过表达Beclin 1对食管癌Eca109细胞生物学行为的影响

Effect of Beclin 1 overexpression on biological behavior of esophageal cancer cell line Eca109

摘要:

目的：探讨自噬基因Beclin 1对食管癌细胞Eca109生物学行为的影响。方法：运用Beclin 1过表达质粒pIRES2-Zs－Green1-h Beclin 1转染食管癌 Eca109细胞株，RT-PCR和Western blot检测Beclin 1的mRNA和蛋白的表达；电子显微镜观察转染后自噬体的形成情况；MTT法检测转染前后各组细胞生长曲线的变化；血管形成实验检测其对Eca109细胞株血管形成能力的影响；Transwell小室实验检测其对Eca109细胞株迁移和侵袭能力影响；流式细胞技术检测目的基因组和对照组细胞周期的变化。结果：pIRES2-ZsGreen-hBeclin1质粒成功转染食管癌Eca109细胞株。目的基因组Eca109细胞较空载体组和未转染组生长明显抑制（P=0.000，P=0.000）；目的基因组Eca109细胞迁移、侵袭及血管形成能力明显低于对照组（P值均<0.05）；Beclin 1稳定表达后,S期细胞明显减少，细胞增殖受到抑制。结论：Beclin 1基因可以作为食管癌生长及转移的一种抑制基因。

Abstract:

Objective：To investigate the effect of Beclin 1 overexpression on the biological behavior of esophageal cancer cell line E－ca109. Methods：The recombinant eukaryotic expression vector pIRES2-ZsGreen-h Beclin 1 was transfected into Eca109 cell line. The mRNA and protein expressions of Beclin 1 were determined by RT-PCR and Western blot. Autophagosome was observed by electron microscopy. Changes in cell growth curve were determined by MTT assay and cell cycle was evaluated by flow cytometry. The effects of Beclin 1 overexpression on the migration and invasion of esophageal cancer cell line Eca109 were detected by Transwell chamber experiment. Vascular formation experiment in vitro was applied to determine the angiogenesis capability of Eca109. Results：The proliferation of Eca109 cells was obviously inhibited in Beclin 1 group than in empty vector group and control group. The cell in－vasion，angiogenesis and migration abilities were reduced Beclin 1 group than in control group（all P<0.05）. The proportion of cells at S stage was decreased significantly and the cell proliferation was inhibited obviously after steady expression of Beclin 1. Conclusion：Beclin 1 may be used as one of suppressor genes for esophageal cancer growth and metastasis.