热休克蛋白90α通过增强钙库操作性钙离子内流活性促进肝癌细胞转移
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摘要
目的探讨细胞外热休克蛋白90α(eHsp90α)对肝癌细胞转移的影响及机制。方法 Westernblot检测细胞培养基上清中eHsp90α的水平和细胞中基质相互作用分子1(STIM1)蛋白表达,Confocal检测钙库操作性钙离子内流(SOCE)活性,免疫荧光双染检测STIM1和ORAI1的相互作用水平,Transwell和划痕实验检测细胞转移能力。结果肝癌细胞系中e Hsp90α较正常肝细胞系LO-2显著增加;高度转移能力的MHCC87H和HCCLM3细胞中e Hsp90α水平、STIM1蛋白表达和SOCE活性较低转移能力的HepG2、HL-7702和SNU-449显著升高。hrHsp90α可以显著增加HepG2的转移能力、STIM1蛋白变化和SOCE活性。敲低STIM1蛋白表达或抑制SOCE活性,可以抑制MHCC87H的转移能力和hrHsp90α诱导的HepG2转移能力增加。结论eHsp90α可促进肝癌细胞的转移,其机制是通过诱导STIM1蛋白表达,进而增加SOCE活性。
Objective To investigate the effect and mechanism of extracellular heat shock protein 90 a(eHsp90α) on the metastasis of hepatocellular carcinoma cells. Method The level of eHsp90α in the supernatant of cell culture medium and stromal interaction molecule 1(STIM1) protein in the cells were detected by western blot assay. The confocal assay was used to detect SOCE activity. Immunofluorescence double staining was used to detect the interaction between STIM1 and ORAI1. The transwell and wound-healing assay was performed to detect the metastasis of hepatocellular carcinoma cells. Results The expression of eHsp90α in hepatocellular carcinoma cell lines was significantly higher than that in normal liver cell line LO-2. The expression of eHsp90α and STIM1, and SOCE activity were significantly increased in MHC87 H and HCCLM3 cells, with high metastatic ability than that in HepG2, HL-7702 and SNU-449 cells. hrHsp90 a could significantly increase HepG2 metastasis, as well as STIM1 expression and SOCE activity. The metastasis of MHC87 H cell and eHsp90 a-induced HepG2 cell was inhibited when STIM1 was knockdown or SOCE activity was suppressed. Conclusion eHsp90α can promote the metastasis of hepatocarcinoma cells by inducing STIM1 expression to increase SOCE activity.
Objective To investigate the effect and mechanism of extracellular heat shock protein 90 a(eHsp90α) on the metastasis of hepatocellular carcinoma cells. Method The level of eHsp90α in the supernatant of cell culture medium and stromal interaction molecule 1(STIM1) protein in the cells were detected by western blot assay. The confocal assay was used to detect SOCE activity. Immunofluorescence double staining was used to detect the interaction between STIM1 and ORAI1. The transwell and wound-healing assay was performed to detect the metastasis of hepatocellular carcinoma cells. Results The expression of eHsp90α in hepatocellular carcinoma cell lines was significantly higher than that in normal liver cell line LO-2. The expression of eHsp90α and STIM1, and SOCE activity were significantly increased in MHC87 H and HCCLM3 cells, with high metastatic ability than that in HepG2, HL-7702 and SNU-449 cells. hrHsp90 a could significantly increase HepG2 metastasis, as well as STIM1 expression and SOCE activity. The metastasis of MHC87 H cell and eHsp90 a-induced HepG2 cell was inhibited when STIM1 was knockdown or SOCE activity was suppressed. Conclusion eHsp90α can promote the metastasis of hepatocarcinoma cells by inducing STIM1 expression to increase SOCE activity.
引文
[1]ASRANI S K,DEVARBHAVI H,EATON J,et al.Burden of liver diseases in the world[J].J Hepatol,2018.pii:SO168-8278(18)32388-2.
[2]闵家祺,邬林泉.干扰素诱导跨膜蛋白3诱导上皮细胞间质转化促进肝癌侵袭转移[J].实用医学杂志,2018,34(15):2485-2490.
[3]MAO X,WONG S Y,TSE E Y,et al.Mechanisms through which hypoxia-induced caveolin-1 drives tumorigenesis and metastasis in hepatocellular carcinoma[J].Cancer Res,2016,76(24):7242-7253.
[4]TANG B,QI G,TANG F,et al.Aberrant JMJD3 expression upregulates slug to promote migration,invasion,and stem celllike behaviors in hepatocellular carcinoma[J].Cancer Res,2016,76(22):6520-6532.
[5]YANG N,TANG Y,WANG F,et al.Blockade of store-operated Ca(2+)entry inhibits hepatocarcinoma cell migration and invasion by regulating focal adhesion turnover[J].Cancer Lett,2013,330(2):163-169.
[6]叶红丽,肖凤.基质交感分子1在肿瘤细胞生长中的研究进展[J].实用临床医学,2013(8):123-125,128.
[7]VARTHOLOMAIOU E,MADON-SIMON M,HAGMANN S,et al.Cytosolic Hsp90alpha and its mitochondrial isoform Trap1are differentially required in a breast cancer model[J].Oncotarget,2017,8(11):17428-17442.
[8]CHATTERJEE S,BURNS T F.Targeting heat shock proteins in cancer:a promising therapeutic approach[J].Int J Mol Sci,2017,18(9).pii:E1978.
[9]DONG H,ZOU M,BHATIA A,et al.Breast cancer MDA-MB-231 cells use secreted heat shock protein-90alpha(hsp90alpha)to survive a hostile hypoxic environment[J].Sci Rep,2016,6:20605.
[10]BOHONOWYCH J E,HANCE M W,NOLAN K D,et al.Extracellular Hsp90 mediates an NF-kappaB dependent inflammatory stromal program:implications for the prostate tumor microenvironment[J].Prostate,2014,74(4):395-407.
[11]SOURBIER C.Plasma HSP90alpha and liver cancer:a potential biomarker?[J].EBioMedicine,2017,25:7-8.
[12]KOU X,JIANG X,LIU H,et al.Simvastatin functions as a heat shock protein 90 inhibitor against triple-negative breast cancer[J].Cancer Sci,2018,109(10):3272-3284.
[13]THAKUR M K,HEILBRUN L K,SHENG S,et al.A phase IItrial of ganetespib,a heat shock protein 90 Hsp90)inhibitor,in patients with docetaxel-pretreated metastatic castrate-resistant prostate cancer(CRPC)-a prostate cancer clinical trials consortium(PCCTC)study[J].Invest New Drugs,2016,34(1):112-118.
[14]PEDERSEN K S,KIM G P,FOSTER N R,et al.Phase II trial of gemcitabine and tanespimycin(17AAG)in metastatic pancreatic cancer:a Mayo Clinic Phase II Consortium study[J].Invest New Drugs,2015,33(4):963-968.
[15]YU C,TANG W,WANG Y,et al.Downregulation of ACE2/Ang-(1-7)/Mas axis promotes breast cancer metastasis by enhancing store-operated calcium entry[J].Cancer Lett,2016,376(2):268-277.
[16]BABAER D,AMARA S,IVY M,et al.High salt induces P-glycoprotein mediated treatment resistance in breast cancer cells through store operated calcium influx[J].Oncotarget,2018,9(38):25193-25205.
[17]WANG Y,HE J,JIANG H,et al.Nicotine enhances storeoperated calcium entry by upregulating HIF1alpha and SOCC components in nonsmall cell lung cancer cells[J].Oncol Rep,2018,40(4):2097-2104.
[18]SUN X,WEI Q,CHENG J,et al.Enhanced Stim1 expression is associated with acquired chemo-resistance of cisplatin in osteosarcoma cells[J].Hum Cell,2017,30(3):216-225.
[19]ALI E S,RYCHKOV G Y,BARRITT G J.Metabolic disorders and cancer:hepatocyte store-operated ca(2+)channels in nonalcoholic fatty liver disease[J].Adv Exp Med Biol,2017,993:595-621.
[20]PRAKRIYA M,LEWIS R S.Store-operated calcium channels[J].Physiol Rev,2015,95(4):1383-1436.
[21]SAINT F S,MAUS M,VAETH M,et al.STIM1 and STIM2mediate cancer-induced inflammation in t cell acute lymphoblastic leukemia[J].Cell Rep,2018,24(11):3045-3060.
[22]吴冰,林天骥,阮仕娟,等.STIM1对乳腺癌细胞移动的作用及机制[J].实用医学杂志,2017,33(9):1373-1376.
[2]闵家祺,邬林泉.干扰素诱导跨膜蛋白3诱导上皮细胞间质转化促进肝癌侵袭转移[J].实用医学杂志,2018,34(15):2485-2490.
[3]MAO X,WONG S Y,TSE E Y,et al.Mechanisms through which hypoxia-induced caveolin-1 drives tumorigenesis and metastasis in hepatocellular carcinoma[J].Cancer Res,2016,76(24):7242-7253.
[4]TANG B,QI G,TANG F,et al.Aberrant JMJD3 expression upregulates slug to promote migration,invasion,and stem celllike behaviors in hepatocellular carcinoma[J].Cancer Res,2016,76(22):6520-6532.
[5]YANG N,TANG Y,WANG F,et al.Blockade of store-operated Ca(2+)entry inhibits hepatocarcinoma cell migration and invasion by regulating focal adhesion turnover[J].Cancer Lett,2013,330(2):163-169.
[6]叶红丽,肖凤.基质交感分子1在肿瘤细胞生长中的研究进展[J].实用临床医学,2013(8):123-125,128.
[7]VARTHOLOMAIOU E,MADON-SIMON M,HAGMANN S,et al.Cytosolic Hsp90alpha and its mitochondrial isoform Trap1are differentially required in a breast cancer model[J].Oncotarget,2017,8(11):17428-17442.
[8]CHATTERJEE S,BURNS T F.Targeting heat shock proteins in cancer:a promising therapeutic approach[J].Int J Mol Sci,2017,18(9).pii:E1978.
[9]DONG H,ZOU M,BHATIA A,et al.Breast cancer MDA-MB-231 cells use secreted heat shock protein-90alpha(hsp90alpha)to survive a hostile hypoxic environment[J].Sci Rep,2016,6:20605.
[10]BOHONOWYCH J E,HANCE M W,NOLAN K D,et al.Extracellular Hsp90 mediates an NF-kappaB dependent inflammatory stromal program:implications for the prostate tumor microenvironment[J].Prostate,2014,74(4):395-407.
[11]SOURBIER C.Plasma HSP90alpha and liver cancer:a potential biomarker?[J].EBioMedicine,2017,25:7-8.
[12]KOU X,JIANG X,LIU H,et al.Simvastatin functions as a heat shock protein 90 inhibitor against triple-negative breast cancer[J].Cancer Sci,2018,109(10):3272-3284.
[13]THAKUR M K,HEILBRUN L K,SHENG S,et al.A phase IItrial of ganetespib,a heat shock protein 90 Hsp90)inhibitor,in patients with docetaxel-pretreated metastatic castrate-resistant prostate cancer(CRPC)-a prostate cancer clinical trials consortium(PCCTC)study[J].Invest New Drugs,2016,34(1):112-118.
[14]PEDERSEN K S,KIM G P,FOSTER N R,et al.Phase II trial of gemcitabine and tanespimycin(17AAG)in metastatic pancreatic cancer:a Mayo Clinic Phase II Consortium study[J].Invest New Drugs,2015,33(4):963-968.
[15]YU C,TANG W,WANG Y,et al.Downregulation of ACE2/Ang-(1-7)/Mas axis promotes breast cancer metastasis by enhancing store-operated calcium entry[J].Cancer Lett,2016,376(2):268-277.
[16]BABAER D,AMARA S,IVY M,et al.High salt induces P-glycoprotein mediated treatment resistance in breast cancer cells through store operated calcium influx[J].Oncotarget,2018,9(38):25193-25205.
[17]WANG Y,HE J,JIANG H,et al.Nicotine enhances storeoperated calcium entry by upregulating HIF1alpha and SOCC components in nonsmall cell lung cancer cells[J].Oncol Rep,2018,40(4):2097-2104.
[18]SUN X,WEI Q,CHENG J,et al.Enhanced Stim1 expression is associated with acquired chemo-resistance of cisplatin in osteosarcoma cells[J].Hum Cell,2017,30(3):216-225.
[19]ALI E S,RYCHKOV G Y,BARRITT G J.Metabolic disorders and cancer:hepatocyte store-operated ca(2+)channels in nonalcoholic fatty liver disease[J].Adv Exp Med Biol,2017,993:595-621.
[20]PRAKRIYA M,LEWIS R S.Store-operated calcium channels[J].Physiol Rev,2015,95(4):1383-1436.
[21]SAINT F S,MAUS M,VAETH M,et al.STIM1 and STIM2mediate cancer-induced inflammation in t cell acute lymphoblastic leukemia[J].Cell Rep,2018,24(11):3045-3060.
[22]吴冰,林天骥,阮仕娟,等.STIM1对乳腺癌细胞移动的作用及机制[J].实用医学杂志,2017,33(9):1373-1376.