内质网应激IRE-1/XBP-1参与调控肝癌进展的机制研究
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摘要
目的探讨内质网应激(ERS)调控内质网核信号转导蛋白a1(Endoplasmic reticulum to nucleus signaling 1,IRE-1) IRE-1/X-盒-结合蛋白-1(X box Binding Protein 1,XBP-1) XBP-1参与肝癌进展的机制研究。方法用无水乙醇诱导正常肝细胞LO2、高分化肝癌细胞株Hep G2及高转移能力肝癌细胞株SMMC-7721细胞48 h获得内质网应激细胞模型;采用CCK-8法检测细胞的增殖情况;采用Hoechst 33258染色检测细胞凋亡情况;采用Real-time PCR和Western blot方法检测无水乙醇处理前后IRE-1、XBP-1、DNA损伤诱导基因153(CHOP)、Cleaved-caspase 12和Bcl-2 mRNA和蛋白含量表达。结果无水乙醇诱导LO2、Hep G2、和SMMC-7721细胞发生内质网应激后,细胞存活率显著降低,细胞凋亡明显升高,差异有统计学意义(P <0. 05)。无水乙醇诱导LO2、Hep G2、和SMMC-7721细胞发生内质网应激后,LO2、Hep G2、和SMMC-7721细胞内IRE-1、XBP-1、CHOP、Cleaved-caspase 12 mRNA和蛋白表达含量显著升高,其中细胞内mRNA和蛋白含量表达的高低次序为SMMC-7721、Hep G2和LO2细胞,差异有统计学意义(P <0. 05); Bcl-2 mRNA和蛋白表达含量显著降低,细胞内mRNA和蛋白含量表达的高低次序为LO2、Hep G2和SMMC-7721细胞,差异有统计学意义(P <0. 05)。结论内质网应激发生后LO2、Hep G2和SMMC-7721细胞中IRE-1/XBP-1的表达水平显著升高,有效参与调控肝癌细胞的增殖发展。
Objective To investigate the regulation of endoplasmic reticulum stress( ERS) in endoplasmic reticulum to nucleus signaling( IRE-1) IRE-1/X-box-binding protein-1( X) Box Binding Protein 1,XBP-1) XBP-1 is involved in the mechanism of liver cancer progression. Methods The endoplasmic reticulum stress cell model was obtained by inducing normal liver cell LO2,well-differentiated hepatocellular carcinoma cell line Hep G2 and high metastatic liver cancer cell line SMMC-7721 cells with absolute ethanol for 48 hours. Cell proliferation was detected by CCK-8 method. Apoptosis was detected by Hoechst 33258 staining; Real-time PCR and Western blot were used to detect IRE-1,XBP-1,DNA damage-inducing gene 153( CHOP),Cleavedcaspase 12 and Bcl-2 before and after absolute ethanol treatment. mRNA and protein content expression. Results After ethanolinduced LO2,Hep G2,and SMMC-7721 cells showed endoplasmic reticulum stress,cell survival rate was significantly decreased,and apoptosis was significantly increased( P < 0. 05). Expression of IRE-1,XBP-1,CHOP and Cleaved-caspase 12 mRNA and protein in LO2,Hep G2,and SMMC-7721 cells induced by endoplasmic reticulum stress in LO2,Hep G2,and SMMC-7721 cells induced by absolute ethanol Significantly increased,the order of mRNA and protein expression in cells was SMMC-7721,Hep G2 and LO2 cells,the difference was statistically significant( P < 0. 05); Bcl-2 mRNA and protein expression levels were significantly decreased,intracellular mRNA The order of protein and protein expression was LO2,Hep G2 and SMMC-7721 cells,and the difference was statistically significant( P < 0. 05). Conclusion The expression levels of IRE-1/XBP-1 in LO2,Hep G2 and SMMC-7721 cells are significantly increased after endoplasmic reticulum stress,which is effective in regulating the proliferation of hepatoma cells.
Objective To investigate the regulation of endoplasmic reticulum stress( ERS) in endoplasmic reticulum to nucleus signaling( IRE-1) IRE-1/X-box-binding protein-1( X) Box Binding Protein 1,XBP-1) XBP-1 is involved in the mechanism of liver cancer progression. Methods The endoplasmic reticulum stress cell model was obtained by inducing normal liver cell LO2,well-differentiated hepatocellular carcinoma cell line Hep G2 and high metastatic liver cancer cell line SMMC-7721 cells with absolute ethanol for 48 hours. Cell proliferation was detected by CCK-8 method. Apoptosis was detected by Hoechst 33258 staining; Real-time PCR and Western blot were used to detect IRE-1,XBP-1,DNA damage-inducing gene 153( CHOP),Cleavedcaspase 12 and Bcl-2 before and after absolute ethanol treatment. mRNA and protein content expression. Results After ethanolinduced LO2,Hep G2,and SMMC-7721 cells showed endoplasmic reticulum stress,cell survival rate was significantly decreased,and apoptosis was significantly increased( P < 0. 05). Expression of IRE-1,XBP-1,CHOP and Cleaved-caspase 12 mRNA and protein in LO2,Hep G2,and SMMC-7721 cells induced by endoplasmic reticulum stress in LO2,Hep G2,and SMMC-7721 cells induced by absolute ethanol Significantly increased,the order of mRNA and protein expression in cells was SMMC-7721,Hep G2 and LO2 cells,the difference was statistically significant( P < 0. 05); Bcl-2 mRNA and protein expression levels were significantly decreased,intracellular mRNA The order of protein and protein expression was LO2,Hep G2 and SMMC-7721 cells,and the difference was statistically significant( P < 0. 05). Conclusion The expression levels of IRE-1/XBP-1 in LO2,Hep G2 and SMMC-7721 cells are significantly increased after endoplasmic reticulum stress,which is effective in regulating the proliferation of hepatoma cells.
引文
[1]陶建平,陈忠.Sirt1抑制内质网依赖的巨噬细胞凋亡[J].现代医学,2017,45(8):1059-1064.
[2]于梦尧,应长江,周冬梅,等.阿利沙坦酯对糖尿病大鼠心肌内质网应激和SERCA2a表达的影响[J].现代医学,2017,45(12):1709-1713.
[3]李冰心,周小兵,刘梦捷,等.高糖对肾小管上皮细胞内质网应激的影响及冬虫夏草的干预作用[J].中国医院药学杂志,2016,36(22):1977-1980.
[4]谢慧臣,罗丽华,刘芬,等.头顶一颗珠水提取物对非酒精性脂肪肝大鼠肝脏结构及功能的影响[J].中国医院药学杂志,2017,37(3):248-253.
[5]李克跃,石承先,汤可立,等.川芎嗪对人肝癌细胞TGF-β1/Smads通路的影响[J].东南大学学报(医学版),2017,36(4):554-558.
[6]董凯楠,黄忻,邢文英.内质网应激对胃癌细胞迁移与侵袭的影响[J].世界华人消化杂志,2016,24(10):1485-1491.
[7]郭佳培,吴景华,李雷雷,等.AKT-GSK-3β信号途径在下调肝癌细胞化疗敏感性中的作用研究[J].标记免疫分析与临床,2016,23(3):316-318.
[8]张娟,彭晓蔓,刘朝奇.内质网应激在非酒精性脂肪性肝炎中的致病机制[J].中国免疫学杂志,2017,33(7):1112-1114.
[9]李军汉,李恩,张仲阳,等.运动对非酒精性脂肪肝形成中内质网应激PERK/e IF-2a、IRE-1/XBP-1通路的影响[J].首都体育学院学报,2016,28(5):459-462.
[10]李军汉,孙君志,李恩,等.运动和饮食调整对非酒精性脂肪肝大鼠内质网应激PERK/e IF-2a和IRE-1/XBP-1通路的影响[J].成都体育学院学报,2016,42(6):110-113,126.
[11]谷晓峰,邓学峰,马群风,等.RNF13通过IRE1/XBP-1通路调控内质网应激介导的细胞凋亡[J].现代生物医学进展,2016,16(5):801-805.
[12]李雷雷,郭彬,郭佳培,等.M2型肿瘤相关巨噬细胞通过p53途径下调肝癌化疗敏感性的机制研究[J].标记免疫分析与临床,2017,24(3):308-313.
[13]王利平,符鹏程,秦玲.蛋白酶体抑制剂通过内质网应激影响前列腺癌DU145细胞的生长[J].现代肿瘤医学,2016,24(18):2857-2862.
[14]李利波,潘娅,陈腾祥.内质网应激前后肝癌细胞中C/EBP同源蛋白及X-盒-结合蛋白-1的表达[J].贵州医科大学学报,2016,41(9):1033-1036.
[15]丁娟,纪璐璐,徐亚廷,等.高糖诱导滋养层细胞内质网应激及凋亡[J].中国组织化学与细胞化学杂志,2016,25(1):13-17.
[16]张玮,申文豪,李阳,等.三七总皂苷联合顺铂对肝癌细胞Hep G2和SMMC-7721增殖、迁移、侵袭及凋亡的影响[J].东南大学学报(医学版),2017,36(3):383-389.
[2]于梦尧,应长江,周冬梅,等.阿利沙坦酯对糖尿病大鼠心肌内质网应激和SERCA2a表达的影响[J].现代医学,2017,45(12):1709-1713.
[3]李冰心,周小兵,刘梦捷,等.高糖对肾小管上皮细胞内质网应激的影响及冬虫夏草的干预作用[J].中国医院药学杂志,2016,36(22):1977-1980.
[4]谢慧臣,罗丽华,刘芬,等.头顶一颗珠水提取物对非酒精性脂肪肝大鼠肝脏结构及功能的影响[J].中国医院药学杂志,2017,37(3):248-253.
[5]李克跃,石承先,汤可立,等.川芎嗪对人肝癌细胞TGF-β1/Smads通路的影响[J].东南大学学报(医学版),2017,36(4):554-558.
[6]董凯楠,黄忻,邢文英.内质网应激对胃癌细胞迁移与侵袭的影响[J].世界华人消化杂志,2016,24(10):1485-1491.
[7]郭佳培,吴景华,李雷雷,等.AKT-GSK-3β信号途径在下调肝癌细胞化疗敏感性中的作用研究[J].标记免疫分析与临床,2016,23(3):316-318.
[8]张娟,彭晓蔓,刘朝奇.内质网应激在非酒精性脂肪性肝炎中的致病机制[J].中国免疫学杂志,2017,33(7):1112-1114.
[9]李军汉,李恩,张仲阳,等.运动对非酒精性脂肪肝形成中内质网应激PERK/e IF-2a、IRE-1/XBP-1通路的影响[J].首都体育学院学报,2016,28(5):459-462.
[10]李军汉,孙君志,李恩,等.运动和饮食调整对非酒精性脂肪肝大鼠内质网应激PERK/e IF-2a和IRE-1/XBP-1通路的影响[J].成都体育学院学报,2016,42(6):110-113,126.
[11]谷晓峰,邓学峰,马群风,等.RNF13通过IRE1/XBP-1通路调控内质网应激介导的细胞凋亡[J].现代生物医学进展,2016,16(5):801-805.
[12]李雷雷,郭彬,郭佳培,等.M2型肿瘤相关巨噬细胞通过p53途径下调肝癌化疗敏感性的机制研究[J].标记免疫分析与临床,2017,24(3):308-313.
[13]王利平,符鹏程,秦玲.蛋白酶体抑制剂通过内质网应激影响前列腺癌DU145细胞的生长[J].现代肿瘤医学,2016,24(18):2857-2862.
[14]李利波,潘娅,陈腾祥.内质网应激前后肝癌细胞中C/EBP同源蛋白及X-盒-结合蛋白-1的表达[J].贵州医科大学学报,2016,41(9):1033-1036.
[15]丁娟,纪璐璐,徐亚廷,等.高糖诱导滋养层细胞内质网应激及凋亡[J].中国组织化学与细胞化学杂志,2016,25(1):13-17.
[16]张玮,申文豪,李阳,等.三七总皂苷联合顺铂对肝癌细胞Hep G2和SMMC-7721增殖、迁移、侵袭及凋亡的影响[J].东南大学学报(医学版),2017,36(3):383-389.