自噬抑制减轻甲状腺癌细胞迁移和侵袭。

Inhibition of autophagy mitigates cell migration and invasion in thyroid cancer.

机构信息

Department of Surgery, The University of Cincinnati, Cincinnati, OH; Department of Cancer Biology, Vontz Center for Molecular Studies, The University of Cincinnati, Cincinnati, OH.

Department of Surgery, The University of Cincinnati, Cincinnati, OH.

出版信息

Surgery. 2022 Jan;171(1):235-244. doi: 10.1016/j.surg.2021.08.024. Epub 2021 Sep 24.

DOI:10.1016/j.surg.2021.08.024

PMID:34565609

Abstract

BACKGROUND

Autophagy is a highly conserved process for maintaining cellular homeostasis. Upregulation of autophagy promotes metastasis by promoting the cancer stem cell state while also stimulating tumor cell migration and invasion. We hypothesized that autophagy upregulation would be critical for cancer stem cell maintenance as well as cellular migration and invasion in thyroid cancer.

METHODS

Validated papillary (MDA-T32, MDA-T68), follicular (FTC-133), and anaplastic (ATC-8505c) human thyroid cancer cell lines in culture were first assessed for autophagic capacity after bafilomycin clamping. Cancer stem cells were quantified by flow cytometry for aldehyde dehydrogenase and thyrosphere formation assay. Scratch migration and Matrigel invasion assays were performed in the presence of known autophagy inhibitors: Lys05, chloroquine, and FIP200siRNA.

RESULTS

Autophagy activity was observed across all cell lines. Thyrosphere formation, aldehyde dehydrogenase activity, and CD44 expression were reduced with inhibition of autophagy in MDA-T32, MDA-T68, FTC-133, and 8505c cells. Similarly, cell migration and invasion were attenuated: 42% (FIP200siRNA), 78% (Lys05), P < .001 in MDA-T32 cells; 54% (FIP200siRNA), 67% (Lys05), P < .001 in MDA-T68 cells; 73% (FIP200siRNA), 71% (Lys05), P < .001) in FTC-133 cells; and 60% (FIP200siRNA), 90% (Lys05), P < .001 in 8505c cells. Invasion assays demonstrated a 73%, 39%, 75%, and 65.1% reduction in the presence of Lys05 in T32, T68, FTC-133, and 8505c cells, respectively. We observed similar reductions in invasion with FIP200siRNA: 61%, 62%, 55%, and 81.4% in T32, T68, FTC-133, and 8505c cells.

CONCLUSION

Autophagy is upregulated across multiple thyroid cancer subtypes. In thyroid cancer cell lines, inhibition of autophagy attenuates cancer stem cell viability, cell migration, and invasion suggesting a role for autophagy in the progression of thyroid cancer. Greater understanding of autophagy regulation in thyroid cancer will aid in developing targeted therapeutics.

摘要

背景

自噬是维持细胞内稳态的一种高度保守的过程。自噬的上调通过促进癌症干细胞状态促进转移，同时刺激肿瘤细胞迁移和侵袭。我们假设自噬的上调对于癌症干细胞的维持以及甲状腺癌细胞的迁移和侵袭至关重要。

方法

在氟霉素夹闭后，首先评估培养中的经验证的乳头状（MDA-T32、MDA-T68）、滤泡状（FTC-133）和间变性（ATC-8505c）人甲状腺癌细胞系的自噬能力。通过醛脱氢酶和甲状腺球体形成测定法通过流式细胞术定量测定癌症干细胞。在已知的自噬抑制剂：Lys05、氯喹和 FIP200siRNA 的存在下进行划痕迁移和 Matrigel 侵袭测定。

结果

在所有细胞系中均观察到自噬活性。在 MDA-T32、MDA-T68、FTC-133 和 8505c 细胞中抑制自噬后，甲状腺球体形成、醛脱氢酶活性和 CD44 表达减少。同样，细胞迁移和侵袭也减弱：42%（FIP200siRNA），78%（Lys05），P <.001 在 MDA-T32 细胞中；54%（FIP200siRNA），67%（Lys05），P <.001 在 MDA-T68 细胞中；73%（FIP200siRNA），71%（Lys05），P <.001 在 FTC-133 细胞中；60%（FIP200siRNA），90%（Lys05），P <.001 在 8505c 细胞中。侵袭测定显示 Lys05 存在时 T32、T68、FTC-133 和 8505c 细胞中的侵袭分别减少了 73%、39%、75%和 65.1%。我们观察到用 FIP200siRNA 进行侵袭时也有类似的减少：61%、62%、55%和 81.4%在 T32、T68、FTC-133 和 8505c 细胞中。