Purpose

Purine metabolism involves various intracellular and extracellular enzymes, including cN-II and CD73 that dephosphorylate intracellular and extracellular nucleoside monophosphates into their corresponding nucleosides. We conducted a study to better understand the biological roles of these enzymes in breast and lung cancer cells.

Methods

We modified cN-II and/or CD73 expression in human breast cancer cells (MDA-MB-231), human lung cancer cells (NCI-H292) and murine breast cancer cells (4T1) using the CRISPR/Cas9 technique, and evaluated their impact on various cellular parameters such as proliferation, migration, invasion, intracellular nucleotide pools and nucleotide metabolism-related gene expression under extracellular nucleotide stress conditions.

Results

Intracellular nucleotide contents were found to be altered in the modified cancer cell models both at their basal levels and after exposure to adenosine or AMP. Altered cN-II and CD73 levels were also found to be associated with cell migration and invasion alterations, involving TIMP-2, MMP-2 and MMP-9 expression, as well as alterations in the COX-2/PGE2/AKT pathway.

Conclusion

Our results highlight new cell-specific roles of cN-II and CD73 in cancer cell biology and provide insight into their interactions with different intracellular pathways.

中文翻译：

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通过 COX-2/PGE2/AKT 轴调节增强缺乏 cN-II 和 CD73 的乳腺癌和肺癌细胞的迁移。

目的

嘌呤代谢涉及各种细胞内和细胞外酶，包括 cN-II 和 CD73，它们将细胞内和细胞外的单磷酸核苷去磷酸化为相应的核苷。我们进行了一项研究，以更好地了解这些酶在乳腺癌和肺癌细胞中的生物学作用。

方法

我们使用 CRISPR/Cas9 技术修改了人乳腺癌细胞 (MDA-MB-231)、人肺癌细胞 (NCI-H292) 和鼠乳腺癌细胞 (4T1) 中的 cN-II 和/或 CD73 表达，并评估了它们的在细胞外核苷酸应激条件下，对各种细胞参数的影响，如增殖、迁移、侵袭、细胞内核苷酸库和核苷酸代谢相关基因表达。

结果

发现在修饰的癌细胞模型中，细胞内核苷酸含量在其基础水平和暴露于腺苷或 AMP 后都发生了改变。还发现改变的 cN-II 和 CD73 水平与细胞迁移和侵袭改变有关，包括 TIMP-2、MMP-2 和 MMP-9 表达，以及 COX-2/PGE2/AKT 通路的改变。

结论

我们的结果突出了 cN-II 和 CD73 在癌细胞生物学中的新细胞特异性作用，并提供了对它们与不同细胞内途径相互作用的深入了解。