Department of Orthopaedics, Affiliated Hospital of Jining Medical University, No. 129 Hehua Road, Jining, Shandong, China.
Department of Laboratory Medicine, Affiliated Hospital of Jining Medical University, No. 89 Guhuai Road, Jining, Shandong, China.
Clin Transl Oncol. 2022 Jul;24(7):1403-1412. doi: 10.1007/s12094-022-02786-y. Epub 2022 Feb 8.
The role of autophagy in prostate cancer metastasis remains controversial, and the effects of the autophagy-related gene ATG5 on prostate cancer metastasis are poorly understood. This study aims to explore the effects of ATG5 on prostate cancer metastasis and its molecular mechanism.
The metastatic characteristics of LNCaP and DU145 cells were assessed by NOD/SCID mouse experiments, western blot, transwell assay, and wound-healing assay. Double membrane autophagic vesicle observation and the adenovirus-expressing mCherry-GFP-LC3B fusion protein were used to assess the autophagic characteristics of LNCaP and DU145 cells. The role of p62 in the accumulation of TWIST1 was confirmed by western blot under different conditions. The lentivirus particles of shATG5, NOD/SCID mice experiments, western blot, transwell assay, and wound-healing assay were used to confirm the role of ATG5 in TWIST1 accumulation and prostate cancer cell metastasis.
We identified a stabilizing effect of p62 on TWIST1 in the autophagic regulation of EMT and prostate cancer metastasis. The loss of ATG5 in DU145 cells resulted in autophagy deficiency and p62 accumulation, which stabilized TWIST1 and increased the TWIST1 level in prostate cancer cells, and eventually promoted EMT and metastasis. In comparison, LNCaP cells with regular ATG5 expression and autophagy status retained remarkable epithelial cell characteristics and had limited metastatic characteristics. Similar results were also found in wild-type LNCaP cells and LNCaP cells with stable ATG5 interference.
Our research revealed ATG5-mediated autophagy as a key mechanism that controls the metastasis of prostate cancer by regulating p62 abundance and TWIST1 stabilization.
自噬在前列腺癌转移中的作用仍存在争议，并且自噬相关基因 ATG5 对前列腺癌转移的影响知之甚少。本研究旨在探讨 ATG5 对前列腺癌转移的影响及其分子机制。
通过 NOD/SCID 小鼠实验、Western blot、Transwell 检测和划痕愈合实验评估 LNCaP 和 DU145 细胞的转移特征。使用双膜自噬囊泡观察和表达 mCherry-GFP-LC3B 融合蛋白的腺病毒评估 LNCaP 和 DU145 细胞的自噬特征。通过 Western blot 在不同条件下证实 p62 在 TWIST1 积累中的作用。使用 shATG5、NOD/SCID 小鼠实验、Western blot、Transwell 检测和划痕愈合实验证实 ATG5 在 TWIST1 积累和前列腺癌细胞转移中的作用。
我们发现 p62 在 EMT 和前列腺癌转移的自噬调节中对 TWIST1 具有稳定作用。DU145 细胞中 ATG5 的缺失导致自噬缺陷和 p62 积累，从而稳定 TWIST1 并增加前列腺癌细胞中的 TWIST1 水平，最终促进 EMT 和转移。相比之下，具有正常 ATG5 表达和自噬状态的 LNCaP 细胞保留了显著的上皮细胞特征，并且具有有限的转移特征。在野生型 LNCaP 细胞和稳定干扰 ATG5 的 LNCaP 细胞中也发现了类似的结果。
我们的研究揭示了 ATG5 介导的自噬通过调节 p62 丰度和 TWIST1 稳定来控制前列腺癌转移的关键机制。
