Spinosyn A and Its Derivative Inhibit Colorectal Cancer Cell Growth via the EGFR Pathway
Spinosyn A and Its Derivative Inhibit Colorectal Cancer Cell Growth via the EGFR Pathway
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DOI：
10.1021/acs.jnatprod.3c00276
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发表时间：
2023-09-08
影响因子：
5.1
通讯作者：
Luo，Zhi-Yong
中科院分区：
文献类型：
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作者：
Peng，Kunjian;Zou，Zizheng;Luo，Zhi-Yong
Spinosyn A （SPA）来源于土壤微生物Saccharopolyspora spinosa及其衍生物LM2I，对多种癌细胞具有潜在的抑制作用。然而，SPA和LM2I在抑制人类结直肠癌细胞生长中的作用及其分子机制尚不完全清楚。采用3-(4,5-二甲基噻唑-2-基-)-2,5-二苯基溴化四唑（MTT）法和菌落形成法检测细胞活力。根据7种结直肠癌（CRC）细胞系中SPA和LM2I的ic50值，筛选敏感细胞系（HT29和SW480）和不敏感细胞系（SW620和RKO）。使用GSE2509和GSE10843数据集鉴定敏感和不敏感细胞系之间的69个差异表达基因（DEGs）。通过基因本体（GO）、京都基因与基因组百科全书（KEGG）富集分析和蛋白-蛋白相互作用（PPI）分析来阐明DEGs的分子机制。采用Western blot检测deg的hub基因，并采用CRISPR/Cas9系统进行验证。我们的数据表明，SPA及其衍生物LM2I在7种结直肠癌细胞系和结直肠癌异种移植肿瘤中具有显著的抗增殖活性。生物信息学分析表明表皮生长因子受体（epidermal growth factor receptor， EGFR）是DEGs的枢纽基因，与SPA和LM2I在结直肠癌细胞系中的抑制作用有关。本研究还发现，SPA和LM2I在体外和体内均抑制EGFR通路。
Spinosyn A (SPA), derived from a soil microorganism,Saccharopolyspora spinosa, and its derivative, LM2I, has potential inhibitory effects on a variety of cancer cells. However, the effects of SPA and LM2I in inhibiting the growth of human colorectal cancer cells and the molecular mechanisms underlying these effects are not fully understood. Cell viability was tested by using a 3-(4,5-dimethylthiazol-2-yl-)-2,5-diphenyltetrazolium bromide (MTT) assay and a colony formation assay. On the basis of the IC50values of SPA and LM2I in seven colorectal cancer (CRC) cell lines, sensitive (HT29 and SW480) and insensitive (SW620 and RKO) cell lines were screened. The GSE2509 and GSE10843 data sets were used to identify 69 differentially expressed genes (DEGs) between sensitive and insensitive cell lines. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, and protein–protein interactions (PPI) were performed to elucidate the molecular mechanisms of the DEGs. The hub gene of the DEGs was detected by Western blot analysis and verified using the CRISPR/Cas9 system. Our data indicate that SPA and its derivative LM2I have significant antiproliferative activity in seven colorectal cancer cell lines and colorectal xenograft tumors. On the basis of bioinformatics analysis, it was demonstrated that epidermal growth factor receptor (EGFR) was the hub gene of the DEGs and was associated with the inhibitory effects of SPA and LM2I in CRC cell lines. The study also revealed that SPA and LM2I inhibited the EGFR pathway in vitro and in vivo.