Salubrinal在ABT737诱导的人肝癌细胞Sk-Hep1凋亡中的作用及机制
|
摘要
目的探索ABT737诱导的Sk-Hep1人肝癌细胞凋亡中salubrinal的作用其机制。方法联合应用ABT737与salubrinal后,转染eIF2αS51A质粒(51位丝氨酸不能磷酸化突变体)、eIF2αS51D(51位丝氨酸磷酸化突变体)质粒进入Sk-Hep1人肝癌细胞中,Western Blot检测PARP剪切蛋白、DNA损伤情况、p-eIF2α、eIF2α蛋白表达。结果在Sk-Hep1人肝癌细胞中联合应用salubrinal与ABT737后,与对照组相比,剪切的PARP蛋白表达量明显下调,磷酸化的H2AX蛋白表达量明显下调(P <0.05);p-eIF2α、eIF2α蛋白表达水平没有明显变化。转染eIF2αS51A、eIF2αS51D质粒后,与对照组相比,剪切的PARP、p-eIF2α蛋白表达水平无明显变化(P> 0.05)。结论 Salubrinal对ABT737诱导的细胞凋亡和损伤具有拮抗作用,该拮抗作用可能与eIF2α通路无关。
Objective To investigate the effects and its mechanism of salubrinal on ABT737-Hep1. Methods After the combined application of ABT737 and salubrinal,the plasmids of eIF2αS51 A(51 serine non-phosphorylated mutant)and eIF2αS51 D(51 Hep1 human hepatocellular carcinoma cells. Then,the cleavaged PARP protein,DNA damage,the p-eIF2α proteins and eIF2α proteins were measured by Western Blot-analyses. Results After the combined application of salubrinal and ABT737 in Sk-Hep1 human hepatocellular carcinoma cells,compared with the control group,the expressions of PARP cleavage proteins and phosphorylated H2 AX proteins were both significantly down-regulated(P < 0.05). There were no significant changes in the expres-sion levels of p-eIF2α and eIF2α proteins. After transfection of plasmids of eIF2αS51 A and eIF2αS51 D,there were no significant changes in the expression levels of cleavaged PARP and p-eIF2α protein compared with the control group(P > 0.05). Conclusion Salubrinal has an antagonistic action on apoptosis and DNA injury induced by ABT737,which may be unrelated to the eIF2α pathway.
Objective To investigate the effects and its mechanism of salubrinal on ABT737-Hep1. Methods After the combined application of ABT737 and salubrinal,the plasmids of eIF2αS51 A(51 serine non-phosphorylated mutant)and eIF2αS51 D(51 Hep1 human hepatocellular carcinoma cells. Then,the cleavaged PARP protein,DNA damage,the p-eIF2α proteins and eIF2α proteins were measured by Western Blot-analyses. Results After the combined application of salubrinal and ABT737 in Sk-Hep1 human hepatocellular carcinoma cells,compared with the control group,the expressions of PARP cleavage proteins and phosphorylated H2 AX proteins were both significantly down-regulated(P < 0.05). There were no significant changes in the expres-sion levels of p-eIF2α and eIF2α proteins. After transfection of plasmids of eIF2αS51 A and eIF2αS51 D,there were no significant changes in the expression levels of cleavaged PARP and p-eIF2α protein compared with the control group(P > 0.05). Conclusion Salubrinal has an antagonistic action on apoptosis and DNA injury induced by ABT737,which may be unrelated to the eIF2α pathway.
引文
[1]朱芳方,陈海清,赖己创,等.超声微泡介导质粒转染对人肝癌Hep G2细胞迁移、侵袭及克隆能力的影响[J].实用医学杂志,2017,33(16):2629-2633.
[2]秦蜀,陈润,赵晓芳,等.ATF4在CCl4和LPS/D-Gal N介导小鼠肝损伤中的保护作用[J].实用医学杂志,2018,32(13):2146-2150.
[3]PETROS A M,DINGES J,AUGERI D J,et al.Discovery of a potent inhibitor of the antiapoptotic protein Bcl-x L from NMRand parallel synthesis[J].J Med Chem,2006,49(2):656-663.
[4]XU Y,GAO W,ZHANG Y,et al.ABT737 reverses cisplatin resistance by targeting glucose metabolism of humanovarian cancer cells[J].Int J Oncol,2018,53(3):1055-1068.
[5]YU Y,XU L,QI L,et al.ABT737 induces mitochondrial pathway apoptosis and mitophagy by regulating DRP1-dependent mitochondrial fission in human ovarian cancer cells[J].Biomed Pharmacother,2017,96:22-29.
[6]KARPEL-MASSLER G,ISHIDA C T,ZHANG Y,et al.Targeting intrinsic apoptosis and other forms of cell death by BH3-mimetics in glioblastoma[J].Expert Opin Drug Discov,2017,12(10):1031-1040.
[7]石少敏,王静,于杜娟,等.Bcl-2抑制剂ABT737增加GSK3抑制剂SB216763引起的A549细胞凋亡[J].中国实验诊断学,2016,20(10):1619-1623.
[8]王焕,张丽娟,刘穗玲,等.ABT737通过激活JNK/c-Jun通路上调Bim诱导宫颈癌细胞凋亡[J].中山大学学报(医学科学版),2013,34(5):695-702.
[9]DO V H,WALTON S,CATT S,et al.Does the addition of salubrinal to in vitro maturation medium enhance bovine blastocyst yields and embryo cryotolerance?[J].Cryo Letters,2018,39(3):219-226.
[10]BOYCE M,BRYANT K F,JOUSSE C,et al.A selective inhibitor of e IF2 apha dephosphorylation protects cells from ER stress[J].Science,2005,307(5711):935-939.
[11]陈永华,杨文明,江海林,等.Salubrinal对铜负荷诱导的PERK/e IF2α内质网应激信号通路介导肝细胞凋亡的干预作用[J].中国生物化学与分子生物学报,2018,34(9):990-996.
[12]刘利娟,陈瑶,周德生,等.Salubrinal对大鼠脑缺血再灌注模型LC3-Ⅱm RNA及LC3-Ⅱ/LC3-Ⅰm RNA的影响[J].广东医学,2017,38(4):509-513.
[13]YU W,XIANG Y,LUO G,et al.Salubrinal enhances doxorubicin sensitivity in humancholangiocarcinoma cells through promoting DNA damage[J].Cancer Biother Radiopharm,2018,33(6):258-265.
[14]ANUNCIBAY-SOTO B,PéREZ-RODRIGUEZ D,SANTOS-GALDIANO M,et al.Salubrinal and robenacoxib treatment after global cerebral ischemia.Exploringthe interactions between ER stress and inflammation[J].Biochem Pharmacol,2018,151:26-37.
[15]GONZáLEZ-ALMELA E,WILLIAMS H,SANZ M A,et al.The initiation factorse IF2,e IF2A,e IF2D,e IF4A,and e IF4Gare not involved in translation driven by hepatitis C virus IRESin human cells[J].Front Microbiol,2018,9:207.
[16]HU Y,LU X,XU Y,et al.Salubrinal attenuated retinal neovascularization by inhibiting CHOP-HIF1α-VEGF pathways[J].Oncotarget,2017,8(44):77219-77232.
[17]RANI S,SREENIVASAIAH P K,CHO C,et al.Salubrinal alleviates pressure overload-induced cardiac hypertrophy by inhibiting endoplasmic reticulum stress pathway[J].Mol Cells,2017,40(1):66-72.
[18]ANUNCIBAY-SOTO B,SANTOS-GALDIANO M,FERNáNDEZ-L?PEZ A.Neuroprotection by salubrinal treatment in global cerebral ischemia[J].Neural Regen Res,2016,11(11):1744-1745.
[19]CHEN S,SUN C,GU H,et al.Salubrinal protects against clostridium difficile toxin B-induced CT26 cell death[J].Acta Biochim Biophys Sin(Shanghai),2017,49(3):228-237.
[20]李云,郑广瑛,刘玥,等.Salubrinal对人晶状体上皮细胞中内质网应激的影响[J].中华眼科杂志,2016,52(6):437-443.
[21]BASTOLA P,NEUMS L,SCHOENEN F J,et al.VCP inhibitors induce endoplasmicreticulum stress,cause cell cycle arrest,trigger caspase-mediated cell death and synergistically kill ovarian cancer cells in combination with salubrinal[J].Mol Oncol,2016,10(10):1559-1574.
[22]JEON Y J,KIM J H,SHIN J I,et al.Salubrinal-mediated upregulation of e IF2alpha phosphorylation increases doxorubicin sensitivity in MCF-7/ADR cells[J].Mol Cells,2016,39(2):129-135.
[23]ZHAO X,ZHANG C,ZHOU H,et al.Synergistic antitumor activity of the combination of salubrinal and rapamycin against human cholangiocarcinoma cells[J].Oncotarget,2016,7(51):85492-85501.
[24]XIE Q,SU J,JIAO B,et al.ABT737 reverses cisplatin resistance by regulating ER-mitochondria Ca2+signal transduction in human ovarian cancer cells[J].Int J Oncol,2016,49(6):2507-2519.
[2]秦蜀,陈润,赵晓芳,等.ATF4在CCl4和LPS/D-Gal N介导小鼠肝损伤中的保护作用[J].实用医学杂志,2018,32(13):2146-2150.
[3]PETROS A M,DINGES J,AUGERI D J,et al.Discovery of a potent inhibitor of the antiapoptotic protein Bcl-x L from NMRand parallel synthesis[J].J Med Chem,2006,49(2):656-663.
[4]XU Y,GAO W,ZHANG Y,et al.ABT737 reverses cisplatin resistance by targeting glucose metabolism of humanovarian cancer cells[J].Int J Oncol,2018,53(3):1055-1068.
[5]YU Y,XU L,QI L,et al.ABT737 induces mitochondrial pathway apoptosis and mitophagy by regulating DRP1-dependent mitochondrial fission in human ovarian cancer cells[J].Biomed Pharmacother,2017,96:22-29.
[6]KARPEL-MASSLER G,ISHIDA C T,ZHANG Y,et al.Targeting intrinsic apoptosis and other forms of cell death by BH3-mimetics in glioblastoma[J].Expert Opin Drug Discov,2017,12(10):1031-1040.
[7]石少敏,王静,于杜娟,等.Bcl-2抑制剂ABT737增加GSK3抑制剂SB216763引起的A549细胞凋亡[J].中国实验诊断学,2016,20(10):1619-1623.
[8]王焕,张丽娟,刘穗玲,等.ABT737通过激活JNK/c-Jun通路上调Bim诱导宫颈癌细胞凋亡[J].中山大学学报(医学科学版),2013,34(5):695-702.
[9]DO V H,WALTON S,CATT S,et al.Does the addition of salubrinal to in vitro maturation medium enhance bovine blastocyst yields and embryo cryotolerance?[J].Cryo Letters,2018,39(3):219-226.
[10]BOYCE M,BRYANT K F,JOUSSE C,et al.A selective inhibitor of e IF2 apha dephosphorylation protects cells from ER stress[J].Science,2005,307(5711):935-939.
[11]陈永华,杨文明,江海林,等.Salubrinal对铜负荷诱导的PERK/e IF2α内质网应激信号通路介导肝细胞凋亡的干预作用[J].中国生物化学与分子生物学报,2018,34(9):990-996.
[12]刘利娟,陈瑶,周德生,等.Salubrinal对大鼠脑缺血再灌注模型LC3-Ⅱm RNA及LC3-Ⅱ/LC3-Ⅰm RNA的影响[J].广东医学,2017,38(4):509-513.
[13]YU W,XIANG Y,LUO G,et al.Salubrinal enhances doxorubicin sensitivity in humancholangiocarcinoma cells through promoting DNA damage[J].Cancer Biother Radiopharm,2018,33(6):258-265.
[14]ANUNCIBAY-SOTO B,PéREZ-RODRIGUEZ D,SANTOS-GALDIANO M,et al.Salubrinal and robenacoxib treatment after global cerebral ischemia.Exploringthe interactions between ER stress and inflammation[J].Biochem Pharmacol,2018,151:26-37.
[15]GONZáLEZ-ALMELA E,WILLIAMS H,SANZ M A,et al.The initiation factorse IF2,e IF2A,e IF2D,e IF4A,and e IF4Gare not involved in translation driven by hepatitis C virus IRESin human cells[J].Front Microbiol,2018,9:207.
[16]HU Y,LU X,XU Y,et al.Salubrinal attenuated retinal neovascularization by inhibiting CHOP-HIF1α-VEGF pathways[J].Oncotarget,2017,8(44):77219-77232.
[17]RANI S,SREENIVASAIAH P K,CHO C,et al.Salubrinal alleviates pressure overload-induced cardiac hypertrophy by inhibiting endoplasmic reticulum stress pathway[J].Mol Cells,2017,40(1):66-72.
[18]ANUNCIBAY-SOTO B,SANTOS-GALDIANO M,FERNáNDEZ-L?PEZ A.Neuroprotection by salubrinal treatment in global cerebral ischemia[J].Neural Regen Res,2016,11(11):1744-1745.
[19]CHEN S,SUN C,GU H,et al.Salubrinal protects against clostridium difficile toxin B-induced CT26 cell death[J].Acta Biochim Biophys Sin(Shanghai),2017,49(3):228-237.
[20]李云,郑广瑛,刘玥,等.Salubrinal对人晶状体上皮细胞中内质网应激的影响[J].中华眼科杂志,2016,52(6):437-443.
[21]BASTOLA P,NEUMS L,SCHOENEN F J,et al.VCP inhibitors induce endoplasmicreticulum stress,cause cell cycle arrest,trigger caspase-mediated cell death and synergistically kill ovarian cancer cells in combination with salubrinal[J].Mol Oncol,2016,10(10):1559-1574.
[22]JEON Y J,KIM J H,SHIN J I,et al.Salubrinal-mediated upregulation of e IF2alpha phosphorylation increases doxorubicin sensitivity in MCF-7/ADR cells[J].Mol Cells,2016,39(2):129-135.
[23]ZHAO X,ZHANG C,ZHOU H,et al.Synergistic antitumor activity of the combination of salubrinal and rapamycin against human cholangiocarcinoma cells[J].Oncotarget,2016,7(51):85492-85501.
[24]XIE Q,SU J,JIAO B,et al.ABT737 reverses cisplatin resistance by regulating ER-mitochondria Ca2+signal transduction in human ovarian cancer cells[J].Int J Oncol,2016,49(6):2507-2519.