鞘氨醇激酶 1 通过激活 JAK/STAT 通路和上调结肠癌细胞中 S1PR1 的表达促进癌症进展。

SphK1 Promotes Cancer Progression through Activating JAK/STAT Pathway and Up-Regulating S1PR1 Expression in Colon Cancer Cells.

机构信息

Department of Medicinal Oncology, The First Affiliated Hospital, SUN Yat-Sen University, Guangzhou, 510080, China.

Department of Clinical Nutrition, The First Affiliated Hospital, SUN Yat-Sen University, Guangzhou, 510080, China.

出版信息

Anticancer Agents Med Chem. 2022;22(2):254-260. doi: 10.2174/1871520621666210401105344.

DOI:10.2174/1871520621666210401105344

PMID:33797381

Abstract

BACKGROUND

SphK1 is a conserved lipid kinase, which can catalyze the formation of tumorpromoting factor sphingosine phosphate-1 (S1P).

OBJECTIVE

This study aimed to investigate the effect of SphK1 on the proliferation/migration of colon cancer cells and associated mechanisms.

METHODS

Transcription of the SphK1 gene in colon cancer cells was detected. Gene transcription of SphK1 was inhibited by transfecting with the si-SphK1 gene in colon cancer cells. Effects of SphK1 inhibition (si-SphK1) on cell migration/proliferation were detected using the transwell system and MTS. Gene transcription of SIP, S1PR1, S1PR2, S1PR3, and activation of JAK/STAT3 pathway were examined using RT-PCR and western blot assay. S1PR1 over-expressing plasmid was constructed and transfected into cells. Effects of S1PR1 overexpression on migration/proliferation of si-SphK1 transfected colon cancer cells and activation of JAK/STAT3 pathway were determined using RT-PCR and western blotting.

RESULTS

Gene transcription of SphK1 in SW480 and HT-29 colon cancer cells was significantly inhibited by transfection of the si-SphK1 gene. Transwell migration and MTS findings showed that si-SphK1 transfection (si- SphK1 group) could reduce migration quantity and cell viability of colon cancer cells compared to the negative control (NC) (p<0.0001). SphK1 inhibition (si-SphK1 group) significantly down-regulated S1PR1 and S1PR3 gene transcription in SW480 and HT-29 cells (p<0.0001) and decreased activation level of JAKSTAT3 signaling pathway compared to NC group (p<0.05). Over-expression of S1PR1 reversed inhibitory effects of si-SphK1 on migration/proliferation of SW480 and activation of JAK/Stat3.

CONCLUSION

SphK1 promoted proliferation and migration of colon cancer cells through promoting JAK/STAT activation and up-regulating S1PR1 expression.

摘要

背景

SphK1 是一种保守的脂质激酶，可催化肿瘤促进因子神经鞘氨醇磷酸-1（S1P）的形成。

目的

本研究旨在探讨 SphK1 对结肠癌细胞增殖/迁移的影响及其相关机制。

方法

检测结肠癌细胞中 SphK1 基因的转录。通过转染 si-SphK1 基因抑制结肠癌细胞中 SphK1 的基因转录。用 Transwell 系统和 MTS 检测 SphK1 抑制（si-SphK1）对细胞迁移/增殖的影响。用 RT-PCR 和 Western blot 检测 SIP、S1PR1、S1PR2、S1PR3 的基因转录和 JAK/STAT3 通路的激活。构建 S1PR1 过表达质粒并转染细胞。用 RT-PCR 和 Western blot 检测 S1PR1 过表达对 si-SphK1 转染的结肠癌细胞迁移/增殖和 JAK/STAT3 通路激活的影响。

结果

SW480 和 HT-29 结肠癌细胞中 SphK1 的基因转录经 si-SphK1 基因转染后明显受到抑制。Transwell 迁移和 MTS 结果显示，与阴性对照（NC）相比，si-SphK1 转染（si-SphK1 组）可减少结肠癌细胞的迁移数量和细胞活力（p<0.0001）。SphK1 抑制（si-SphK1 组）显著下调 SW480 和 HT-29 细胞中 S1PR1 和 S1PR3 的基因转录（p<0.0001），并降低 JAK-STAT3 信号通路的激活水平与 NC 组相比（p<0.05）。S1PR1 的过表达逆转了 si-SphK1 对 SW480 细胞迁移/增殖和 JAK/Stat3 激活的抑制作用。