肿瘤抑制基因p53沉默人肺腺癌A549细胞模型的建立及功能初步研究

Establishment of a human lung adenocarcinoma A549 cell model knockdown by tumor suppressor gene p53 and its functional study

目的 通过慢病毒介导构建肿瘤抑制基因p53沉默人肺腺癌A549细胞模型,检测p53沉默后对人肺腺癌A549细胞自噬反应的影响。方法 利用pCDH-MSCV-MCS-EF1-Puro慢病毒载体构建重组质粒,将重组质粒和空载质粒转染293T细胞,收集病毒液后,感染A549细胞,构建沉默细胞模型。将空载质粒组和重组质粒组分别设置为对照组和实验组。利用Western blotting技术检测p53沉默效率和自噬相关指标p62和LC3。结果 Western blotting检测p53沉默人肺腺癌A549细胞模型构建成功;Western blotting结果显示,p53沉默下调p62和上调LC3,促进细胞自噬。结论 成功构建慢病毒介导的肿瘤抑制基因p53沉默人肺腺癌A549细胞模型,该细胞模型能够促进自噬的发生发展,可能为临床治疗肺癌疾病提供一种新的思路。

Objective To establish a model of human lung adenocarcinoma A549 cells silenced by tumor suppressor gene p53 mediated by lentivirus, and to investigate the effect of p53 silencing on autophagy in human lung adenocarcinoma A549 cells. Methods The recombinant plasmid was constructed by pcdh-mscv-mcs-ef1-puro lentivirus vector. The recombinant plasmid and empty plasmid were transfected into 293 T cells. After collecting the viral solution, A549 cells were infected to construct the silence cell model. The empty plasmid group and recombinant plasmid group were set as control group and experimental group respectively. Western blot was used to detect p53 silencing efficiency and autophagy related indicators p62 and LC3. Results Western blot analysis showed that the model of p53 silencing human lung adenocarcinoma A549 cells was successfully constructed, and p53 down-regulated p62 and up-regulated LC3, which promoted autophagy. Conclusion We successfully construct a lentivirus mediated tumor suppressor gene p53 silencing human lung adenocarcinoma A549 cell model, and the cell model can promote the occurrence and development of autophagy, which provides a new idea for clinical treatment of lung cancer.