下调黏蛋白1对人舌鳞状细胞癌Tca-8113细胞裸鼠皮下移植瘤生长的影响

Effect of Down-regulation of Mucin-1 on Growth of Xenografts of Human Tongue Squamous Cell Carcinoma Tca-8113 in Nude Mice

目的:探究下调黏蛋白1(mucin1,MUC1)表达对人舌鳞状细胞癌Tca-8113细胞裸鼠移植瘤生长的影响。方法:采用MUC1-短发卡RNA(short hairpin RNA,shRNA)慢病毒颗粒感染Tca-8113细胞,实时荧光定量聚合酶链反应(quantitative real-time PCR,qRT-PCR)和Western blot检测慢病毒感染后Tca-8113细胞中MUC1 mRNA及蛋白表达水平。皮下注射经慢病毒感染的Tca-8113细胞构建裸鼠移植瘤模型,观察肿瘤体积并绘制生长曲线。采用苏木精-伊红(hematoxylin-eosin,HE)染色和脱氧核糖核酸末端转移酶介导的缺口末端标记法(terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling,TUNEL)染色观察移植瘤组织病变及细胞凋亡情况;Western blot检测移植瘤组织中MUC1、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关蛋白(Bcl-2-associated X,Bax)、半胱氨酰天冬氨酸特异性蛋白酶-3(cysteine aspartic acid specific protease-3,Caspase-3)剪切体(Cleaved-caspase-3)和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)等蛋白表达水平。结果:慢病毒感染后Tca-8113细胞中MUC1 mRNA和蛋白表达水平显著降低(P<0.05)。与对照组及阴性对照组(MUC1-shCON)比较,MUC1-shRNA组裸鼠移植瘤体积明显减小(P<0.05),移植瘤组织细胞凋亡情况严重,且MUC1、Bcl-2和PCNA等蛋白表达水平明显降低(P<0.05),而Bax和Cleaved-caspase-3等蛋白表达水平升高(P<0.05)。结论: MUC1下调后可明显抑制人舌鳞状细胞癌Tca-8113细胞裸鼠皮下移植瘤生长。

Objective:To investigate the effect of down-regulation of mucin-1 (MUC1) expression on the growth of xenografts of human tongue squamous cell carcinoma Tca-8113 in nude mice.Methods:Tca-8113 cells were infected with MUC1-short hairpin RNA (shRNA) lentivirus.Quantitative real-time PCR (qPT-PCR) and western blot were used to detect the expression of MUC1 in infected Tca-8113 cells.The squamous cell carcinoma of tongue xenografts in nude mice were constructed by subcutaneous injection of Tca-8113 cells infected with MUC1-shRNA lentivirus.The volume of tumors was observed and the growth curve was drawn.Hematoxylin-eosin (HE) and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining were used to observe the pathological changes and apoptosis of xenografts.The expression of MUC1,B-cell lymphoma-2 (Bcl-2),Bcl-2-associated X (Bax),Cleaved-cysteine aspartic acid specific protease-3 (Cleaved-caspase-3),and proliferating cell nuclear antigen (PCNA) in tumors were detected by western blot.Results:After infected with MUC1-shRNA lentivirus,the mRNA and protein expression of MUC1 in Tca-8113 cells decreased significantly ( P <0.05).Compared with the control group and negative control group (MUC1-shCON),the volume of tumors in the MUC1-shRNA group was significantly reduced ( P <0.05),the apoptosis of transplanted tumor tissue was serious,and the expressions of MUC1,Bcl-2 and PCNA were significantly decreased (P<0.05),while the expressions of Bax and cleaves-caspase-3 protein were increased ( P < 0.05).Conclusion:The down-regulation of MUC1 can significantly inhibit the growth of tumor xenograft of human tongue squamous cell carcinoma Tca-8113 in nude mice.