刘晖杰,彭荣,许华
LIU Huijie,PENG Rong,XU Hua

• 1:湖北文理学院附属襄阳市中心医院肿瘤科
• 2:宜城市人民医院肿瘤科
• 3:湖北文理学院附属襄阳市中心医院审计处

摘要(Abstract)：

目的 观察白杨素对肺腺癌顺铂(DDP)耐药性细胞A549/DDP耐药敏感性的影响。方法 取A549和A549/DDP细胞，qRT-PCR及Western blot法检测膜联蛋白A3(Annexin A3)基因及蛋白表达，CCK-8法测不同浓度白杨素(0、5、25、50、100 mmoL/L)干预后细胞生存率；将A549/DDP细胞分为A549/DDP组、白杨素组(50 mol/L)、Annexin A3过表达组(Annexin A3mimic组)、Annexin A3阴性对照组(Annexin A3 NC)、白杨素+Annexin A3mimic组，另取A549细胞正常培养作为空白对照组；CCK-8法检测细胞对DDP的半数生存浓度(IC50)及耐药指数；qRT-PCR和Western blot法检测Annexin A3基因和蛋白表达水平，流式细胞义监测细胞凋亡率，小鼠荷瘤实验检测移植瘤体积。结果 不同浓度的白杨素均能时效依赖性和剂量依赖性降低A549、A549/DDP细胞生存率(P<0.05),选取50 mol/L为实验浓度作用细胞48 h;与A549细胞比较，A549/DDP细胞Annexin A mRNA及蛋白表达、对DDP的IC50、耐药指数均升高，凋亡率降低(P<0.05);与A549/DDP组比较，白杨素组细胞IC50、耐药指数、Annexin A3 mRNA及蛋白表达降低，细胞凋亡率升高(P<0.05),Annexin A3 mimic组细胞IC50、耐药指数、Annexin A3 mRNA及蛋白表达升高，细胞凋亡率降低(P<0.05);Annexin A3基因高表达可减弱A549/DDP细胞对白杨素的敏感性，降低细胞凋亡率(P<0.05);小鼠荷瘤实验证实，与单独的DDP治疗组比较，白杨素与DDP联合治疗可显著降低小鼠A549/DDP移植瘤瘤体体积(P<0.05),Annexin A3基因高表达的小鼠逆转白杨素与DDP联合治疗效果(P<0.05)。结论 白杨素可能通过下调Annexin A3表达，逆转A549/DDP细胞对DDP的耐药性。
Objective To observe the sensitivity of lung adenocarcinoma cisplatin(DDP) resistant cell A549/DDP. Methods A549 and A549/DDP cells were collected and the gene and protein expression of Annexin A3(Annexin A3) were detected by qRT-PCR and Western blot; the cell survival rate was measured by CCK-8 method after different concentrations of chrysin(0, 5, 25, 50, 100 mmoL/L) intervention. The A549/DDP cells were divided into the A549/DDP group, chrysin group(50 mol/L), Annexin A3 overexpression group(Annexin A3mimic group), Annexin A3 negative control group(Annexin A3 NC), chrysin+Annexin A3mimic group, and normal culture of A549 cells as Blank control group. CCK-8 method measured the median survival concentration(IC50) and drug resistance index of cells in the group to DDP. qRT-PCR and Western blot methods were used to detect Annexin A3 gene and protein expression levels; flow cytometry was used to detect cell apoptosis rate; and transplanted tumor volume was detected by mouse tumor-bearing experiment. Results Different concentrations of chrysin reduced the survival rate of A549 and A549/DDP cells in a time-dependent and dose-dependent manner(P<0.05), and 50 mol/L was selected as the experimental concentration to act on the cells for 48 h. Compared with A549 cells, Annexin A mRNA and protein expression, IC50, and drug resistance index of A549/DDP cells were increased and apoptosis rate was reduced(P<0.05). Compared with the A549/DDP group, the IC50, drug resistance index, Annexin A3 gene and protein of the chrysin group were decreased, and the apoptosis rate was increased(P<0.05), and the IC50, drug resistance index, Annexin A3 gene and protein of the Annexin A3 mimic group were increased, and the apoptosis rate was reduced(P<0.05). High expression of Annexin A3 gene attenuates the sensitivity of A549/DDP cells to chrysin and reduces the rate of apoptosis. The mouse tumor-bearing experiment confirmed that compared with the DDP treatment group alone, the combined treatment of chrysin and DDP can significantly reduce the tumor volume of A549/DDP transplanted tumors in mice(P<0.05), and the mice with high Annexin A3 gene expression reversed the effect of the combination of chrysin and DDP treatment(P<0.05). Conclusion Chrysin can reverse the resistance of A549/DDP cells to DDP by down-regulating the expression of Annexin A3.

关键词(KeyWords)： 肺腺肿瘤;白杨素;膜联蛋白A3;顺铂耐药;A549/DDP细胞
lung adenocarcinoma;chrysin;annexin A3;cisplatin resistance;A549/DDP cells

Abstract:

Keywords:

基金项目(Foundation): 湖北省自然科学基金(WJ2019M132)

作者(Author): 刘晖杰,彭荣,许华
LIU Huijie,PENG Rong,XU Hua

DOI: 10.19367/j.cnki.2096-8388.2023.07.009

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