沉默PIWIL2抑制口腔鳞癌细胞CAL27增殖、侵袭、迁移和裸鼠移植瘤生长  

Silencing PIWIL2 on inhibiting the proliferation,invasion and migration of oral squamous cell carcinoma cells CAL27 and the growth of transplanted tumors in nude mice

作　　者：郭谊 韩炎 周丽芝[3] 王俊华 马跃 鲁大鹏[5] GUO Yi;HAN Yan;ZHOU Li-zhi;WANG Jun-hua;MA Yue;LU Da-peng(Department of Stomatology,Tangshan Hospital of Traditional Chinese Medicine,Tangshan Hebei 063000,China;Department of Stomatology,Wen’an County Hospital,Langfang Hebei 065800,China;Department of Stomatology,Tangshan Eighth Hospital,Tangshan Hebei 063000,China;Department of Cardiology,Wen’an County Hospital,Langfang Hebei 065800,China;Center of Emergency Comprehensive Treatment,Beijing Stomatological Hospital Affiliated to Capital Medical University,Beijing 100050,China)

机构地区：[1]唐山市中医医院口腔科,河北唐山063000 [2]文安县医院口腔科,河北廊坊065800 [3]唐山市第八医院口腔科,河北唐山063000 [4]文安县医院心内科,河北廊坊065800 [5]首都医科大学附属北京口腔医院急诊综合治疗中心,北京100050

出　　处：《局解手术学杂志》2022年第10期857-862,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：廊坊市科技支撑计划项目(2018013150)。

摘　　要：目的探究精原干细胞自我更新基因PIWIL2对口腔鳞癌细胞CAL27增殖、侵袭、迁移和裸鼠移植瘤生长的影响。方法将口腔鳞癌细胞CAL27随机分为空白组(不作处理)、阴性对照组(转染慢病毒阴性对照病毒scramble)和PIWIL2-shRNA慢病毒组(转染PIWIL2-shRNA慢病毒)。采用实时荧光定量PCR和蛋白印迹法检测细胞中PIWIL2 mRNA及蛋白表达水平,集落形成实验检测细胞增殖能力,Transwell实验和划痕实验检测细胞侵袭和迁移能力,JC-1探针检测线粒体膜电位变化,蛋白印迹法检测Ki67、血管内皮生长因子(VEGF)、E-cadherin、N-cadherin、Snail表达水平。裸鼠皮下注射慢病毒感染的CAL27细胞构建裸鼠移植瘤模型,免疫组化染色检测肿瘤组织中VEGF的表达水平,观察沉默PIWIL2对肿瘤生长的影响。结果PIWIL2-shRNA慢病毒感染后CAL27细胞中PIWIL2 mRNA及蛋白表达水平显著降低(P<0.05)。与阴性对照组比较,PIWIL2-shRNA慢病毒组细胞集落形成率,侵袭细胞数,划痕愈合率,细胞中Ki67、VEGF、N-cadherin、Snail蛋白表达水平,移植瘤重量、体积,移植瘤组织中VEGF阳性表达率显著降低/减少,细胞中E-cadherin蛋白表达水平显著升高,差异均有统计学意义(P<0.05)。JC-1探针检测结果显示,PIWIL2-shRNA慢病毒感染后细胞中红色荧光减少,绿色荧光增多。结论沉默PIWIL2可明显抑制口腔鳞癌细胞CAL27增殖、侵袭、迁移及裸鼠移植瘤的生长。Objective To explore the effects of spermatogonial stem cells self-renewal gene PIWIL2 on the proliferation,invasion and migration of oral squamous cell carcinoma cells CAL27 and the growth of transplanted tumors in nude mice.Methods Oral squamous cell carcinoma cells CAL27 were randomly divided into the blank group(without any treatment),the negative control group(transfected with lentivirus negative control virus scramble)and the PIWIL2-shRNA lentivirus group(transfected with PIWIL2-shRNA lentivirus).The expression levels of PIWIL2 mRNA and protein in cells were detected by real-time fluorescent quantitative PCR and Western blot,cells proliferation was detected by colony formation assay,cells invasion and migration were detected by Transwell assay and scratch wound healing assay,and change of mitochondrial membrane potentials was detected by JC-1 probe.The expression levels of Ki67,vascular endothelial growth factor(VEGF),E-cadherin,N-cadherin and Snail proteins were detected by Western blot.The transplanted tumor model of nude mice was constructed by subcutaneous injection of CAL27 cells infected with lentivirus in nude mices,the expression level of VEGF in tumor tissues was detected by immunohistochemistry,and the effect of silencing PIWIL2 on the growth of tumors was observed.Results After infected with PIWIL2-shRNA lentivirus,expression levels of PIWIL2 mRNA and protein in CAL27 cells were significantly decreased(P<0.05).Compared with the negative control group,the rate of cells colony formation,the number of invasion cells,the scratch wound healing rate,the expression levels of Ki67,VEGF,N-cadherin and Snail proteins in cells,the weight and volume of transplanted tumors,and the positive expression rate of VEGF in transplanted tumor tissues were significantly decreased in the PIWIL2-shRNA lentivirus group,while the expression level of E-cadherin protein in cells was significantly increased,with statistically significant differences(P<0.05).The test results of JC-1 probe showed that red fluorescence in cells

关 键 词：口腔鳞癌 PIWIL2 增殖 侵袭 迁移 移植瘤 

分 类 号：R739.8[医药卫生—肿瘤]