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自噬是一个进化上保守的溶酶体降解途径,激活自噬可以降解有缺陷的细胞器而发挥抑制肿瘤的作用,Beclin 1是关键的自噬和肿瘤抑制基因。该研究检测了Beclin 1蛋白在食管癌组织中的表达情况,构建Beclin 1的重组质粒并转染人食管癌Eca109细胞株。分别以电子显微镜观察自噬体的形成和Eca109细胞的自噬性死亡情况,RT-PCR检测Beclin 1基因的mRNA表达,Westernblot法检测Beclin 1及Bcl-2、P53的表达水平,裸鼠成瘤实验检测Beclin 1基因对Eca109细胞体内增殖的影响。结果显示,Beclin 1蛋白在食管癌组织中相对于癌旁正常食管组织低表达(P<0.05);成功构建了Beclin 1重组质粒并将其转染到Eca109细胞中,电子显微镜观察到自噬体形成,RT-PCR和Western blot显示Beclin 1基因过表达。升高Beclin 1蛋白的表达水平能降低Bcl-2表达和上调P53蛋白,使Eca109细胞的体内致瘤性减弱。过表达Beclin 1基因对食管癌有很好的抑制作用,这将为食管癌的靶向治疗奠定基础。
Autophagy is an evolutionarily conserved lysosomal degradation pathway. Activation of autophagy can degrade defective organelles and play a tumor suppressor role. Beclin 1 is a key autophagy and tumor suppressor gene. The study examined the expression of Beclin 1 protein in esophageal cancer tissues, constructed a recombinant plasmid of Beclin 1 and transfected into human esophageal cancer Eca109 cell line. The formation of autophagosome and the autophagic death of Eca109 cells were observed by electron microscope. The mRNA expression of Beclin 1 gene was detected by RT-PCR. The expression of Beclin 1, Bcl-2 and P53 was detected by Western blot. Effect of Beclin 1 gene on the proliferation of Eca109 cells in vivo. The results showed that Beclin 1 protein was overexpressed in esophageal cancer tissues compared with normal esophageal cancer tissues (P <0.05). Beclin 1 recombinant plasmid was successfully constructed and transfected into Eca109 cells. Autophagy was observed by electron microscopy Beclin 1 gene was overexpressed by RT-PCR and Western blot. Elevated Beclin 1 protein expression can decrease Bcl-2 expression and up-regulate P53 protein, resulting in the decrease of in vivo tumorigenicity of Eca109 cells. Overexpression of Beclin 1 gene has a good inhibitory effect on esophageal cancer, which will lay the foundation for the targeted therapy of esophageal cancer.