热疗联合吉西他滨对舌鳞癌细胞生物学行为影响的研究  

Study on the effects of thermotherapy combined with gemcitabine on the biological behavior of tongue squamous cell carcinoma cells

作　　者：邵云 丛瑗 李受益 王玮[1,2] 杨玉莹 周学筱 王昇志 郝玉丽 Shao Yun;Cong Yuan;Li Shouyi;Wang Wei;Yang Yuying;Zhou Xuexiao;Wang Shengzhi;Hao Yuli(School of Stomatology,Qingdao University,Qingdao 266003,China;Department of Stomatology,The Afiliated Yantai Yuhuangding Hospital of Qingdao University,Yantai 264000,China;Department of Dental Implantology,West China Hospital of Stomatology Sichuan University,Chengdu 610044,China)

机构地区：[1]青岛大学口腔医学院,青岛266023 [2]青岛大学附属烟台毓璜顶医院口腔科,烟台264000 [3]四川大学华西口腔医院口腔种植科,成都610044

出　　处：《中华放射肿瘤学杂志》2024年第9期853-858,共6页Chinese Journal of Radiation Oncology

摘　　要：目的探讨热疗联合吉西他滨对舌鳞癌细胞生物学行为的影响。方法将人舌鳞癌细胞Tca8113分为对照组(空白对照)、吉西他滨组、热疗组(43℃培养箱中加热1 h后再在37℃培养箱中孵育24 h)、热疗+吉西他滨组。EdU染色、CCK-8检测Tca8113细胞增殖能力;流式细胞术检测Tca8113细胞凋亡和细胞周期;Transwell检测Tca8113细胞侵袭;Western blot检测Tca8113细胞中细胞周期蛋白D1(CyclinD1)、Bcl-2相关X蛋白(Bax)、基质金属蛋白酶(MMP)-9、磷酸化组蛋白H2AX(γH2AX)、Nijmegen断裂综合征1(NBS1)蛋白表达。体内裸鼠移植瘤试验检测移植瘤质量与体积变化。单因素方差分析评估组间差异, 两两比较用SNK-q检验。结果与对照组相比, 吉西他滨组、热疗组、热疗+吉西他滨组Tca8113细胞EdU阳性细胞百分比、OD450值、侵袭细胞数、CyclinD1、MMP-9、NBS1蛋白表达降低, 细胞凋亡率、Bax、γH2AX蛋白表达升高(q=4.45~72.06, P<0.001);与对照组相比, 吉西他滨组、热疗+吉西他滨组G0/G1期细胞比例降低, S、G2/M期细胞比例升高, 热疗组G0/G1期细胞比例降低, G2/M期细胞比例升高(q=10.36~61.09, P<0.001);与吉西他滨组、热疗组相比, 热疗+吉西他滨组Tca8113细胞EdU阳性细胞百分比、OD450值、G0/G1期细胞比例、侵袭细胞数, 以及CyclinD1、MMP-9、NBS1蛋白表达降低, 细胞凋亡率、S期和G2/M期细胞比例、Bax和γH2AX蛋白表达升高(q=4.45~28.73, P<0.001)。体内裸鼠移植瘤试验显示, 与裸鼠对照组相比, 裸鼠吉西他滨组、裸鼠热疗组、裸鼠热疗+吉西他滨组移植瘤体积和质量降低(q=5.58~73.02, P<0.001);与裸鼠吉西他滨组、裸鼠热疗组相比, 裸鼠热疗+吉西他滨组移植瘤体积和质量降低(q=5.58~21.45, P<0.001)。结论热疗与吉西他滨二者联合可抑制Tca8113细胞增殖、侵袭, 阻滞细胞周期, 并诱导细胞凋亡。Objective To explore the impacts of thermotherapy combined with gemcitabine on the biological behavior of tongue squamous cell carcinoma cells.Methods Human tongue squamous cell carcinoma Tca8113 cells were divided into the control group(blank control),gemcitabine group,thermotherapy group(heated in an incubator at 43℃for 1 h and then incubated at 37℃for 24 h)and thermotherapy+gemcitabine group.The proliferation ability of Tca8113 cells was assessed by EdU staining and CCK-8 assay.Cell apoptosis and cell cycle of Tca8113 cells were detected by flow cytometry.The invasion of Tca8113 cells was determined by Transwell chamber assay.The expression levels of cyclin D1(CyclinD1),Bcl-2-associated X protein(Bax),matrix metalloproteinase(MMP)-9,phosphorylated histone H2AX(γH2AX)and Nijmegen breakage syndrome 1(NBS1)proteins in Tca8113 cells were measured by Western blot.The changes of tumor mass and volume were detected by xenograft tumor in vivo test in nude mice.Multi-group comparison was performed by one-way ANOVA.Two group comparison was conducted by SNK-q test.Results Compared with the control group,EdU positive cell percentage,OD450 value,invasive cell number,CyclinD1,MMP-9 and NBS1 protein expression of Tca8113 cells were decreased,whereas the apoptosis rate,the expression of Bax andγH2AX proteins were increased in the gemcitabine,thermotherapy and thermotherapy+gemcitabine groups(q=4.45-72.06,all P<0.001).Compared with the control group,the proportion of G0/G1 phase cells was decreased,whereas the proportion of S and G2/M phase cells was increased in the gemcitabine and thermotherapy+gemcitabine groups,the proportion of G0/G1 phase cells was decreased and the proportion of G2/M phase cells was increased in the hyperthermia group(q=10.36-61.09,all P<0.001).Compared with the gemcitabine and thermotherapy groups,EdU positive cell percentage,OD450 value,G0/G1 phase cell proportion,invasive cell number,CyclinD1,MMP-9 and NBS1 protein expression of Tca8113 cells were decreased,whereas apoptosis rate,S,G2/M phase c

关 键 词：高温 诱发 热疗法 吉西他滨 舌鳞状细胞癌 增殖 细胞周期 凋亡 

分 类 号：R739.86[医药卫生—肿瘤]