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李小娜,李开瑞,何迎春,徐朝军,宋岚.血根碱抑制肺腺癌A549细胞生长及机制初探[J].湖南中医药大学学报,2021,41(1):61-66[点击复制] |
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| 血根碱抑制肺腺癌A549细胞生长及机制初探 |
| 李小娜,李开瑞,何迎春,徐朝军,宋岚 |
| (湖南中医药大学医学院, 湖南 长沙 410208;湖南中医药大学医学院, 湖南 长沙 410208;中医药防治眼耳鼻喉疾病湖南省重点实验室, 湖南 长沙 410208;湖南省中医药防治眼耳鼻喉疾病与视功能保护工程技术研究中心, 湖南 长沙 410208;湖南中医药大学第一附属医院心胸外科, 湖南 长沙 410007;湖南中医药大学中医学国内一流建设学科, 湖南 长沙 410208) |
| 摘要: |
| 目的 研究血根碱（sanguinarine，SAN）对肺腺癌A549细胞增殖、凋亡和细胞周期的影响，并探讨其可能的作用机制。方法 将肺腺癌A549细胞随机分为空白组、SAN不同浓度（1.25、2.5、5、10 μmol/L）组、阳性组。采用噻唑蓝[3-（4，5-dimethyl-2-thiazolyl）-2，5-diphenyl-2-H-tetrazolium bromide，thiazolyl blue tetrazolium bromide，MTT]法和实时无标记动态细胞分析技术（real time cellular analysis，RTCA）检测细胞增殖；采用流式细胞术检测细胞周期分布；采用Annexin V-FITC/PI双荧光染色法检测细胞凋亡；采用蛋白免疫印迹（Western blot）法检测增殖相关蛋白（PCNA）、周期相关蛋白（Cyclin D1、Cyclin D3、CDK4）、凋亡相关蛋白（Bax、XIAP、Survivin）表达水平。结果 MTT法和RTCA检测结果均显示，不同浓度SAN均能够抑制肺腺癌A549细胞增殖（P<0.01）。流式细胞术检测结果显示，不同浓度SAN（2.5、5 μmol/L）处理肺腺癌A549细胞24 h后，其G1期比例显著增加（P<0.01）。AnnexinV-FITC/PI双荧光染色法结果，经SAN（2.5、5 μmol/L）处理24 h后的肺腺癌A549细胞凋亡率明显增加（P<0.01）。Western blot结果显示，经SAN干预后的肺腺癌A549细胞的增殖相关蛋白PCNA（P<0.01）、周期相关蛋白Cyclin D1、Cyclin D3、CDK4（P<0.01）、抗凋亡蛋白Survivin、XIAP（P<0.05或P<0.01）表达水平均显著降低，促凋亡蛋白Bax（P<0.01）表达水平明显提高。结论 SAN能抑制肺腺癌A549细胞增殖，将其细胞周期阻滞于G1期，并可诱导其细胞凋亡。 |
| 关键词: 血根碱 肺腺癌A549细胞 增殖 凋亡 细胞周期 |
| DOI：10.3969/j.issn.1674-070X.2021.01.011 |
| 投稿时间：2020-09-25 |
| 基金项目:国家自然科学基金项目（81874408）；中国博士后科学基金项目（2015M580690）；湖南省自然科学基金项目（2019JJ40216）；湖南省中医药科研计划项目（201696）；中医内科学省部共建教育部重点实验室开放基金项目（ZYNK201506）；湖南省干细胞中药调控实验室开放基金项目（2013GXB02）；湖南省研究生创新项目（CX2018B482）；湖南中医药大学研究生创新项目（2018CX62）；湖南省研究生科研创新项目（CX20190552）。 |
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| Inhibitory Effect of Sanguinarine on the Growth of Lung Adenocarcinoma A549 Cells and its Mechanism |
| LI Xiaona,LI Kairui,HE Yingchun,XU Zhaojun,SONG Lan |
| (School of Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;School of Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Provincial Key Laboratory for the Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Traditional Chinese Medicine, Changsha, Hunan 410208, China;Hunan Provincial Ophthalmology and Otolaryngology Diseases Prevention and Treatment with Traditional Chinese Medicine and Visual Function Protection Engineering and Technological Research Center, Changsha, Hunan 410208, China;Department of Cardiothoracic Surgery, The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;The Domestic First-class Discipline Construction Project of Chinese Medicine of Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To investigate the effects of sanguinarine (SAN) on proliferation, apoptosis and cell cycle of lung adenocarcinoma A549 cells and to explore its mechanism. Methods Lung adenocarcinoma A549 cells were randomly divided into the blank group, different concentrations of SAN (1.25, 2.5, 5, 10 μmol/L) groups and the positive group. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,thiazolyl blue tetrazolium bromide (MTT) and real-time cellular label free dynamic analysis (RTCA) were used to detect the cell proliferation. Flow cytometry was performed to evaluate the cell cycle distribution. Annexin V-FITC/PI double fluorescent staining was used to detect the cell apoptosis. Western blot was used to detect the expression levels of proliferation-related protein (PCNA), cell cycle-related proteins (Cyclin D1, Cyclin D3, CDK4), and apoptosis-related proteins (Bax, XIAP, Survivin) Results The results of MTT and RTCA showed that SAN at different concentrations could inhibit the proliferation of lung adenocarcinoma A549 cells (P<0.01). Flow cytometry showed that the G1 phase proportion of lung adenocarcinoma A549 cells was significantly increased (P<0.01) after treated with different concentrations of SAN (2.5, 5 μmol/L) for 24 h. Annexin V-FITC/PI double fluorescent staining showed that the apoptosis rate was significantly increased (P<0.01) after treated with SAN (2.5, 5 μmol/L) for 24 h. Western blot showed that, the expression of proliferation-related protein PCNA (P<0.01), cell cycle-related protein Cyclin D1, Cyclin D3 and CDK4 (P<0.01), and anti apoptotic protein Survivin and XIAP (P<0.05 or P<0.01) were decreased, while the expression of pro-apoptotic protein Bax was significantly increased (P<0.01). Conclusion SAN can inhibit the proliferation and arrest the cell cycle in G1 phase, and induce apoptosis of lung adenocarcinoma A549 cells. |
| Key words: sanguinarine lung adenocarcinoma A549 cells proliferation apoptosis cell cycle |
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