Tai Jun, Li Ai-Dong, Rao Yuan-Sheng, Huang Yu-Bei, Huang Zhi-Gang, Yu Zhen-Kun, Chen Xiao-Hong, Zhou Wei-Guo, Xiao Xiao, Wang Shen, Han Yang, Liu Qiao-Yin, Fang Ju-Gao, Ni Xin
Beijing Key Laboratory for Pediatric Otolaryngology, Head and Neck Science, Beijing Pediatric Research Institute,Beijing Children's Hospital, Capital Medical University, Beijing 100045, China.
Department of Otorhinolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100029, China. Email:
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2013 Sep;48(9):761-4.
The effects of lentivirus-mediated suppression of Cyclin Y (CCNY) expression on the proliferation of laryngeal cancer cells were investigated in vitro.
The lentivirus vectors containing a small hairpin RNA (shRNA) to target CCNY were constructed.Hep-2 cells were divided into the following two experimental groups:the negative control group (control lentivirus infected cells) and CCNY knockdown group (CCNY shRNA-expressing lentivirus infected cells). After Hep-2 cells were infected, Real-time PCR was used to measure CCNY expression. The influence of CCNY on the proliferation of laryngeal cancer cells were assessed using MTT and colony formation experiments.Each experiment was performed in triplicate and repeated three times.
Lentiviruses expressing shRNA against CCNY were constructed and Hep-2 cells were infected with above mentioned lentivirus at MOI (Multiplicity of infection) of 120.Real-time PCR analysis showed that the mRNA expression of CCNY in Hep-2 cells in the knockdown group was significantly decreased (P < 0.05); the mRNA level of CCNY was 75.3% lower in the si-CCNY group than in the si-CTRL group. After 5 days of lentiviral infection, the cell viability was significantly lower in cells infected with the CCNY-shRNA lentivirus compared to cells infected with the control lentivirus following a 6-day incubation. The colony number was decreased by 60% in Hep-2 cells infected with the CCNY-shRNA-lentivirus infected cells following a 10-day incubation.
The results suggested that lentivirus-mediated downregulation of CCNY expression decreased the proliferation and growth potency of laryngeal cancer cells.Lentiviruses delivering shRNA against CCNY may be a promising tool for laryngeal cancer therapy.
在体外研究慢病毒介导的细胞周期蛋白Y(CCNY)表达抑制对喉癌细胞增殖的影响。
构建针对CCNY的短发夹RNA(shRNA)慢病毒载体。将Hep-2细胞分为以下两个实验组:阴性对照组(对照慢病毒感染细胞)和CCNY敲低组(表达CCNY shRNA的慢病毒感染细胞)。Hep-2细胞感染后,采用实时荧光定量PCR检测CCNY表达。使用MTT和集落形成实验评估CCNY对喉癌细胞增殖的影响。每个实验重复三次,共进行三次。
构建了表达针对CCNY的shRNA的慢病毒,并以感染复数(MOI)为120用上述慢病毒感染Hep-2细胞。实时荧光定量PCR分析显示,敲低组Hep-2细胞中CCNY的mRNA表达显著降低(P<0.05);si-CCNY组的CCNY mRNA水平比si-CTRL组低75.3%。慢病毒感染5天后,与感染对照慢病毒的细胞在孵育6天后相比,感染CCNY-shRNA慢病毒的细胞活力显著降低。孵育10天后,感染CCNY-shRNA慢病毒的Hep-2细胞集落数减少了60%。
结果表明,慢病毒介导的CCNY表达下调降低了喉癌细胞的增殖和生长能力。递送针对CCNY的shRNA的慢病毒可能是一种有前景的喉癌治疗工具。