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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (6): 633-636.doi: 10.12047/j.cjap.6026.2020.132

• 研究论文 • 上一篇    下一篇

川楝素对人胃癌细胞MKN-45中CTPS细胞蛇形成的影响及其机制*

陈雯1, 张伟伟1,2△, 潘阳1, 刘畅1, 邵淑丽1,2   

  1. 1.齐齐哈尔大学生命科学与农林学院, 齐齐哈尔 161006;
    2.抗性基因工程与寒地生物多样性保护黑龙江省重点实验室, 齐齐哈尔 161006
  • 收稿日期:2020-03-06 修回日期:2020-11-25 出版日期:2020-11-28 发布日期:2021-03-15
  • 通讯作者: Tel: 0452-2738219; E-mail: zww121@163.com
  • 基金资助:
    *国家青年科学基金项目(31801148);黑龙江省自然科学基金项目(C2016057);黑龙江省教育厅基本业务专项(135209260);黑龙江省省属高等学校基本科研业务费科研项目(植物性食品加工技术特色学科专项)(YSTSXK201874)

Effects of Toosendanin on the formation of CTPS cytoophidium in human gastric cancer cell MKN-45 and its mechanism

CHEN Wen1, ZHANG Wei-wei1,2△, PAN Yang1, LIU Chang1, SHAO Shu-li1,2   

  1. 1. College of Life Sciences, Agriculture and Forestry, Qiqihar161006, China;
    2. Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas, Qiqihar University, Qiqihar161006, China
  • Received:2020-03-06 Revised:2020-11-25 Online:2020-11-28 Published:2021-03-15

摘要: 目的:本研究旨在对川楝素(TSN)与胃癌细胞MKN-45中CTPS细胞蛇形成的联系进行初步探究。方法:以人胃癌细胞MKN-45为实验材料,设置7个处理组分别为:0、20、40、60、80、100、120 nmol/L TSN。每组3次重复,分别作用24 h、48 h、72 h,利用CCK-8法检测川楝素对MKN-45细胞增殖抑制作用,使用免疫荧光检测之后再通过激光共聚焦显微镜观察细胞内CTPS细胞蛇形态,qRT-PCR检测川楝素对MYC基因表达的影响。另外设置2个处理组为1 mmol/L DON和1 mmol/L MPA,每组3次重复,作用6 h然后采用免疫荧光检测细胞蛇形态。结果:免疫荧光结果显示,分别利用1 mmol/L DON和1 mmol/L MPA 处理MKN-45细胞后CTPS形成丝状的细胞蛇结构,意味着该细胞具有形成CTPS细胞蛇的能力;川楝素处理组的细胞增殖率明显低于0 nmol/L TSN组(P< 0.01);免疫荧光结果显示80 nmol/L的川楝素作用72 h时MKN-45细胞中CTPS细胞蛇形成数量最多;qRT-PCR检测结果显示,80 nmol/L川楝素作用24 h细胞内MYC表达明显降低(P<0.05),48 h后细胞内MYC的表达量明显增多(P<0.01)随后表达降低。结论:川楝素可能通过调节MYC的表达影响细胞内细胞蛇的组装。

关键词: 川楝素, CTPS细胞蛇, MYC, MKN-45细胞

Abstract: Objective: To investigate the relationship between toosendanin(TSN) and CTP synthase(CTPS) cytoophidium formation in gastric cancer MKN-45 cells. Methods: In this study, the experimental material is MKN-45 human gastric cancer cells. It contains 7 treatment groups of 0, 20, 40, 60, 80, 100, and 120 nmol/L TSN. Each group was treated in triplex privately for 24、48 and 72 hours. Cell counting kit-8 (CCK8) was used to detect the inhibitory effect of TSN on the proliferation of MKN-45 cells. After immunofluorescence detection, the morphology of CTPS cells was observed by a laser confocal microscope. The effect of TSN on MYC gene expression was detected by qRT-PCR. In addition, it contains 2 treatment groups of 1 mmol/L DON and 1 mmol/L MPA, each group was treated in triplex privately for 6 hours and then the cytoophidium morphology was detected by immunofluorescence. Results: The results of immunofluorescence showed that CTPS formed a filamentous cytoophidium structure after treating MKN-45 cells with 1 mmol/L DON and 1 mmol/L MPA, which means that the cells have the ability to form CTPS cytoophidium; The cell proliferation rate of TSN treatment group was significantly lower than that of 0 nmol / L TSN group (P<0.01); Immunofluorescence results showed that CTPS cytoophidium was the most abundant in MKN-45 cells after treated with 80 nmol/L TSN for 72 h. The results of qRT-PCR showed that MYC expression in cells was significantly decreased after treated with 80 nmol/L TSN for 24 h (P<0.05), and MYC expression was significantly increased after 48 h (P<0.01), and then decreased. Conclusion: Toosendanin may affect intracellular cytoophidium assembling by regulating the expression of MYC.

Key words: Toosendanin, CTPS cytoophidium, MYC, MKN-45 cell

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