| 杜雅冰,靳水玲,崔 抗.HOXC6、HOXC8基因对食管鳞癌细胞凋亡的影响及在接受手术切除食管鳞癌患者中的临床意义[J].中国肿瘤,2019,28(7):535-542. |
| HOXC6、HOXC8基因对食管鳞癌细胞凋亡的影响及在接受手术切除食管鳞癌患者中的临床意义 |
| Effect of HOXC6 and HOXC8 Gene Expression on Prognosis of Patients with Esophageal Squamous Cell Carcinoma and Its Mechanisms |
| 投稿时间：2018-11-10 |
| DOI：10.11735/j.issn.1004-0242.2019.07.A011 |
|
 |
| 中文关键词: 食管鳞癌 HOXC6 HOXC8 预后 凋亡 |
| 英文关键词:esophageal squamous cell carcinoma HOXC6 HOXC8 prognosis apoptosis |
| 基金项目:河南省科技厅科技攻关项目（172102310058） |
|
| 摘要点击次数: 1643 |
| 全文下载次数: 367 |
| 中文摘要: |
| 摘 要：［目的］通过靶向敲低HOXC6和HOXC8基因，研究HOXC6和HOXC8表达对食管鳞癌细胞增殖水平、克隆形成能力、细胞周期分布及细胞凋亡等生物学特性的影响，探讨HOXC6和HOXC8对食管鳞癌细胞凋亡的调控以及HOXC6和HOXC8表达在手术切除的食管鳞癌患者中的临床意义。［方法］收集145例接受手术切除的食管鳞癌患者的癌组织标本进行HOXC6和HOXC8表达免疫组化检测，并和患者的总生存期（overall survival，OS）进行相关性分析。细胞实验中，构建敲低HOXC6和HOXC8的慢病毒表达载体(pGLV3-shHOXC6和pGLV3-shHOXC8)，转染食管鳞癌细胞株ECA109，分别采用Real-time PCR和Western blot方法检测HOXC6和HOXC8基因的敲低效率。分别采用CCK8法和软琼脂克隆形成实验检测敲低HOXC6和HOXC8基因表达后对ECA109细胞增殖的影响。采用流式细胞仪检测敲低HOXC6和HOXC8基因表达后对ECA109细胞周期分布及细胞凋亡的影响。［结果］ HOXC6和HOXC8高表达和低表达患者的预后具有显著差异。HOXC6高表达和低表达患者的中位OS分别为38.5和81.3个月，差异具有显著的统计学意义（P=0.001）。HOXC8高表达和低表达患者的中位OS分别为34.6和114.4个月，差异同样具有显著的统计学意义（P<0.001）。另外，细胞实验表明：转染shRNA可显著下调ECA109细胞中HOXC6和HOXC8 mRNA和蛋白表达水平。CCK8结果显示干扰组（HOXC6 shRNA或HOXC8 shRNA）的细胞生存率较对照组明显下降（P<0.05）；克隆形成实验显示干扰组细胞克隆形成能力较对照组明显降低；流式细胞仪检测结果显示干扰组出现明显的G1细胞周期阻滞，且干扰组细胞凋亡率明显高于对照组。［结论］ HOXC6和HOXC8高表达对食管鳞癌患者预后有不良影响。敲低HOXC6和HOXC8基因可促进食管鳞癌细胞株ECA109凋亡。靶向调控HOXC6和HOXC8可能抑制食管鳞癌的发生与发展。 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the effects of HOXC6 and HOXC8 expression on prognosis of patients with esophageal squamous cell carcinoma (ESCC) and its mechanism. ［Methods］ One hundred and forty-five patients with ESCC undergoing surgical resection were enrolled in the study. The expression of HOXC6 and HOXC8 in ESCC cancer tissue was detected by immunohistochemistry method. And the association of HOXC6 and HOXC8 expression with overall survival (OS) of patients was analyzed. The lentiviral expression vectors pGLV3-shHOXC6 and pGLV3-shHOXC8 were constructed and transfected into esophageal squamous carcinoma cell line ECA109. Real-time RT-PCR and Western blot methods were employed to evaluate the transfection efficiency. CCK8 assay and soft agar colony formation assay were employed to detect the effects of HOXC6 and HOXC8 gene depletion on proliferation of ECA109 cells. Flow cytometry was employed to analyze cell cycle and apoptosis of ECA109 cells. ［Results］ The median OS of patients with high HOXC6 expression was significantly lower than that of patients with low expression patients (38.5 months vs 81.3 months，P=0.001). And the median OS of patients with high HOXC8 expression was significantly lower than hat of patients with low expression (34.6 months vs 114.4 months，P<0.001). Additionally，cell experiments indicated that transfection of shRNA significantly down-regulated HOXC6 and HOXC8 mRNA and protein expression in ECA109 cells. CCK8 assay showed that the cell proliferation rates of the interference groups (HOXC6 shRNA or HOXC8 shRNA) were significantly decreased，compared with control group (P<0.05). The clonogenic assay showed that the cell cloning ability of the interfering groups were also significantly decreased，compared with control group. Flow cytometry showed that the proportion in cell cycle arrest phase and the apoptosis rate in the interference group were significantly increased，compared with control group. ［Conclusion］ The high expression of HOXC6 or HOXC8 has an adverse influence on prognosis of patients with ESCC. HOXC6 and HOXC8 gene silencing can promote the apoptosis of ESCC ECA109 cells. The targeted regulation of HOXC6 and HOXC8 might inhibit the occurrence and development of esophageal squamous cell carcinoma. |
|
在线阅读
查看全文 查看/发表评论 下载PDF阅读器 |