口腔医学研究 ›› 2019, Vol. 35 ›› Issue (8): 776-780.DOI: 10.13701/j.cnki.kqyxyj.2019.08.013

• 口腔颌面外科学研究 • 上一篇    下一篇

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

邓轩1, 阳芳2*, 唐西清1   

  1. 1. 南华大学附属第二医院口腔科 湖南 衡阳 421001;
    2. 南华大学附属第二医院神经外科 湖南 衡阳 421001
  • 收稿日期:2018-12-31 出版日期:2019-08-28 发布日期:2019-08-22
  • 通讯作者: 阳芳,E-mail:mydy537@163.com
  • 作者简介:邓轩(1985~),男,湖北荆州人,硕士,主治医师,主要从事口腔疾病诊疗工作。

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

DENG Xuan1, YANG Fang2*, TANG Xiqing1   

  1. 1. Department of Stomatology, The Second Hospital University of South China, Hengyang 421001, China;
    2. Department of Neurosurgery, The Second Hospital University of South China, Hengyang 421001, China
  • Received:2018-12-31 Online:2019-08-28 Published:2019-08-22

摘要: 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。

关键词: 黏蛋白1, 舌鳞状细胞癌, 移植瘤, 凋亡

Abstract: 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.

Key words: Mucin-1, tongue squamous cell carcinoma, xenografts, apoptosis