Inhibition of NANOG/NANOGP8 downregulates MCL-1 in colorectal cancer cells and enhances the therapeutic efficacy of BH3 mimetics.
Inhibition of NANOG/NANOGP8 downregulates MCL-1 in colorectal cancer cells and enhances the therapeutic efficacy of BH3 mimetics.
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DOI：
10.1158/1078-0432.ccr-14-1134
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发表时间：
2014-11-01
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影响因子：
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通讯作者：
Jessup JM
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文献类型：
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作者：
Mattoo AR;Zhang J;Espinoza LA;Jessup JM
大肠癌中高水平的BCL-2家族成员导致对治疗的抵抗。NANOG或其副产物NANOGP 8的抑制降低结肠直肠癌细胞的增殖、干性和致瘤性。我们的假设是，NANOG/NANOGP 8的抑制通过降低MCL-1（一种促生存BCL-2蛋白）的表达，增强了靶向BCL-2家族成员的BH 3模拟物在结直肠癌细胞中的细胞毒性作用。将慢病毒载体（LV）shRNA导入NANOG（shNG-1）或NANOGP 8（shNp 8 -1）转导的结直肠癌细胞，所述结直肠癌细胞也在结直肠癌异种移植物中体内和体外暴露于BH 3模拟物ABT-737或ABT-199，其中测量增殖、蛋白质和基因表达以及凋亡。克隆A和CX-1对ABT-737和ABT-199敏感，IC_（50）为2 ~ 9 μmol/L，而LS 174 T对IC_（50）为18 ~ 30 μmol/L。抗性与LS 174 T中的高MCL-1表达相关。与单独使用BH 3模拟物的作用相比，LVshNG-1或LVshNp 8 -1降低了ABT-737或ABT-199处理的结直肠癌细胞中MCL-1的表达，增加了凋亡，并降低了再铺板效率。单独抑制或过表达MCL-1复制了LVshNG-1或LVshNp 8 -1在增加或减少BH 3模拟物引起的细胞凋亡中的作用。与未处理的对照或仅用LV shRNA或ABT-737处理相比，联合治疗抑制了体内LS 174 T异种移植物的生长。NANOGP 8或NANOG的抑制增强靶向BCL-2家族成员的BH 3模拟物的细胞毒性。靶向NANOG的基因治疗可能会增加BH 3模拟物在结直肠癌中的疗效。
High levels of BCL-2 family members in colorectal carcinoma cause resistance to treatment. Inhibition of NANOG or its paralog NANOGP8 reduces the proliferation, stemness, and tumorigenicity of colorectal carcinoma cells. Our hypothesis was that inhibition of NANOG/NANOGP8 enhances the cytotoxic effect of BH3 mimetics targeting BCL-2 family members in colorectal carcinoma cells through reducing expression of MCL-1, a prosurvival BCL-2 protein. Lentiviral vector (LV) shRNA to NANOG (shNG-1) or NANOGP8 (shNp8-1) transduced colorectal carcinoma cells that were also exposed to the BH3 mimetics ABT-737 or ABT-199 in vivo in colorectal carcinoma xenografts and in vitro where proliferation, protein and gene expression, and apoptosis were measured. Clone A and CX-1 were sensitive to ABT-737 and ABT-199 at IC50s of 2 to 9 μmol/L but LS174T was resistant with IC50s of 18 to 30 μmol/L. Resistance was associated with high MCL-1 expression in LS174T. LVshNG-1 or LVshNp8-1 decreased MCL-1 expression, increased apoptosis, and decreased replating efficiency in colorectal carcinoma cells treated with either ABT-737 or ABT-199 compared with the effects of either BH3 mimetic alone. Inhibition or overexpression of MCL-1 alone replicated the effects of LVshNG-1 or LVshNp8-1 in increasing or decreasing the apoptosis caused with the BH3 mimetic. The combination therapy inhibited the growth of LS174T xenografts in vivo compared with untreated controls or treatment with only LV shRNA or ABT-737. Inhibition of NANOGP8 or NANOG enhances the cytotoxicity of BH3 mimetics that target BCL-2 family members. Gene therapy targeting the NANOGs may increase the efficacy of BH3 mimetics in colorectal carcinoma.