Cáceres Blanca, Ramirez Alberto, Carrillo Esmeralda, Jimenez Gema, Griñán-Lisón Carmen, López-Ruiz Elena, Jiménez-Martínez Yaiza, Marchal Juan A, Boulaiz Houria
Motril Health Center, Hospital Santa Ana, Motril, 18600 Granada, Spain.
Biopathology and Medicine Regenerative Institute (IBIMER), University of Granada, 18016 Granada, Spain.
Cancers (Basel). 2019 Feb 23;11(2):264. doi: 10.3390/cancers11020264.
Despite the great advances in cancer treatment, colorectal cancer has emerged as the second highest cause of death from cancer worldwide. For this type of tumor, the use of suicide gene therapy could represent a novel therapy. We recently demonstrated that co-expression of and dramatically inhibits proliferation of the DLD-1 colon cell line. In the present manuscript, we try to establish the mechanism underlying the enhanced induction of apoptosis by triggering both and expression in colon tumor cells. Scanning microscopy reveals that simultaneous expression of and induces the apparition of many "pores" in the cytoplasmic membrane not detected in control cell lines. The formation of pores induced by the gene and accentuated by apoptin results in cell death by necrosis. Moreover, we observed the presence of apoptotic cells. Performing protein expression analysis using western blot, we revealed an activation of mitochondrial apoptosis (increased expression of Pp53, cytochrome c, Bax, and caspase 9) and also the involvement of the extrinsic pathway through caspase 8activation. In conclusion, in this manuscript we demonstrate for the first time that the extrinsic pathway of apoptosis and pore formation is also involved in the cell death caused by the co-expression of the and genes. Our results suggest that co-expression of and genes induces an increase in post-apoptotic necrotic cell death and could be a valuable tool in the design of new antitumor strategies focused on the enhancement of the immune response against cancer cell death.
尽管癌症治疗取得了巨大进展，但结直肠癌已成为全球癌症死亡的第二大原因。对于这种类型的肿瘤，自杀基因疗法可能代表一种新型疗法。我们最近证明，[具体基因名称1]和[具体基因名称2]的共表达显著抑制了DLD-1结肠癌细胞系的增殖。在本论文中，我们试图确定通过在结肠肿瘤细胞中触发[具体基因名称1]和[具体基因名称2]的表达来增强凋亡诱导的潜在机制。扫描显微镜显示，[具体基因名称1]和[具体基因名称2]的同时表达诱导了细胞质膜上许多“孔”的出现，而在对照细胞系中未检测到。由[具体基因名称1]基因诱导并由凋亡素加剧的孔的形成导致细胞坏死性死亡。此外，我们观察到了凋亡细胞的存在。通过蛋白质印迹法进行蛋白质表达分析，我们发现线粒体凋亡被激活（Pp-53、细胞色素c、Bax和半胱天冬酶9的表达增加），并且通过半胱天冬酶8的激活也涉及外源性途径。总之，在本论文中，我们首次证明凋亡的外源性途径和孔的形成也参与了由[具体基因名称1]和[具体基因名称2]基因共表达引起的细胞死亡。我们的结果表明，[具体基因名称1]和[具体基因名称]基因的共表达诱导了凋亡后坏死性细胞死亡的增加，并且可能是设计针对增强针对癌细胞死亡的免疫反应的新抗肿瘤策略的有价值工具。
