）检测细胞增殖活性，Transwell实验检测细胞迁移和侵袭能力，实时荧光定量PCR（quantitative real-time polymerase chain reaction, qRT-PCR）和Western blot检测HuR及AKT/mTOR信号通路相关蛋白的表达水平。结果 二甲双胍干预可降低PC3和22RV1细胞的增殖、迁移和侵袭能力，并抑制AKT/mTOR通路相关蛋白的表达（P<0.05）。敲低HuR后，前列腺癌细胞的增殖、迁移和侵袭能力同样下降，AKT/mTOR通路活性受到抑制。相反，过表达HuR可部分减弱二甲双胍对前列腺癌细胞恶性生物学行为及AKT/mTOR通路的抑制作用。此外，二甲双胍可呈浓度依赖性下调PC3和22RV1细胞中HuR的mRNA和蛋白表达水平（P<0.01）。结论 二甲双胍可能通过下调HuR表达，抑制AKT/mTOR信号通路活化，从而降低前列腺癌细胞的增殖、迁移和侵袭能力。

Abstract

Objective To investigate the effects of metformin on the proliferation, migration, and invasion of prostate cancer cells, and to determine whether its antitumor effects are mediated by regulation of human antigen R (HuR) and the AKT/mammalian target of rapamycin (mTOR) signaling pathway. Methods Human prostate cancer cell lines PC3 and 22RV1 were used as the research models and were treated with metformin. HuR knockdown and overexpression experiments were performed to further verify the underlying mechanism. Cell proliferation was assessed using the cell counting kit-8 (CCK-8) assay. Cell migration and invasion were evaluated by Transwell assays. The expression levels of HuR and AKT/mTOR pathway-related proteins were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. Results Metformin treatment reduced the proliferation, migration, and invasion abilities of PC3 and 22RV1 cells, and inhibited the expression of proteins associated with the AKT/mTOR signaling pathway (P < 0.05). HuR knockdown also decreased the proliferation, migration, and invasion abilities of prostate cancer cells and inhibited AKT/mTOR pathway activity. In contrast, HuR overexpression partially attenuated the inhibitory effects of metformin on the malignant biological behaviors of prostate cancer cells and its inhibitory effects on the AKT/mTOR signaling pathway. In addition, metformin downregulated HuR mRNA and protein expression levels in PC3 and 22RV1 cells in a concentration-dependent manner ( P < 0.01). Conclusion Metformin may inhibit the activation of the AKT/mTOR signaling pathway by downregulating HuR expression, thereby reducing the proliferation, migration, and invasion of prostate cancer cells.

关键词

前列腺癌 / 二甲双胍 / 人类抗原R / AKT / mTOR

Key words

Prostate cancer / Metformin / Human antigen R / AKT / mTOR

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