青蒿琥酯通过调控Skp2、P21基因的表达抑制食管鳞癌细胞的研究  

Artesunate inhibits development of esophageal squamous cell carcinoma via regulating expression of Skp2 and P21

作　　者：张敏[1,2] 孙丽婷 沈冠彤 秦楠 胡晓玲 杨洁[5] 张永华[6] 师如意 ZHANG Min;SUN Li-ting;SHEN Guan-tong;QIN Nan;HU Xiao-ling;YANG Jie;ZHANG Yong-hua;SHI Ru-yi(Dept of Medical Cell Biology and Genetics,Shanxi Medical University,Taiyuan 030001,China;Translational Medicine Research Center,Shanxi Medical University,Taiyuan 030001,China;College of Pharmaceutical and Food Engineering,Shanxi University of Chinese Medicine,Jinzhong 030619,China;Dept of Pharmacology,Shanxi Medical University,Shanxi Medical University,Taiyuan 030001,China;Dept of Gastroenterology,The Second Hospital,Shanxi Medical University,Taiyuan 030001,China;Jiancaoping District Center for Disease Control and Prevention,Taiyuan 030023,China)

机构地区：[1]山西医科大学医学细胞生物学与遗传学教研室,山西太原030001 [2]山西医科大学转化医学研究中心,山西太原030001 [3]山西中医药大学中药与食品工程学院,山西晋中030619 [4]山西医科大学药理学教研室,山西太原030001 [5]山西医科大学第二医院消化内科,山西太原030001 [6]山西省太原市尖草坪区疾病预防控制中心,山西太原030023

出　　处：《中国药理学通报》2023年第5期844-850,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81602176);山西省应用基础研究计划(No 201601D202107);山西省回国留学人员科研资助项目(No 2020-077)。

摘　　要：目的阐明青蒿琥酯在食管鳞癌病理进程中对肿瘤细胞功能以及细胞周期的调控作用。方法用不同浓度青蒿琥酯的培养基处理KYSE450细胞和TE14细胞,以0 mg·L^(-1)青蒿琥酯处理的细胞作为空白组,以30 mg·L^(-1)青蒿琥酯处理后细胞作为实验组。采用MTT和克隆形成实验去测定细胞的生长情况;以Transwell实验去分别检测青蒿琥酯处理前后细胞的侵袭迁移能力的变化情况;利用流式细胞术检测细胞周期变化。通过转录组测序检测基因变化,并利用RT-qPCR验证差异基因以探究青蒿琥酯在ESCC中发挥作用的可能机制。结果与空白组相比,青蒿琥酯处理后KYSE450、TE14细胞的增殖、侵袭、迁移能力明显下降(P<0.01)。测序结果表明,青蒿琥酯可能通过影响细胞周期发挥抑制肿瘤发生发展的作用。青蒿琥酯处理后KYSE450细胞和TE14细胞G 1期比例明显上升。与细胞周期调控密切相关的基因Skp2表达降低,其下游靶基因P21升高。结论青蒿琥酯通过调控Skp2、P21的表达抑制食管鳞癌细胞发生发展。本研究以抗疟疾中成药青蒿琥酯的重新应用为创新点,为开发新型安全、有效的ESCC靶向药物提供了科学依据,也为开发ESCC新治疗方案提供了思路。Aim To clarify the regulatory effect of Artesunate(ART)on tumor cell function and cell cycle in the pathological process of esophageal squamous cell carcinoma(ESCC).Methods KYSE450 and TE14 cells were treated with different concentrations of ART.The cells treated with 0 mg·L^(-1)ART were used as the control group,and the cells treated with 30 mg·L^(-1)ART were used as the experimental group.MTT assay and cloning formation experiment were used to sense cell growth.Transwell assay was applied to test the invasion and migration capacity of ART treated group and blank group separately,flow cytometry was employed to examine cell cycle.The mechanism of Artesunate on ESCC by RNA-seq was detected.Differential genes were validated using RT-qPCR.Results Compared with the control group,the proliferation,invasion and migration of ESCC cells were suppressed considerably after ART treatment(P<0.01).According to the analysis of sequencing results,ART might inhibit the occurrence and development of tumor by affecting the cell cycle.After ART treatment,the ratio of KYSE450 cells at G 1 phase boosted 18.02%.The proportion of KYSE450 cells in G 1 phase increased significantly in the back of adding ART(24.12%±2.62%vs 42.14%±0.65%,P<0.01),and the percentage of TE14 cells at G 1 phase also increased significantly after ART therapy(36.25%±0.21%vs 42.19%±0.38%,P<0.01).Skp2,a gene closely related to cell cycle regulation,decreased and its downstream target gene P21 increased.Conclusions ART,a Chinese patent medicine,can inhibit the proliferation,invasion and migration of ESCC cells,accelerate apoptosis and cause cell arrest through regulating the expression of Skp2 and P21.This study takes the re-application of Chinese patent medicine which is used to against malaria originally as the innovation point to provide a reference basis for the development of new safety,effective and economical targeted drugs for ESCC.It also provides scientific ideas for the development of a new treatment scheme of radiotherapy combined with ESCC.

关 键 词：中成药 青蒿琥酯 食管鳞癌 细胞增殖 细胞迁移 细胞周期 细胞侵袭 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R735.1[医药卫生—基础医学] R735.102.2R916.4