重楼皂苷Ⅱ干预后肺癌细胞增殖、迁移和侵袭能力变化及其与miR-16-5p表达的关系  

Changes in proliferation,migration,and invasion of lung cancer cells after polyphyllinⅡintervention and their relationships with miR-16-5p expression

作　　者：陈琳 刘诗韵 熊琳[1] 冉凤英 刘仁炎 陈琴华 杨光义 朱军[1,2] CHEN Lin;LIU Shiyun;XIONG Lin;RAN Fengying;LIU Renyan;CHEN Qinhua;YANG Guangyi;ZHU Jun(Department of Pharmacy,Sinopharm Dongfeng General Hospital Affiliated to Hubei University of Medicine,Shiyan 442008,China;不详)

机构地区：[1]湖北医药学院附属国药东风总医院药学部,湖北十堰442008 [2]湖北医药学院药学院 [3]深圳市宝安纯中医治疗医院中药临床药学重点实验室

出　　处：《山东医药》2023年第11期1-5,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(82272960);湖北省卫生健康委面上项目(WJ2021M062);湖北省卫生健康委中医药面上项目(ZY2021M038);广东省深圳市科技创新委员基础研究面上项目(JCYJ20210321142012033);湖北省十堰市科技局项目(22Y90)。

摘　　要：目的探讨重楼皂苷Ⅱ对肺癌细胞增殖、迁移和侵袭能力的抑制作用及其可能的机制。方法取对数生长期的肺癌A549细胞,分为高浓度组、中浓度组、低浓度组和对照组,分别加入3、2、1μmol/L重楼皂苷Ⅱ及等量二甲基亚砜溶液,干预24 h。采用CCK-8法检测细胞增殖能力(以细胞增殖活力表示),细胞划痕实验检测细胞迁移能力(以细胞迁移率表示),Transwell侵袭实验检测细胞侵袭能力(以侵袭细胞数表示),Western blotting法检测凋亡相关蛋白[Cleave-Caspase-3、Bax、Bcl-2、基质金属蛋白酶2(MMP-2)]表达,实时荧光定量PCR法检测miR-16-5p表达。取对数生长期的A549细胞,分为miR-16-5p inhibitor组和miR-16-5p NC组和阴性对照组,miR-16-5p inhibitor组和miR-16-5p NC组分别转染含有绿色荧光标记的miR-16-5p inhibitor和miR-16-5p inhibitor NC,并加入2μmol/L重楼皂苷Ⅱ溶液,阴性对照组未进行细胞转染并加入等量二甲基亚砜溶液,干预24 h,参照上法检测miR-16-5p、凋亡相关蛋白表达及细胞生物学行为。结果高、中、低浓度组和对照组细胞增殖活力、细胞迁移率及侵袭细胞数均依次升高,组间两两比较P均<0.05。与对照组比较,高、中、低浓度组Cleave-Caspase-3、Bax、miR-16-5p表达均升高,Bcl-2、MMP-2蛋白表达均降低,以高浓度组变化最明显(P均<0.05)。miR-16-5p inhibitor组、miR-16-5p NC组和阴性对照组miR-16-5p相对表达量分别为0.07±0.05、1.57±0.07、1.00±0.03,miR-16-5p inhibitor组低于miR-16-5p NC组、阴性对照组(P均<0.05)。miR-16-5p NC组细胞增殖活力、细胞迁移率、侵袭细胞数及Bcl-2、MMP-2蛋白表达均低于miR-16-5p inhibitor组和阴性对照组,Cleave-Caspase-3、Bax蛋白表达均高于miR-16-5p inhibitor组和阴性对照组(P均<0.05),miR-16-5p inhibitor组和阴性对照组上述指标比较差异均无统计学意义(P均>0.05)。结论重楼皂苷Ⅱ可抑制肺癌细胞的增殖、迁移和侵袭能力,其机制可能Objective To investigate the inhibitory effects of polyphyllinⅡon the proliferation,migration,and invasion of lung cancer cells and their possible mechanism.Methods Lung cancer A549 cells in the logarithmic growth phase were divided into the high concentration group,medium concentration group,low concentration group,and control group.PolyphyllinⅡof 3,2,1μmol/L and the same amount of dimethyl sulfoxide solution were added for 24 h,respectively.Cell proliferation capacity(represented by cell proliferation activity)was detected by CCK-8.Cell migration ability(expressed as cell mobility)was measured by cell wound healing.Cell invasion ability(represented by the number of cells invaded)was detected by Transwell invasion assay.The expression levels of apoptosis-related proteins[Cleave-Caspase-3,Bax,Bcl-2,matrix metalloproteinase(MMP-2)]were detected by Western blotting.Real-time fluorescence quantitative PCR was used to detect the expression of miR-16-5p.A549 cells in logarithmic growth phase were divided into the miR-16-5p inhibitor group,miR-16-5p NC group,and negative control group.Cells in the miR-16-5p inhibitor group and miR-16-5p NC group were transfected with miR-16-5p inhibitor and miR-16-5p inhibitor NC labeled with green fluores-cence and then were added with 2μmol/L polyphyllinⅡsolution,respectively.Cells in the negative control group did not receive cell transfection and were only added with the same amount of dimethyl sulfoxide solution.At 24 h after interven-tion,the expression of miR-16-5p,apoptosis-related proteins and cell biological behavior were detected by the above meth-ods.Results The cell proliferation viability,cell migration rates and the numbers of invasive cells in the high,medium and low concentration groups and the control group sequentially increased,and significant differences were found between groups(all P<0.05).Compared with the control group,the expression levels of Cleave-Caspase-3,Bax and miR-16-5p in-creased in the high,medium and low concentration groups,and the expressio

关 键 词：重楼皂苷Ⅱ miR-16-5p 细胞增殖 细胞迁移 细胞侵袭 肺癌 

分 类 号：R285.5[医药卫生—中药学]