Department of Urology, the First Affiliated Hospital of Bengbu Medical College, Bengbu, China.
Department of Urology, the Second Affiliated Hospital of Soochow University, Suzhou, China.
Neoplasma. 2020 Sep;67(5):1074-1084. doi: 10.4149/neo_2020_191115N01171. Epub 2020 Jun 2.
Death inducer obliterator (DIDO) is involved in apoptosis and embryonic stem cell self-renewal. Here, we investigate the effect of DIDO1 on bladder cancer cells and clarify the underlying molecular mechanism. Bladder cancer tissues and cell lines (T24, ScaBER, 5637), as well as normal bladder epithelial cells (SV-HUC-1), were used to measure the levels of DIDO1 mRNA and protein by qRT-PCR and western blot, respectively. The results indicated that DIDO1 was highly expressed in bladder cancer tissues and cell lines. And the expression of DIDO1 in T24 and 5637 cells was higher than that in ScaBER and SV-HUC-1 cells. The expression of DIDO1 was knocked down in T24 and 5637 cells by infection with shDIDO1-1 and shDIDO1-2 lentivirus. The growth of T24 and 5637 cells was monitored using Celigo, MTT assays, and colony formation assay. Apoptosis was examined by flow cytometric analysis. The effect of DIDO1 knockdown on tumorigenesis of T24 xenograft tumors was determined in nude mice. Reduction of DIDO1 mRNA resulted in reduced proliferation, decreased cell colony formation, increased apoptosis in vitro, and inhibited tumorigenesis in vivo. Furthermore, we identified signaling molecules involved in stress and apoptosis using the PathScan Antibody Array Kit and western blot. The depletion of DIDO1 significantly decreased the levels of phosphorylated SAPK/JNK, and Chk1/2, as well as the upregulating cleaved Caspase-7 expression. These results indicated that the potential mechanism of DIDO1 action might involve SAPK/JNK signaling cascades.
死亡诱导破坏因子（DIDO）参与细胞凋亡和胚胎干细胞自我更新。在此，我们研究了 DIDO1 对膀胱癌细胞的影响，并阐明了其潜在的分子机制。我们使用膀胱癌组织和细胞系（T24、ScaBER、5637）以及正常膀胱上皮细胞（SV-HUC-1），通过 qRT-PCR 和 Western blot 分别检测 DIDO1 mRNA 和蛋白的水平。结果表明，DIDO1 在膀胱癌组织和细胞系中高表达。并且，在 T24 和 5637 细胞中 DIDO1 的表达高于 ScaBER 和 SV-HUC-1 细胞。通过感染 shDIDO1-1 和 shDIDO1-2 慢病毒下调 T24 和 5637 细胞中的 DIDO1 表达。使用 Celigo、MTT 分析和集落形成分析监测 T24 和 5637 细胞的生长。通过流式细胞术分析检测细胞凋亡。在裸鼠中确定 DIDO1 敲低对 T24 异种移植肿瘤发生的影响。DIDO1 mRNA 的减少导致体外增殖减少、细胞集落形成减少、凋亡增加，并抑制体内肿瘤发生。此外，我们使用 PathScan 抗体阵列试剂盒和 Western blot 鉴定了参与应激和细胞凋亡的信号分子。DIDO1 的耗竭显著降低了磷酸化 SAPK/JNK 和 Chk1/2 的水平，以及上调的 cleaved Caspase-7 表达。这些结果表明，DIDO1 作用的潜在机制可能涉及 SAPK/JNK 信号级联。
