低温等离子体对喉乳头状瘤细胞的影响及机制  

Effect of Non-Thermal Plasma on Laryngeal Papilloma Cells and Its Mechanism

作　　者：王东海 马莹[1] 余文兴 唐梓轩 WANG Donghai;MA Ying;YU Wenxing;TANG Zixuan(Department of Otolaryngology,Suining Central Hospital,Suining Sichuan 629000,China)

机构地区：[1]遂宁市中心医院耳鼻咽喉科,四川遂宁629000

出　　处：《中国医疗设备》2023年第6期146-150,167,共6页China Medical Devices

基　　金：遂宁市中心医院科研项目(20210526)。

摘　　要：目的探讨低温等离子体(Non-Thermal Plasma,NTP)对人喉乳头状瘤细胞的影响及机制。方法人喉乳头状瘤细胞株Hs840.T重悬细胞后分为A组(正常培养的Hs840.T细胞株)、B组(正常培养的Hs840.T细胞株经NTP干预2 min)及C组(正常培养的Hs840.T细胞株经NTP干预4 min)。采用细胞计数试剂盒-8方法检测各组细胞存活率;流式细胞仪检测各组细胞凋亡率;免疫印迹检测各组细胞中表皮生长因子受体(Epidermal Growth Factor Receptor,EGFR)、单克隆血管内皮生长因子(Vascular Endothelial Growth Factor,VEGF)、B细胞淋巴瘤2(B-Cell Lymphoma 2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2-Associated X Protein,Bax)及半胱氨酸天冬氨酸蛋白酶3(Cysteine-Aspartic Acid Protease 3,Caspase-3)蛋白水平;定量反转录聚合酶链式反应检测各组细胞miRNA-196a和miRNA-196b表达。结果与A组相比,B组和C组24、48、72、96 h时细胞活性均显著降低(P<0.05)。A组、B组及C组Hs840.T细胞凋亡率比较存在显著差异(P<0.05)。与A组相比,B组和C组Hs840.T细胞中Bcl-2表达升高,EGFR、VEGF、Bax及Caspase-3、miRNA-196a及miRNA-196b水平显著降低(P<0.05);与B组相比,C组Hs840.T细胞中Bcl-2表达显著升高,EGFR、VEGF、Bax、Caspase-3、miRNA-196a及miRNA-196b水平显著降低(P<0.05)。结论NTP对人喉乳头状瘤细胞杀伤较强,伴随时间依赖性,可以调节EGFR、VEGF、Bcl-2、Bax及Caspase-3蛋白活性而加快肿瘤细胞凋亡,研究机制可能与抑制miRNA-196a表达相关。Objective To investigate the effect of non-thermal plasma(NTP)on human laryngeal papilloma cells and its mechanism.Methods The human laryngeal papilloma cell line Hs840.T was resuspended and divided into three groups:group A(normal Hs840.T cells under routine culture conditions),group B(Hs840.T cells intervened with NTP for 2 minutes under normal culture conditions),and group C(Hs840.T cells intervened with NTP for 4 minutes under normal culture conditions).Cell viability rate of each group was detected by cell count Kit 8 method;flow cytometry was used to detect cell apoptosis rate in each group.Epidermal growth factor receptor(EGFR),monoclonal vascular endothelial growth factor(VEGF),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax)and cysteine-aspartic acid protease 3(Caspase-3)protein level were detected by western blotting.Quantitative reverse transcription polymerase chain reaction detected the expressions of miRNA-196a and miRNA-196b in each group.Results Compared with group A,the cell viability of groups B and C was significantly decreased at 24 h,48 h,72 h and 96 h(P<0.05).The apoptosis rate of Hs840.T cells in groups A,B and C were significantly different(P<0.05).Compared with group A,Bcl-2 in Hs840.T cells of group B and group C was increased,and the levels of EGFR,VEGF,Bax,Caspase-3,miRNA-196a and miRNA-196b were significantly decreased(P<0.05).Compared with group B,Bcl-2 in Hs840.T cells of group C was significantly increased,and the levels of EGFR,VEGF,Bax,Caspase-3,miRNA-196a and miRNA-196b were significantly decreased(P<0.05).Conclusion NTP has a strong killing effect on human laryngeal papilloma cells with a time-dependent effect,which can regulate the activities of EGFR,VEGF,Bcl-2,Bax and Caspase-3 proteins so as to accelerate the apoptosis of tumor cells,the mechanism may be related to the inhibition of miRNA-196a expression.

关 键 词：人喉乳头状瘤 低温等离子体 miRNA-196a 上皮生长因子受体 血管内皮生长因子 

分 类 号：R762[医药卫生—耳鼻咽喉科]