葡萄糖调节蛋白78在口腔鳞状细胞癌组织中的表达及其对化疗敏感性的影响  

The expression of GRP78 in oral squamous cell carcinoma and its effect on chemotherapeutic sensitivity

作　　者：申铁兵[1] 郭立娜[1] 李琼[1] 夏凤君[1] SHEN Tiebing;GUO Lina;LI Qiong;XIA Fengjun(Department of Stomatology,Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,Inner Mongolia,China)

机构地区：[1]内蒙古医科大学附属医院口腔科,呼和浩特010050

出　　处：《癌症进展》2018年第12期1498-1502,1549,共6页Oncology Progress

摘　　要：目的探讨葡萄糖调节蛋白78(GRP78)在口腔鳞状细胞癌组织中的表达情况及其与口腔鳞状细胞癌细胞株Tca8113对奥沙利铂(L-OHP)敏感性影响的关系,以及可能的作用机制。方法将Tca8113-L-OHP细胞分为未处理组、GRP78 siRNA转染组和阴性对照组。采用免疫组织化学法检测口腔鳞状细胞癌组织中GRP78的相对表达量;采用药物浓度梯度间歇诱导法体外建立Tca8113耐药细胞系;采用噻唑蓝(MTT)法检测L-OHP对Tca8113细胞和Tca8113-L-OHP细胞增殖活性的影响;采用RNA干扰技术靶向沉默Tca8113-L-OHP细胞中GRP78的表达;采用实时荧光定量聚合酶链反应(RT-PCR)和蛋白质印迹法(Western blot)检测Tca8113细胞和Tca8113-L-OHP细胞中GRP78、磷酸化的蛋白酪氨酸激酶2(p-JAK2)、磷酸化的信号转导与转录因子3(p-STAT3)mRNA和p-JAK2、p-STAT3蛋白的相对表达量。结果口腔鳞状细胞癌组织中GRP78的阳性表达率为84.2%,高于正常口腔黏膜组织的20.0%(P﹤0.05);加入L-OHP后,Tca8113细胞中p-JAK2和p-STAT3 mRNA、p-JAK2和p-STAT3蛋白的相对表达量均低于Tca8113-L-OHP细胞(P﹤0.05)。经32μg/ml的L-OHP作用后,Tca8113细胞中p-JAK2、p-STAT3 mRNA和p-JAK2、p-STAT3蛋白的相对表达量均有所下调(P﹤0.05);经不同浓度的L-OHP作用后,GRP78 siRNA组Tca8113-L-OHP细胞的增殖活性明显低于阴性对照组(P﹤0.01)。转染GRP78 siRNA后,Tca8113-L-OHP细胞中p-JAK2、p-STAT3 m RNA和p-JAK2、p-STAT3蛋白的相对表达量均明显低于阴性对照组(P﹤0.01)。结论下调GRP78的表达可降低JAK2和STAT3的磷酸化水平,从而提高口腔鳞状细胞癌细胞对L-OHP的敏感性。Objective To discuss the expressions of glucose regulated protein 78 （GRP78） in oral squamous cell carcinoma （OSCC） tissue,and to explore its effect on the sensitivity to oxaliplatin （L-OHP） of oral squamous cell carcinoma cell line Tca8113 and the possible mechanism. Method The Tca8113-L-OHP cells were divided into untzeated group, GRP78 siRNA transfected group and negative control group. The expressions of GRP78 in OSCC tissues and normal oral tissues were detected by immunohistochemistry. Tca8113-L-OHP cells were built and cultured invitro by drug concentra- tion gradient intermittent induction. The proliferation of Tca8113 cells and Tca8113-L-OHP cells was detected by MTT. The expression of GRP78 was inhibited by small interfering RNA （siRNA）. The relative mRNA expressions of GRP78, phosphorylated protein tyrosine kinase 2 （p-JAK2）, phosphorylated signal transduction and transcription factor 3 （p- STAT3） and the relative protein expression of p-JAK2 and p-STAT3 in Tca8113 cells and Tca8113-L-OHP cells were detected by RT-PCR and western blot.Result The positive expression rate of GRP78 in OSCC tissues （84.2%） was higher than that （20.0%） in the normal tissues （P〈0.05）. After adding L-OHP, the mRNA and protein expressions of p-JAK2, p- STAT3 in Tca8113 cellswere all significantly lower than those in Tca8113-L-OHP cells （P〈0.05）. After treated by 32 μg/ ml L-OHP, the mRNA and protein expressions of p-JAK2, p-STAT3 in Tca8113 cells were significantly decreased （P〈 0.05）. After treated by L-OHP with different concentration, the proliferation activity of Tca8113-L-OHP cells in GRP78 siRNA group were significantly lower than that in negative control group （P〈0.01）. After transfection of GRP78 siRNA, the mRNA and protein expressions of p-JAK2, p-STAT3 in siRNA GRP78 transfected Tca8113-L-OHP cells were signifi- cantly lower than those in negative control cells （P〈0.01）. Conclusion Down-regulation of GRP78 expression could decrease the phosphorylation lev

关 键 词：葡萄糖调节蛋白78 口腔鳞状细胞癌 奥沙利铂 敏感性 JAK/STAT信号通路 

分 类 号：R739.8[医药卫生—肿瘤]