山奈素通过下调circ-RPL15抑制肺癌细胞A549的增殖和迁移  

Kaempferide inhibits the proliferation and migration of lung cancer cell A549 by down-regulating circ-RPL15

作　　者：郑晟超 毛佳斌[1] 何大强 ZHENG Chengchao;MAO Jiabin;HE Daqiang(Department of Tuberculosis,the Affiliated Hospital of Hangzhou Normal University,Hangzhou 310015,China;Department of Laboratory,Hangzhou First People’s Hospital Affiliated to Medical College of Zhejiang University,Hangzhou 310006,China)

机构地区：[1]杭州师范大学附属医院结核科,浙江杭州310015 [2]浙江大学医学院附属杭州市第一人民医院检验科,浙江杭州310006

出　　处：《温州医科大学学报》2021年第12期960-967,共8页Journal of Wenzhou Medical University

基　　金：浙江省医药卫生科技计划项目(2019ZD014)。

摘　　要：目的:探索山奈素是否通过circ-RPL15影响肺癌细胞A549的生物行为。方法:使用25、50和75μg/m L山奈素处理肺癌细胞A549。在细胞A549中沉默circ-RPL15,或过表达circ-RPL15并使用75μg/m L山奈素处理。采用细胞计数试剂盒8(CCK-8)检测各组肺癌细胞A549活力变化,平板克隆法检测细胞克隆形成,流式细胞术检测细胞凋亡,蛋白质印迹(Western blot)法检测裂解-含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved-caspase3)蛋白的表达,使用Transwell法检测细胞迁移,并使用实时定量聚合酶链反应(RT-q PCR)检测circ-RPL15、mi R-489-3p表达。双荧光素酶报告实验分析circ-RPL15对miR-489-3p的靶向调控。结果:与对照组相比,不同质量浓度(25、50和75μg/mL)山奈素作用于肺癌细胞A549后,细胞活力、克隆形成数、迁移数、circ-RPL15表达量降低,而凋亡率、Cleaved-caspase3蛋白和miR-489-3p的表达量升高(P <0.05),且均呈浓度依赖性。沉默circ-RPL15增强肺癌细胞A549的miR-489-3p表达量、凋亡率和Cleaved-caspase3蛋白的表达量,并减弱细胞活力、克隆形成数、迁移数(P<0.05)。circ-RPL15靶向调控mi R-489-3p。过表达circ-RPL15逆转了山奈素作用的肺癌细胞增殖、凋亡和迁移情况(P<0.05)。结论:山奈素通过下调circ-RPL15并靶向调控miR-489-3p,抑制肺癌细胞的增殖和迁移,并诱导其凋亡。Objective: To explore whether kaempferide can affect the biological behavior of lung cancer cell A549 through circ-RPL15. Methods: Kaempferide of 25, 50, and 75 μg/mL was used to treat lung cancer cells A549. The dual luciferase report experiment was used to analyze the targeted regulation of miR-489-3 p by circ-RPL15. circ-RPL15 was silenced or overexpressed in cell A549, and treated with 75 μg/mL kaempferol. Cell counting kit-8(CCK-8) was applied to measure changes in the viability of lung cancer cells A549 in each group;plate cloning was performed to detect cell clone formation;flow cytometry was used to monitor cell apoptosis;Western blot was employed to detect the expression of Cleaved-caspase3 protein;Transwell method was used to detect cell migration, and real-time quantitative polymerase chain reaction(RT-qPCR) to determine the expression of circ-RPL15 and miR-489-3 p. The dual luciferase report experiment analyzed the targeted regulation of miR-489-3 p by circ-RPL15. Results: Compared with the control group, after different concentrations(25, 50 and 75 μg/mL) of kaempferide acted on lung cancer cell A549, cell viability, clone formation number, migration number and circ-RPL15 expression decreased, while apoptosis rate, Cleaved-caspase3 protein and miR-489-3 p expression increased(P<0.05), and all were concentration-dependent. Silencing circ-RPL15 enhanced the expression of miR-489-3 p, apoptosis rate and Cleaved-caspase3 protein expression of lung cancer cell A549, and weakened cell viability, the number of clone formation, and the number of migration(P<0.05). circ-RPL15 targets miR-489-3 p. Overexpression of circ-RPL15 reversed the proliferation, apoptosis and migration of lung cancer cells that were affected by kaempferide(P<0.05). Conclusion: Kaempferide inhibits the proliferation and migration of lung cancer cells and induces their apoptosis by down-regulating circ-RPL15 and targeting miR-489-3 p.

关 键 词：肺癌细胞A549 山奈素 细胞迁移 细胞凋亡 circ-RPL15 mi R-489-3p 

分 类 号：R734.2[医药卫生—肿瘤]