大蒜素对子宫内膜癌Ishikawa细胞顺铂耐药株增殖和凋亡及侵袭与迁移的影响  

Effects of allicin on the proliferation,apoptosis,invasion and metastasis of cisplatin-resistant strains of endometrial carcinoma Ishikawa cells

作　　者：汪晶 李新[1] 周筠[1] 黄金玲[1] 洪莉[1] WANG Jing;LI Xin;ZHOU Yun;HUANG Jinling;HONG Li(Department of Gynecology,People's Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院妇科,湖北武汉430060

出　　处：《中华肿瘤防治杂志》2023年第9期513-519,共7页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金面上项目(61772376)。

摘　　要：目的探讨大蒜素对子宫内膜癌顺铂(DDP)耐药细胞株Ishikawa/DDP生物学行为的影响。方法建立子宫内膜癌DDP耐药细胞株,设对照组、DDP处理组、低剂量大蒜素+DDP处理组、中剂量大蒜素+DDP处理组和高剂量大蒜素+DDP处理组,通过细胞计数实验(CCK8)、Transwell实验、划痕实验、流式细胞术检测各组细胞的增殖、侵袭、迁移和凋亡,采用RT-qPCR实验、蛋白质印迹实验分别检测各组细胞中多药耐药基因1(MDR1)mRNA、P-gp蛋白表达水平。结果低剂量大蒜素+DDP处理组、中剂量大蒜素+DDP处理组和高剂量大蒜素+DDP处理组细胞增殖抑制率分别为(20.92±3.28)%、(31.71±2.82)%和(47.34±3.43)%,随着大蒜素浓度增加,细胞增殖抑制率上升,差异有统计学意义,F=346.231,P=0.007。Transwell实验表明,对照组、DDP处理组、低剂量大蒜素+DDP处理组、中剂量大蒜素+DDP处理组和高剂量大蒜素+DDP处理组侵袭细胞数分别为(314.00±7.53)、(278.00±6.52)、(203.00±5.46)、(109.00±5.32)和(65.00±3.41)个,随着大蒜素浓度增加,细胞侵袭力下降,差异有统计学意义,F=437.807,P=0.009。对照组、DDP处理组、低剂量大蒜素+DDP处理组、中剂量大蒜素+DDP处理组和高剂量大蒜素+DDP处理组48 h划痕宽度相对比例分别为(31.48±9.28)%、(33.76±7.43)%、(42.76±5.34)%、(53.56±7.45)%和(64.431±8.32)%,随着大蒜素浓度增加,细胞迁移能力下降,差异有统计学意义,F=345.126,P=0.005。流式细胞术结果表明,对照组、DDP处理组、低剂量大蒜素+DDP处理组、中剂量大蒜素+DDP处理组和高剂量大蒜素+DDP处理组细胞凋亡率分别为(11.32±1.13)%、(13.51±0.83)%、(21.30±1.56)%、(37.64±2.21)%和(51.31±2.42)%,随着大蒜素浓度增加,细胞凋亡率显著提高,组间均值差异有统计学意义,F=423.012,P=0.008。RT-qPCR结果显示,对照组、DDP处理组、低剂量大蒜素+DDP处理组、中剂量大蒜素+DDP处理组和高剂量大蒜素+DDP处理组MDRObjective To investigate the effect of allicin on the biological behavior of endometrial cancer cisplatin(DDP)-resistant cell line Ishikawa/DDP.Methods Cisplatin-resistant cell strains cultures of endometrial carcinoma were respectively constituted into control,cisplatin-treated,(low-dose allicin+cisplatin)-treated,(medium-dose allicin+cisplatin)-treated,and(high-dose allicin+cisplatin)-treated groups.The proliferation,invasion,metastasis and apoptosis of the cells in each group were detected by CCK8,transwell assay,scratch assay and flow cytometry,and the expression levels of MDR1 mRNA and P-gp protein in each group were detected by RT-qPCR and western blot assay,respectively.Results The inhibition rates of cell proliferation in the low-dose allicin+DDP-treated group,the medium-dose allicin+DDP-treated group and the high-dose allicin+DDP-treated group were(20.92±3.28)%,(31.71±2.82)%and(47.34±3.43)%,respectively,and the inhibition rates of cell proliferation increased with the increase of allicin concentration,and the difference was significant(F=346.231,P=0.007).Transwell experiments showed that the number of invading cells in the control,DDP-treated,low-dose allicin+DDP-treated,medium-dose allicin+DDP-treated and high-dose allicin+DDP-treatedgroupswere(314.00±7.53),(278.00±6.52),(203.00±5.46),(109.00±5.32)and(65.00±5.32),respectively.The difference was statistically significant(F=437.807,P=0.009).The scratch test showed that the relative proportions of scratch widths after 48 h in the control,DDP-treated,low-dose allicin+DDP-treated,medium-dose allicin+DDP-treated and high-dose allicin+DDP-treated groups were(31.48±9.28)%,(33.76±7.43)%,(42.76±5.34)%,(53.56±7.45)%and(64.43±8.32)%,respectively,and the cell migration capacity decreased with increasing allicin concentration,with statistically significant differences(F=345.126,P=0.005).The flow cytometry results showed that the apoptosis rates in the control,DDP-treated,low-dose allicin+DDP-treated,medium-dose allicin+DDPtreated and high-dose allicin+DD

关 键 词：子宫内膜癌 大蒜素 DDP耐药 多药耐药基因1 P-糖蛋白 

分 类 号：R737.33[医药卫生—肿瘤]