谷胱甘肽S-转移酶P1对小鼠肺癌细胞放射敏感性调控作用  

Regulation of glutathione S-transferase P1 on the radiosensitivity of mouse Lewis lung cancer cells

作　　者：梁延杰 张沛 杜乐辉[1] 马娜[1] 雷霄 韩亚楠 赵鑫尧 曲宝林[1] Liang Yanjie;Zhang Pei;Du Lehui;Ma Na;Lei Xiao;Han Yanan;Zhao Xinyao;Qu Baolin(Department of Radiation Oncology,First Medical Center of PLA General Hospital,Beijing 100853,China;PLA Medical School,Beijing 100853,China)

机构地区：[1]解放军总医院第一医学中心放疗科,北京100853 [2]解放军医学院,北京100853

出　　处：《中华放射肿瘤学杂志》2021年第5期498-502,共5页Chinese Journal of Radiation Oncology

基　　金：军委后勤保障部卫生局保健课题(20BJZ40);国家老年疾病临床医学研究中心开放项目(ZXD2004)。

摘　　要：目的探讨谷胱甘肽S-转移酶P1(GSTP1)对小鼠路易斯肺癌细胞放射敏感性的调控作用。方法使用shRNA干扰表达慢病毒和阴性对照病毒分别感染路易斯肺癌(LLC)细胞,并筛选出稳转株;应用RT-PCR和蛋白免疫印迹(WB)法检测LLC细胞GSTP1 mRNA和蛋白质表达水平,验证敲低效果;使用细胞增殖检测(CCK-8)法检测照射后细胞活力;使用克隆形成实验检测照射后细胞克隆形成能力;使用流式细胞术检测照射后细胞的凋亡水平。构建荷瘤小鼠并进行照射,检测照射后肿瘤体积变化;TUNEL染色检测照射后肿瘤细胞凋亡水平;免疫荧光检测照射后肿瘤内CD4+CD8+T细胞数量。结果RT-PCR和WB结果显示shRNA慢病毒干扰后,GSTP1的mRNA和蛋白表达水平均明显下降。下调GSTP1能降低照射后细胞活力及克隆形成能力,并提高照射后细胞凋亡率。GSTP1敲低组荷瘤小鼠照射后肿瘤体积明显小于对照组,而肿瘤凋亡水平高于对照组,肿瘤内浸润CD4+CD8+T细胞数量亦高于对照组。结论下调GSTP1能够显著增加LLC细胞的放射敏感性,并增加肿瘤组织中淋巴细胞的浸润。Objective To explore the regulatory effect of glutathione S-transferase P1(GSTP1)on the radiosensitivity of mouse Lewis lung cancer(LLC)cells.Methods GSTP1-shRNA lentivirus and negative control lentivirus were used to respectively infect the LLC cells,and stable transgenic strains were selected.Real-time PCR and Western blot were conducted to quantitatively measure the expression levels of GSTP1 mRNA and protein in the LLC cells to verify the knockdown effect.The cell counting kit-8(CCK-8)assay was used to detect cell viability after irradiation.The colony formation assay was utilized to assess the cell proliferation ability after irradiation.Flow cytometry was performed to assess the level of cell apoptosis after irradiation.The tumor-bearing mice were established and irradiated to detect the changes in the tumor volume after irradiation.TUNEL staining was employed to detect the level of tumor apoptosis after irradiation.Immunofluorescence was used to detect the number of CD4+CD8+T cells in the tumor after irradiation.Results Real-time PCR and Western blot showed that after shRNA lentivirus interference,the expression levels of GSTP1 mRNA and protein were significantly down-regulated.Down-regulation of GSTP1 reduced cell viability and proliferation,and increased the rate of cell apoptosis after irradiation.The tumor volume of the tumor-bearing mice after irradiation in the GSTP1 knockdown group was significantly smaller than that in the NC group,whereas the tumor apoptosis rate was significantly higher and the number of infiltrating CD4+CD8+T cells in the tumor was remarkably higher compared with those in the control group.Conclusion Knockdown of GSTP1 can significantly increase the radiosensitivity of LLC cells and enhance the infiltration of lymphocytes in tumor tissues.

关 键 词：谷胱甘肽S-转移酶P1 肺癌细胞系 放射敏感性 

分 类 号：R734.2[医药卫生—肿瘤]