| 摘要: |
| 目的 通过腺相关病毒介导的小干扰RNA(siRNA)抑制人食管鳞癌细胞Eca109中ΔNp63α的表达,观察其对食管鳞癌细胞生物学特性的影响。方法 构建H1启动子驱动的表达针对ΔNp63α的siRNA重组腺相关病毒AAV-ΔNp63αshRNA并感染Eca109细胞。同时以感染空腺相关病毒AAV-Null的细胞及不加腺相关病毒的细胞分别作为对照组和空白对照组,观察siRNA干扰ΔNp63α表达在体外及体内对Eca109细胞生长、增殖、致瘤及凋亡的影响。结果 与对照组和空白对照组比较,Eca109细胞感染AAV-ΔNp63αshRNA后ΔNp63α蛋白表达下降(P均<0.05),细胞生长速度减慢(P均<0.05),细胞增殖指数 (proliferation index,PI) 降低(P均<0.01),裸鼠瘤体质量减轻(P均<0.05),细胞凋亡增加(P均<0.05)。结论 腺相关病毒介导的siRNA干扰能显著抑制Eca109细胞ΔNp63α的表达,减慢肿瘤细胞的增殖,促进肿瘤细胞凋亡,抑制肿瘤生长。 |
| 关键词: ΔNp63α 食管肿瘤 鳞状细胞肿瘤 Eca109细胞株 腺相关病毒 RNA干扰 |
| DOI： |
| 投稿时间：2013-04-24修订日期：2013-06-20 |
| 基金项目:国家自然科学基金(81201780),上海市科委医学引导类科技项目(114119a9300). |
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| Effect of small interfering RNA targeting ΔNp63α gene on the biological characteristics of human esophageal squamous carcinoma cells |
| LI Bai-ling1,DU Da-hai2,ZHANG Guan-xin1,XU Zhi-yun1* |
(1. Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
2. Department of Laboratory Diagnosis, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
*Corresponding author.) |
| Abstract: |
| Objective To construct a small interfering RNA (siRNA) vector targeting ΔNp63α and investigate ΔNp63α gene interference on the proliferation and apoptosis of human esophageal squamous carcinoma Eca109 cell line. Methods Adeno-associated virus (AAV)-ΔNp63αshRNA driven by H1 promoter was constructed and was used to infect Eca109 cells. AAV-Null and normal cell lines were utilized in the control group and blank control group, respectively. The influence of siRNA interference of ΔNp63α expression on the growth, proliferation, tumorigenic efficiency and apoptosis of Eca109 cells were analyzed in vitro and in vivo. Results Compared with the two control groups, the specific siRNA targeting ΔNp63α gene significantly down-regulated the expression of ΔNp63α protein levels in Eca109 cells (all P<0.05).The growth of Eca109 cells infected with AAV-ΔNp63αshRNA was significantly lower than those in the two control groups (all P<0.05). Cell cycle analysis showed the proliferation index (PI) of AAV-ΔNp63αshRNA infected cell line was significantly lower compared with the two control groups (all P<0.01). In vivo experiment exhibited that AAV-ΔNp63αshRNA infected cells resulted in a lower tumor weight in nude mice compared with the cells in the two control groups (all P<0.05). In addition, the apoptosis index (AI) of AAV-ΔNp63αshRNA infected cells were significantly higher than those of the other cell lines (P<0.05). Conclusion AAV-mediated expression of siRNA can significantly reduce ΔNp63α expression in Eca109 cells, slowing down the proliferation, promoting the apoptosis, and subsequently inhibiting the growth of tumor. |
| Key words: ΔNp63α esophageal neoplasms squamous cell neoplasms Eca109 cell line adeno-associated virus RNA interference |
