舒尼替尼诱导自然杀伤细胞2族成员D配体表达对自然杀伤细胞杀伤舌鳞癌CAL27细胞敏感性的影响  

Effect of sunitinib-induced natural killer group 2 member D ligand expression on killing sensitivity of natural killer cells to tongue squamous cell carcinoma CAL27 cells

作　　者：朱冠宇 李阿峰[2] 朱含九 李楠 ZHU Guan-yu;LI A-feng;ZHU Han-jiu;LI Nan(Department of Stomatology,Qionghai People′s Hospital,Qionghai 571400,China;Department of Stomatology,Shangluo Central Hospital,Shangluo 726000,China;Department of Stomatology,Shangnan County Hospital of Traditional Chinese Medicine,Shangnan 726300,China;School of Basic Medicine and Life Sciences,Hainan Medical College,Haikou 571199,China)

机构地区：[1]海南省琼海市人民医院口腔科,琼海市571400 [2]陕西省商洛市中心医院口腔科,商洛市726000 [3]陕西省商南县中医医院口腔科,商南县726300 [4]海南医学院基础医学与生命科学学院,海口市571199

出　　处：《广西医学》2021年第20期2441-2446,共6页Guangxi Medical Journal

基　　金：海南省卫生健康行业科研项目(19A200039)。

摘　　要：目的探讨舒尼替尼诱导自然杀伤细胞2族成员D(NKG2D)配体(NKG2DL)的表达对自然杀伤(NK)细胞杀伤舌鳞癌CAL27细胞敏感性的影响。方法将对数生长期的CAL27细胞分为未处理组和舒尼替尼组,舒尼替尼组采用舒尼替尼处理,未处理组细胞未经舒尼替尼处理。采用细胞克隆形成实验检测两组CAL27细胞的增殖能力;采用流式细胞术检测两组CAL27细胞的凋亡、NKG2DL(MICA、MICB、ULBP1、ULBP2和ULBP3)阳性细胞的表达及健康人NK细胞的纯度;采用免疫蛋白印迹法检测两组CAL27细胞中NKG2DL(MICA、MICB、ULBP1、ULBP2和ULBP3)蛋白的表达水平。再将舒尼替尼组分为舒尼替尼-NKG2D组和舒尼替尼-对照组,舒尼替尼-NKG2D组加入经NKG2D单抗处理后的NK细胞,舒尼替尼-对照组加入未经NKG2D单抗处理的NK细胞,同时未处理组加入未经NKG2D单抗处理的NK细胞(作为未处理-NK组)。采用乳酸脱氢酶释放法检测NK细胞对CAL27细胞的杀伤敏感性。结果(1)与未处理组相比,舒尼替尼组CAL27细胞的增殖能力降低、凋亡率增加,CAL27细胞中的MICA、MICB、ULBP1和ULBP2蛋白表达水平均升高(均P<0.05),而两组CAL27细胞ULBP3蛋白表达水平差异无统计学意义(P>0.05)。(2)与未处理-NK组相比,NK细胞对经舒尼替尼处理的CAL27细胞的杀伤敏感性增强(均P<0.05)。(3)采用NKG2D抗体封闭NK细胞表面的NKG2D受体后,NK细胞对经舒尼替尼处理的CAL27细胞的杀伤敏感性低于未采用NKG2D单抗处理的CAL27细胞的杀伤敏感性(均P<0.05)。结论舒尼替尼可以抑制舌鳞癌细胞CAL27细胞的增殖,促进其凋亡,上调CAL27细胞中NKG2DL(MICA、MICB、ULBP1、ULBP2)的表达水平,增强NK细胞对CAL27细胞的杀伤敏感性。Objective To investigate the effect of sunitinib-induced natural killer group 2 member D ligand(NKG2DL)expression on the killing sensitivity of natural killer(NK)cells to tongue squamous cell carcinoma CAL27 cells.Methods CAL27 cells in logarithmic phase were divided into non-treatment group and sunitinib group.The sunitinib group received sunitinib for treatment,while the non-treatment group did not receive sunitinib for treatment.In both groups,cell colony formation assay was used to detect the proliferation ability of CAL27 cells;flow cytometry was employed to detect the apoptosis of CAL27 cells,the expression of NKG2DL(MICA,MICB,ULBP1,ULBP2 and ULBP3)-positive cells,and the purity of healthy people′s NK cells;Western blotting assay was used to detect the expression level of NKG2DL(MICA,MICB,ULBP1,ULBP2 and ULBP3)protein in CAL27 cells.Then the sunitinib group was further divided into sunitinib-NKG2D group and sunitinib-control group.The NK cells treated with NKG2D monoclonal antibody were added into the sunitinib-NKG2D group,while the NK cells not treated with NKG2D monoclonal antibody were added into the sunitinib-control group.At the same time,the NK cells not treated with NKG2D monoclonal antibody were added into the non-treatment group(non-treatment-NK group).Lactate dehydrogenase releasing assay was adopted to determine the killing sensitivity of NK cells to CAL27 cells.Results(1)Compared with the non-treatment group,the sunitinib group exhibited a decreased proliferation ability and an increased apoptosis rate of CAL27 cells,together with elevated expression levels of MICA,MICB,ULBP1,and ULBP2 proteins in CAL27 cells(all P<0.05),whereas there was no statistically significant difference in the expression level of ULBP3 protein in CAL27 cells between the two groups(P>0.05).(2)Compared with the non-treatment-NK group,NK cells had an increased killing sensitivity to sunitinib-treated CAL27 cells(all P<0.05).(3)NK cells harbored a lower killing sensitivity to sunitinib-treated CAL27 cells than to CAL27 cel

关 键 词：舒尼替尼 自然杀伤细胞2族成员D 自然杀伤细胞 舌鳞癌 

分 类 号：R739.86[医药卫生—肿瘤]