SK1 Inhibitor RB005 Induces Apoptosis in Colorectal Cancer Cells through SK1 Inhibition Dependent and Independent Pathway.

IF 2.4 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Jitendra Shrestha, Maftuna Shamshiddinova, Yong-Moon Lee, Yoon Sin Oh, Dong Jae Baek, Eun-Young Park
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引用次数: 3

Abstract

Background and objective: Colorectal cancer (CRC) is the fourth leading cause of cancer- related death globally, with a high incidence rate in economically fast-growing countries. Sphingosine- 1-phosphate (S1P) is a bioactive lipid mediator that plays critical roles in cancer cell proliferation, migration, and angiogenesis converted by the isoforms of sphingosine kinase (SK1 and SK2). SK1 is highly expressed in colorectal cancer; its inhibitors suppress the formation of S1P and increase ceramide levels having a pro-apoptotic function. RB005 is a selective SK1 inhibitor and a structural analog of PP2A activator FTY720. The purpose of this study is to test whether RB005, an SK1 inhibitor, can be used as an anticancer agent by inhibiting the growth of colon cancer cells.

Methods: We performed MTT and colony-forming assay using colon cancer cell lines HT29 and HCT116 cells to examine the cell toxicity effect of RB005. To determine whether apoptosis of RB005 in colon cancer cell line is due to SK1 inhibition or other mechanisms due to its structural similarity with FTY720, we conducted LC/MS, siRNA knockdown, and PP2A activity experiments.

Results: RB005 notably inhibited CRC cell growth and proliferation compared to PF543 and ABC294640 by inducing the mitochondria-mediated intrinsic apoptotic pathway. Apoptotic cell death is caused by increased mitochondrial permeability Initiated by the activation of pro-apoptotic protein BAX, increased ceramides, and activation of PP2A. Also, RB005 treatment in HT29 cells did not change the expression level of SK1, but strikingly decreased SK1 activity and S1P levels. All these events of cell death and apoptosis were less effective when SK1 was knocked down by siRNA.

Conclusion: This result indicates that RB005 shows the in-vitro anti-CRC effect by inhibiting SK1 activity and PP2A activation, increasing proapoptotic ceramide levels following the activation of the intrinsic apoptotic pathway.

SK1抑制剂RB005通过SK1抑制依赖和独立通路诱导结直肠癌细胞凋亡
背景与目的:结直肠癌(CRC)是全球癌症相关死亡的第四大原因,在经济快速增长的国家发病率很高。鞘氨醇- 1-磷酸(S1P)是一种生物活性脂质介质,在鞘氨醇激酶(SK1和SK2)亚型转化的癌细胞增殖、迁移和血管生成中起关键作用。SK1在结直肠癌中高表达;其抑制剂抑制S1P的形成并增加具有促凋亡功能的神经酰胺水平。RB005是一种选择性SK1抑制剂,是PP2A活化剂FTY720的结构类似物。本研究的目的是测试SK1抑制剂RB005是否可以通过抑制结肠癌细胞的生长作为抗癌药物。方法:采用结肠癌细胞系HT29和HCT116细胞进行MTT和集落形成实验,观察RB005的细胞毒性作用。为了确定结肠癌细胞系中RB005的凋亡是由于其与FTY720的结构相似性导致SK1抑制还是其他机制,我们进行了LC/MS、siRNA敲低和PP2A活性实验。结果:与PF543和ABC294640相比,RB005通过诱导线粒体介导的内在凋亡途径显著抑制结直肠癌细胞的生长和增殖。凋亡细胞死亡是由促凋亡蛋白BAX的激活、神经酰胺的增加和PP2A的激活引发的线粒体通透性增加引起的。RB005在HT29细胞中的作用也没有改变SK1的表达水平,但显著降低了SK1活性和S1P水平。当SK1被siRNA敲低时,所有这些细胞死亡和凋亡事件都不那么有效。结论:RB005通过抑制SK1活性和PP2A激活,增加促凋亡神经酰胺水平,激活细胞固有凋亡通路,具有体外抗crc作用。
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来源期刊
Current molecular pharmacology
Current molecular pharmacology Pharmacology, Toxicology and Pharmaceutics-Drug Discovery
CiteScore
4.90
自引率
3.70%
发文量
112
期刊介绍: Current Molecular Pharmacology aims to publish the latest developments in cellular and molecular pharmacology with a major emphasis on the mechanism of action of novel drugs under development, innovative pharmacological technologies, cell signaling, transduction pathway analysis, genomics, proteomics, and metabonomics applications to drug action. An additional focus will be the way in which normal biological function is illuminated by knowledge of the action of drugs at the cellular and molecular level. The journal publishes full-length/mini reviews, original research articles and thematic issues on molecular pharmacology. Current Molecular Pharmacology is an essential journal for every scientist who is involved in drug design and discovery, target identification, target validation, preclinical and clinical development of drugs therapeutically useful in human disease.
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