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.
Anticancer Agents Med Chem. 2022;22(2):254-260. doi: 10.2174/1871520621666210401105344.
SphK1 is a conserved lipid kinase, which can catalyze the formation of tumorpromoting factor sphingosine phosphate-1 (S1P).
This study aimed to investigate the effect of SphK1 on the proliferation/migration of colon cancer cells and associated mechanisms.
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.
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.
SphK1 promoted proliferation and migration of colon cancer cells through promoting JAK/STAT activation and up-regulating S1PR1 expression.
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。
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。
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。
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。
SphK1 通过促进 JAK/STAT 激活和上调 S1PR1 表达促进结肠癌细胞的增殖和迁移。
