Wei Dongping, Zhang Qiang, Schreiber Jason S, Parsels Leslie A, Abulwerdi Fardokht A, Kausar Tasneem, Lawrence Theodore S, Sun Yi, Nikolovska-Coleska Zaneta, Morgan Meredith A
Department of Radiation Oncology, University of Michigan Medical School.
Department of Pharmacology, University of Michigan Medical School.
Transl Oncol. 2015 Feb;8(1):47-54. doi: 10.1016/j.tranon.2014.12.004.
In order to identify targets whose inhibition may enhance the efficacy of chemoradiation in pancreatic cancer, we previously conducted an RNAi library screen of 8,800 genes. We identified Mcl-1 (myeloid cell leukemia-1), an anti-apoptotic member of the Bcl-2 family, as a target for sensitizing pancreatic cancer cells to chemoradiation. In the present study we investigated Mcl-1 inhibition by either genetic or pharmacological approaches as a radiosensitizing strategy in pancreatic cancer cells. Mcl-1 depletion by siRNA produced significant radiosensitization in BxPC-3 and Panc-1 cells in association with Caspase-3 activation and PARP cleavage, but only minimal radiosensitization in MiaPaCa-2 cells. We next tested the ability of the recently identified, selective, small molecule inhibitor of Mcl-1, UMI77, to radiosensitize in pancreatic cancer cells. UMI77 caused dissociation of Mcl-1 from the pro-apoptotic protein Bak and produced significant radiosensitization in BxPC-3 and Panc-1 cells, but minimal radiosensitization in MiaPaCa-2 cells. Radiosensitization by UMI77 was associated with Caspase-3 activation and PARP cleavage. Importantly, UMI77 did not radiosensitize normal small intestinal cells. In contrast, ABT-737, an established inhibitor of Bcl-2, Bcl-XL, and Bcl-w, failed to radiosensitize pancreatic cancer cells suggesting the unique importance of Mcl-1 relative to other Bcl-2 family members to radiation survival in pancreatic cancer cells. Taken together, these results validate Mcl-1 as a target for radiosensitization of pancreatic cancer cells and demonstrate the ability of small molecules which bind the canonical BH3 groove of Mcl-1, causing displacement of Mcl-1 from Bak, to selectively radiosensitize pancreatic cancer cells.
为了鉴定抑制哪些靶点可增强胰腺癌放化疗的疗效,我们之前对8800个基因进行了RNA干扰文库筛选。我们鉴定出Bcl-2家族的抗凋亡成员Mcl-1(髓样细胞白血病-1)是使胰腺癌细胞对放化疗敏感的一个靶点。在本研究中,我们研究了通过基因或药理学方法抑制Mcl-1作为胰腺癌细胞放射增敏策略的效果。用小干扰RNA敲低Mcl-1可使BxPC-3和Panc-1细胞产生显著的放射增敏作用,伴有半胱天冬酶-3激活和聚ADP核糖聚合酶裂解,但在MiaPaCa-2细胞中仅产生极小的放射增敏作用。接下来,我们测试了最近鉴定出的Mcl-1选择性小分子抑制剂UMI77使胰腺癌细胞放射增敏的能力。UMI77导致Mcl-1与促凋亡蛋白Bak解离,并在BxPC-3和Panc-1细胞中产生显著的放射增敏作用,但在MiaPaCa-2细胞中产生极小的放射增敏作用。UMI77的放射增敏作用与半胱天冬酶-3激活和聚ADP核糖聚合酶裂解有关。重要的是,UMI77不会使正常小肠细胞放射增敏。相比之下,已证实的Bcl-2、Bcl-XL和Bcl-w抑制剂ABT-737未能使胰腺癌细胞放射增敏,这表明相对于其他Bcl-2家族成员,Mcl-1对胰腺癌细胞的辐射存活具有独特的重要性。综上所述,这些结果证实Mcl-1是胰腺癌细胞放射增敏的一个靶点,并证明了结合Mcl-1经典BH3凹槽、导致Mcl-1与Bak解离的小分子能够选择性地使胰腺癌细胞放射增敏。