黏着斑激酶在口腔鳞癌组织中的表达及其与肿瘤细胞恶性细胞行为的关系

Expression of focal adhesion kinase in oral squamous cell carcinoma and its relationship with malignant behavior of tumor cells

目的:探讨黏着斑激酶(FAK)在口腔鳞癌组织中的表达及其与肿瘤细胞迁移和侵袭的关系。方法:选取新乡医学院第一附属医院和新乡医学院第三附属医院2015年9月至2018年9月收集的79例口腔鳞癌组织和癌旁组织作为研究标本,采用免疫组织化学分析癌旁组织和口腔鳞癌组织FAK蛋白的表达水平;采用FAK短发卡RNA(shRNA)和对照shRNA慢病毒感染人口腔鳞癌细胞系CAL-27建立FAK敲降细胞系和对照细胞系(FAK KD组和shRNA对照组),采用噻唑蓝(MTT)分析两组细胞的增殖能力;采用划痕实验和Transwell实验分析两组肿瘤细胞的迁移和侵袭水平;采用蛋白质印迹法(Western blot)分析两组细胞上皮-间充质转化水平。组间数据比较采用t检验。结果:癌旁组织FAK蛋白表达水平(154.23±23.09)明显低于肿瘤组织(351.29±30.18),差异有统计学意义(t=4.109,P<0.05)。对照组细胞吸光度(A)值(2.11±0.29)明显高于FAK KD组(1.52±0.26),差异有统计学意义(t=3.281,P<0.05)。对照组细胞克隆形成数量[(133.48±5.91)个]明显高于FAK KD组[(59.87±5.10)个],差异有统计学意义(t=4.289,P<0.05)。对照组细胞划痕愈合率[(85.62±6.08)%]明显高于FAK KD组[(53.18±4.87)%],差异有统计学意义(t=3.948,P<0.05)。对照组细胞迁移数量[(149.71±18.32)个]明显高于FAK KD组[(81.58±8.32)个],差异有统计学意义(t=4.770,P<0.05)。对照组细胞上皮细胞标志物E-钙黏蛋白(E-cadherin)表达水平(1.28±0.25)明显高于FAK KD组(0.63±0.19),差异有统计学意义(t=3.995,P<0.05)。对照组细胞间质细胞标志物N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)表达水平(0.84±0.18、0.96±0.20)明显低于FAK KD组(1.79±0.23、2.39±0.20),差异有统计学意义(t=3.027、4.318,P<0.05)。结论:FAK蛋白在口腔鳞癌组织中呈高表达,参与口腔鳞癌细胞的增殖、迁移、侵袭和上皮-间充质转化过程。

Objective To investigate the expression of focal adhesion kinase(FAK)in oral squamous cell carcinoma(OSCC)and its relationship with tumor cell migration and invasion.Methods Totally 79 cases of OSCC and adjacent tissues collected in the First Affiliated Hospital of Xinxiang Medical College and the Third Affiliated Hospital of Xinxiang Medical College from September 2015 to September 2018 were selected as research samples,and the expression level of FAK protein in adjacent tissues and OSCC tissues was analyzed by immunohistochemistry.FAK knockdown cell lines and control cell lines[FAK KD group and short hairpin RNA(shRNA)control group]were established by lentivirus infection of human OSCC cell line CAL-27 with FAK shRNA and control shRNA respectively.The proliferation in two groups was analyzed by methyl thiazolyl tetrazolium(MTT)assay.The migration and invasion of tumor cells in the two groups were analyzed by scratch test and Transwell.The level of epithelial mesenchymal transition(EMT)was analyzed by Western blotting.The data were compared between groups by t test.Results The expression level of FAK protein(154.23±23.09)was significantly lower than that in tumor tissue(351.29±30.18,t=4.109,P<0.05).The absorbance(A)value of control group(2.11±0.29)was significantly higher than that of FAK KD group(1.52±0.26,t=3.281,P<0.05).The number of cell clone formation in control group(133.48±5.91)was significantly greater than that in FAK KD group(59.87±5.10,t=4.289,P<0.05).The wound healing rate in the control group[(85.62±6.08)%]was significantly higher than that in the FAK KD group[(53.18±4.87)%,t=3.948,P<0.05].The number of migrating cells in the control group(149.71±18.32)was significantly greater than that in the FAK KD group(81.58±8.32,t=4.770,P<0.05).The expression level of E-cadherin in the control group(1.28±0.25)was significantly higher than that in the FAK KD group(0.63±0.19,t=3.995,P<0.05).The expression levels of N-cadherin and vimentin in the control group(0.84±0.18,0.96±0.20)weas significantly lower than those in the FAK KD group(1.79±0.23,2.39±0.20,t=3.027,4.318,P<0.05).Conclusion FAK protein is highly expressed in OSCC,which is involved in the proliferation,migration,invasion and EMT of OSCC.