Background

Sphingosine kinase 1 (Sphk1) is abnormally expressed in various tumors. This study explored the effects of Sphk1 in the polarity of breast cancer (BC) epithelial cells and on stem cell characteristicst.

Materials & methods

Reverse transcription quantitative PCR (RT-qPCR) was performed to detect Sphk1 levels in human mammary epithelial cells (MCF-10A) and BC cell lines (MCF-7, T47D, SKBR3, MDA-MB-231, and BT-474). After Sphk1-overexpression or Sphk1 silencing, the morphology of cells and stem cell-like properties of BC cells were analyzed. Metastasis of BC cells was assessed by wound healing and Transwell assays. Western blotting was performed to detect levels of epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, N-cadherin and Vimentin) and stem cell-specific markers (SOX2, OCT4, NANOG and ALDH1).

Results

Sphk1 was increased in BC cell lines than MCF-10A. Sphk1 induced EMT, regulated expression of EMT-related proteins, and accelerated the migration and invasion of BC cells. Silencing Sphk1 inhibited the sphere formation and down-regulated the expression of stem cell-specific markers, whereas Sphk1-overexpression contributed to the maintenance of the characteristics of mammary stem cells.

Conclusion

Sphk1 induces migration in BC cells and promotes stem cell characteristics by regulating EMT, The current findings provide a new potential for developing targeted therapy for tumor treatment

中文翻译：

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Sphk1通过调节上皮-间质转化和干细胞特性参与乳腺癌的恶性进展。

背景

鞘氨醇激酶1（Sphk1）在各种肿瘤中异常表达。这项研究探讨了Sphk1对乳腺癌（BC）上皮细胞极性和干细胞特性的影响。

材料与方法

进行逆转录定量PCR（RT-qPCR）以检测人乳腺上皮细胞（MCF-10A）和BC细胞系（MCF-7，T47D，SKBR3，MDA-MB-231和BT-474）中Sphk1的水平。Sphk1过表达或Sphk1沉默后，分析细胞的形态和BC细胞的干细胞样性质。通过伤口愈合和Transwell测定法评估BC细胞的转移。进行了蛋白质印迹，以检测上皮-间质转化（EMT）相关蛋白（E-钙黏着蛋白，N-钙黏着蛋白和波形蛋白）和干细胞特异性标记（SOX2，OCT4，NANOG和ALDH1）的水平。

结果

Sphk1在BC细胞系中比MCF-10A增加。Sphk1诱导EMT，调节EMT相关蛋白的表达，并加速BC细胞的迁移和侵袭。沉默Sphk1抑制了球的形成并下调了干细胞特异性标志物的表达，而Sphk1的过表达则有助于维持乳腺干细胞的特性。

结论

Sphk1通过调节EMT诱导BC细胞迁移并促进干细胞特性，目前的发现为开发靶向治疗肿瘤的方法提供了新的潜力