A Novel Combination RNAi toward Warburg Effect by Replacement with miR-145 and Silencing of PTBP1 Induces Apoptotic Cell Death in Bladder Cancer Cells.
A Novel Combination RNAi toward Warburg Effect by Replacement with miR-145 and Silencing of PTBP1 Induces Apoptotic Cell Death in Bladder Cancer Cells.
复制标题
DOI：
10.3390/ijms18010179
复制
发表时间：
2017-01-17
影响因子：
5.6
通讯作者：
Azuma H
中科院分区：
文献类型：
--
作者：
Takai T;Yoshikawa Y;Inamoto T;Minami K;Taniguchi K;Sugito N;Kuranaga Y;Shinohara H;Kumazaki M;Tsujino T;Takahara K;Ito Y;Akao Y;Azuma H
膀胱癌是最难控制的恶性肿瘤之一。我们探索了一种新的RNA干扰方法的使用驱动癌基因调节癌症特定的能量代谢的组合治疗的小干扰RNA（siRNA）和microRNA。转染miR-145和/或siR-PTBP 1的T24和253 JB-V细胞后，我们通过台盼蓝染料排斥试验、Western blot、Hoechst 33342染色、逆转录聚合酶链反应（RT-PCR）和电子显微镜检查细胞生长和基因表达的影响。然后评估具有miR-145和/或siR-PTBP 1的异种移植模型小鼠的抗癌效果。联合处理诱导更深和更长的生长抑制，并降低c-Myc和多聚嘧啶片段结合蛋白1（PTBP 1）的mRNA和蛋白质表达水平比每个单一处理。值得注意的是，联合治疗不仅通过抑制c-Myc/PTBP 1/PKMs轴损害了癌症特异性能量代谢，而且在体外和体内检查了MAPK/ERK和PI 3 K/AKT通路的失活。此外，联合治疗诱导细胞凋亡或自噬;但是，在一些细胞中，凋亡性细胞死亡伴随着自噬，因为染色质的浓缩和许多自噬体共存。这种联合治疗可能是一种新的RNA干扰策略，通过系统性沉默膀胱癌细胞中的瓦尔堡效应促进驱动癌基因PTBP 1。
Bladder cancer is one of the most difficult malignancies to control. We explored the use of a novel RNA-interference method for a driver oncogene regulating cancer specific energy metabolism by the combination treatment with a small interfering RNA (siRNA) and a microRNA. After transfection of T24 and 253JB-V cells with miR-145 and/or siR-PTBP1, we examined the effects of cell growth and gene expression by performing the trypan blue dye exclusion test, Western blot, Hoechst 33342 staining, reverse transcription polymerase chain reaction (RT-PCR), and electron microscopy. The anti-cancer effects of xenograft model mice with miR-145 and/or siR-PTBP1 were then assessed. The combination treatment induced the deeper and longer growth inhibition and reduced the levels of both mRNA and protein expression of c-Myc and polypyrimidine tract-binding protein 1 (PTBP1) more than each single treatment. Notably, the combination treatment not only impaired the cancer specific energy metabolism by inhibiting c-Myc/PTBP1/PKMs axis but also inactivated MAPK/ERK and PI3K/AKT pathways examined in vitro and in vivo. Furthermore, the combination treatment induced apoptosis or autophagy; but, in some cells, apoptotic cell death was accompanied by autophagy, because the condensation of chromatin and many autophagosomes were coexistent. This combination treatment could be a novel RNA-interference strategy through the systemic silencing of the Warburg effect-promoting driver oncogene PTBP1 in bladder cancer cells.