可利霉素体外抗结肠癌活性的研究  

Study on the activities of carrimycin on colon cancer in vitro

作　　者：刘娟娟[1,2] 戴剑漉[1] 高晓杰 赫卫清[1,2] LIU Juan-juan;DAI Jian-lu;GAO Xiao-jie;HE Wei-qing(NHC Key Laboratory of Biotechnology of Antibiotics,Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences and Peking Union Medical College,Bejing 100050,China;CAMS Key Laboratory of Synthetic Biology for Drug Innovation,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所卫健委抗生素生物工程重点实验室,北京100050 [2]中国医学科学院北京协和医学院、药物合成生物学重点实验室,北京100050

出　　处：《药物分析杂志》2022年第7期1142-1152,共11页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金项目(82104233,82073900)。

摘　　要：目的:评价可利霉素(carrimycin, CAM)及其单组分异戊酰螺旋霉素Ⅰ(isovalerylspiramycinⅠ, ISPⅠ)对结肠癌细胞的抑制作用,初步探讨其抗肿瘤的作用机制。方法:选择结肠癌细胞系LoVo,评价CAM及其单组分ISPⅠ与螺旋霉素(spiramycin, SP)及其单组分螺旋霉素Ⅰ(spiramycinⅠ, SPⅠ)的抗肿瘤活性,采用CCK-8试剂盒检测对细胞增殖的影响;采用克隆形成实验分析对细胞克隆形成能力的影响;采用流式细胞术分析对细胞凋亡的影响;采用划痕实验检测对细胞迁移能力的影响。通过转录组分析CAM、ISPⅠ发挥抗肿瘤活性可能作用的信号通路。结果:CAM及其单组分ISPⅠ可以有效抑制结肠癌细胞的增殖、克隆形成,能促使细胞凋亡比例增加,但对细胞迁移能力影响不明显;在给药6 h和9 h后,CAM及其单组分ISPⅠ在细胞内的相对含量高于SP及其单组分SPⅠ;转录组测序分析发现CAM及ISPⅠ可能作用于细胞粘附相关信号通路和PI3K/Akt信号通路。结论:CAM及其单组分ISPⅠ具有体外抗结肠癌活性,且优于SP及其单组分SPⅠ,这可能与其更好的细胞穿透性相关,CAM及ISPⅠ的抗结肠癌活性可能是通过作用于细胞粘附相关信号通路和PI3K/Akt信号通路而发挥的,这为进一步研究CAM及其单组分的抗肿瘤作用机制奠定了基础。Objective:To evaluate the inhibitory effect of carrimycin(CAM) and its single component, isovalerylspiramycin Ⅰ(ISP Ⅰ), and preliminarily discuss the mechanism of their antitumor effect. Methods: Colon cancer cell lines LoVo was selected to evaluate the antitumor activity of CAM, ISP Ⅰ, spiramycin(SP) and its single component spiramycin Ⅰ(SP Ⅰ). The effect on cell proliferation was detected by Cell Counting Kit-8. Cell colony formation ability was analyzed by clone formation assay. The effect on cell apoptosis was analyzed by flow cytometry. The effect on cell migration capacity was detected by scratch assay. Through transcriptome analysis, the possible signal pathways of CAM and ISP Ⅰ to exert antitumor activity were found. Results: CAM and itssingle component ISPⅠ were effective against the proliferation and colony formation of colon cancer cells, and increased the apoptosis proportion of LoVo cells.However, the effect of CAM and ISP Ⅰ on cell migration ability was not significant.The relative content of CAM and its single component ISP Ⅰ were all higher than that of SP and SP Ⅰ after 6 h and 9 h of administration. Transcriptome sequencing analysis showed that CAM and ISP Ⅰ might act on cell adhesion related and PI3 K/Akt signaling pathways. Conclusion: The anti-colonic cancer activities of CAM and its single component ISP Ⅰ were superior to that of SP and its single component SP Ⅰ in vitro, which one of reasons was probably attributed to their better cell penetration ability. The anti-colonic cancer activities of CAM and ISP Ⅰ may be induced by their effects on cell adhesion related signaling pathway and PI3 K/Akt signaling pathway, which lays a foundation for further study on antitumor mechanism of CAM and its single component.

关 键 词：可利霉素 异戊酰螺旋霉素Ⅰ 螺旋霉素 结肠癌LOVO细胞 增殖 凋亡 迁移 转录组 

分 类 号：R917[医药卫生—药物分析学]