您的位置：山东大学 -> 科技期刊社 -> 《山东大学学报（医学版）》

山东大学学报（医学版） ›› 2015, Vol. 53 ›› Issue (6): 7-12.doi: 10.6040/j.issn.1671-7554.0.2014.811

• 基础医学 • 上一篇    下一篇

特异性下调Grp78表达增加肝癌细胞对厄洛替尼的敏感性

宋佳1, 李鑫2, 李丹2, 叶丽平1   

  1. 1. 辽宁医学院 病理生理学教研室, 辽宁 锦州 121001;
    2. 辽宁医学院 科学实验中心, 辽宁 锦州 121001
  • 收稿日期:2014-11-11 修回日期:2015-03-20 出版日期:2015-06-10 发布日期:2015-06-10
  • 通讯作者: 叶丽平。 E-mail:ye960519@126.com E-mail:ye960519@126.com
  • 基金资助:
    辽宁省博士启动基金(20101065)

Knockout Grp78 expression increases the sensitivity of hepatoma cells to erlotinib

SONG Jia1, LI Xin2, LI Dan2, YE Liping1   

  1. 1. Department of Pathophysiology, Liaoning Medical University, Jinzhou 121001, Liaoning, China;
    2. Science Laboratory Center, Liaoning Medical University, Jinzhou 121001, Liaoning, China
  • Received:2014-11-11 Revised:2015-03-20 Online:2015-06-10 Published:2015-06-10

PDF (PC)

244

摘要： 目的 探讨特异性下调Grp78表达后肝癌细胞对厄洛替尼敏感性的变化及其机制。方法 通过用siRNA技术特异性地下调肝癌细胞SMMC-7721中Grp78的表达并做Western blotting鉴定,MTT法检测不同浓度(0、2.5、5、10、20 μmol/L)厄洛替尼作用下肝癌细胞存活率的变化,流式细胞术检测细胞凋亡的变化,吖啶橙荧光染色法检测细胞凋亡的形态学变化,免疫印迹法检测AKT、p-AKT、ERK和p- ERK表达。结果 转染siRNA- Grp78后24、48、72 h均能明显抑制Grp78的表达,其中48 h抑制效果最明显(P<0.05)。与SMMC-7721/siControl相比,厄洛替尼对SMMC-7721/siRNA-Grp78细胞具有明显的增殖抑制作用和促凋亡作用,差异有统计学意义(P<0.05)。与SMMC-7721/siControl相比,厄洛替尼可明显降低SMMC-7721/siRNA-Grp78细胞中p- ERK和p-AKT的蛋白表达(P<0.05),但总ERK和AKT蛋白表达不受影响。结论 特异性下调肝癌细胞中Grp78的表达通过抑制ERK和AKT蛋白的磷酸化增强肝癌细胞对厄洛替尼的敏感性。

关键词: 肝癌, Grp78, 厄洛替尼

Abstract: Objective To explore the effects of knockout Grp78 expression on the sensitivity of SMMC-7721 cells to erlotinib and the molecular mechanism. Methods The Grp78 expression in SMMC-7721 cells was downregulated with siRNA technique and the expression was identified with Western blotting. The survival rates of cells treated with different concentrations of erlotinib (0, 2.5, 5, 10, 20 μmol/L) were assessed with MTT method. Cell apoptosis was detected with flow cytometry. The morphological changes of apoptosis were evaluated with acridine orange staining. The expression of AKT, p-AKT, ERK and p-ERK were determined with Western blotting. Results The siRNA transfection significantly inhibited Grp78 expression after 24, 48 and 72 hours, and the strongest inhibitive effect appeared after 48 hours (P<0.05). Compared with SMMC-7721/siControl cells, erlotinib significantly inhibited the proliferation and promoted apoptosis of SMMC-7721/siRNA-Grp78 cells (P<0.05), and markedly reduced the expression of p-ERK and p-AKT in SMMC-7721/siRNA-Grp78 cells (P<0.05), while the total protein expression of ERK and AKT were not affected. Conclusion Knockout Grp78 expression increases the sensitivity of hepatoma cells to erlotinib by decreasing the phosphorylation of ERK and AKT.

Key words: Erlotinib, Grp78, Hepatocellular carcinoma

中图分类号: 

  • R735
[1] Mittal S, El-Serag HB. Epidemiology of hepatocellular carcinoma: consider the population[J]. J Clin Gastroenterol, 2013, 47 Suppl:S2-S6.
[2] Hoshi S, Yamaguchi T, Kono C, et al. Recurrence of non-small cell lung cancer after successful treatment with gefitinib-report of three cases[J]. Gan To Kagaku Ryoho, 2004, 31(8):1209-1213.
[3] Pao W, Miller VA, Politi KA, et al. Acquired resistance of lung adenocarcinomas to gefitinib or erlotinib is associated with a second mutation in the EGFR kinase domain[J]. Plos Medicine, 2005, 29(3):57-62.
[4] Lee E, Nichols P, Spicer D, et al. GRP78 as a novel predictor of responsiveness to chemotherapy in breast cancer[J]. Cancer Res, 2006, 66(16):7849-7853.
[5] Reddy RK, Mao C, Baumeister P, et al. Endoplasmic reticulum chaperone protein GRP78 protects cells from apoptosis induced by topoisomerase inhibitors: role of ATP binding site in suppression of caspase-7 activation[J]. J Biol Chem, 2003, 278(23):20915-20924.
[6] Tsutsumi S, Namba T, Tanaka KI, et al. Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells[J]. Oncogene, 2006, 25(7):1018-1029.
[7] Metzger B, Chambeau L, Begon DY, et al. The human epidermal growth factor receptor (EGFR) gene in European patients with advanced colorectal cancer harbors infrequent mutations in its tyrosine kinase domain[J]. BMC Med Genet, 2011, 12: 144. doi: 10.1186/1471-2350-12-144.
[8] 徐立, 张昌卿, 冯凯涛, 等. 肝细胞癌及癌旁肝组织EGFR和EGFR vⅢ的表达及其临床意义[J]. 中山大学学报: 医学科学版, 2006, 27(4): 467- 471. XU Li, ZHANG Changqing, FENG Kaitao, et al. Clinical significance of EGFR and EGFRvⅢ expression in tumor and liver tissue of HCC[J]. Journal of Sun Yat-sen University: Medical Sciences, 2006, 27(4): 467- 471.
[9] Mueller KL, Powell K, Madden JM, et al. EGFR tyrosine 845 phosphorylation-dependent proliferation and transformation of breast cancer cells require activation of p38 MAPK[J]. Transl Oncol, 2012, 5(5): 327-334.
[10] Lindzen M, Carvalho S, Starr A, et al. A recombinant decoy comprising EGFR and ErbB-4 inhibits tumor growth and metastasis[J]. Oncogene, 2012, 31(30): 3505-3515.
[11] Donev IS, Wang W, Yamada T, et al. Transient PI3K inhibition induces apoptosis and overcomes HGF-mediated resistance to EGFR-TKIs in EGFR mutant lung cancer[J]. Clin Cancer Res, 2011, 17(8): 2260-2269.
[12] Hsu CH, Kang YK, Yang TS, et al. Bevacizumab with erlotinib as first-line therapy in Asian patients with advanced hepatocellular carcinoma: a multicenter phase II study[J]. Oncology, 2013, 85(1): 44-52.
[13] Fuchs BC, Hoshida Y, Fujii TE, et al. Epidermal growth factor receptor inhibition attenuates liver fibrosis and development of hepatocellular carcinoma[J]. Hepatology, 2014, 59(4): 1577-1590.
[14] Thomas MB, Chadha R, Glover K, et al. Phase 2 study of erlotinib in patients with unresectable hepatocellular carcinoma[J]. Cancer, 2007, 110(5): 1059-1067.
[15] Philip PA, Mahoney MR, Allmer C, et al. Phase II study of Erlotinib (OSI-774) in patients with advanced hepatocellular cancer[J]. J Clin Oncol, 2005, 23(27): 6657-6663.
[16] Su R, Li Z, Li H, et al. Grp78 promotes the invasion of hepatocellular carcinoma[J]. BMC Cancer, 2010, 10: 20. doi: 101186/1471-2407-10-20.
[17] Fu R, Yang P, Wu HL, et al. GRP78 secreted by colon cancer cells facilitates cell proliferation via PI3K/Akt signaling[J]. Asian Pac J Cancer Prev, 2014, 15(17): 7245-7249.
[18] Zhang LH, Yang XL, Zhang X, et al. Association of elevated GRP78 expression with increased astrocytoma malignancy via Akt and ERK pathways[J]. Brain Res, 2011, 1371: 23-31.
[1] 田丽君,桑玉洁,孙瑜婧,韩冰,秦成勇,祁建妮. 全身免疫炎症指数对原发性肝癌患者免疫检查点抑制剂治疗相关不良反应的预测价值[J]. 山东大学学报 (医学版), 2024, 62(6): 48-53.
[2] 靳新娟,左立平,邓展昊,李安宁,于德新. MRI影像组学对135例肝癌耐药蛋白PFKFB3的预测价值[J]. 山东大学学报 (医学版), 2023, 61(6): 79-86.
[3] 王福立,孙银萍,秦杰,荣建胜. DC-CIK细胞联合EGFR-TKI治疗35例老年晚期EGFR突变肺癌的效果[J]. 山东大学学报 (医学版), 2022, 60(7): 110-117.
[4] 李博,刘竞芳,鲍金鹏,李祥泽,秦广洋,田虎. 三维可视化联合吲哚菁绿荧光影像技术在51例原发性肝癌腹腔镜切除术中的应用价值[J]. 山东大学学报 (医学版), 2022, 60(3): 83-88.
[5] 左立平,蒋丰洋,周斌彬,范金蕾,梁永锋,邓展昊,于德新. 术前MRI在预测169例肝细胞肝癌微血管侵犯及早期复发的价值[J]. 山东大学学报 (医学版), 2022, 60(3): 89-95.
[6] 柴佳威,朱坤兵,张华伟,崔凤云,王甜甜,田兴松. 1例肝细胞肝癌甲状腺转移罕见病例报道与文献回顾[J]. 山东大学学报 (医学版), 2021, 59(3): 120-124.
[7] 孙薏丰,高玉,梁永媛,高杨. CPLX2在30例肝癌组织的表达及其对体外细胞增殖与侵袭的影响[J]. 山东大学学报 (医学版), 2020, 1(9): 34-39.
[8] 邵倩倩,王景溥,王庆杰. 抗凋亡转录因子在肝细胞肝癌中的表达及预后作用[J]. 山东大学学报 (医学版), 2018, 56(12): 7-12.
[9] 杨飞龙,周尊林,任巨超,闫磊,刘海南, 张温花,俞能旺,李大伟,徐忠华. 肝癌衍生生长因子对前列腺癌细胞增殖的影响及其机制[J]. 山东大学学报 (医学版), 2018, 56(1): 62-69.
[10] 王宁宁,曲琳莉,曹芳丽,梁磊. CT矢状位重建在膈顶部肝肿瘤微波消融治疗中的应用[J]. 山东大学学报（医学版）, 2017, 55(8): 71-75.
[11] 贺武斌,苏荣健. 白头翁皂苷D联合索拉非尼对人肝癌细胞侵袭与转移的影响[J]. 山东大学学报（医学版）, 2016, 54(7): 18-22.
[12] 岳倩倩,王新怡,杨志强,姜舒. 富血供肝脏转移瘤与肝细胞肝癌的能谱CT成像定量分析[J]. 山东大学学报（医学版）, 2016, 54(7): 50-55.
[13] 宗兆运,李霞,韩振龙,王显腾,郭春,张利宁,石永玉. 肿瘤相关巨噬细胞对肝癌细胞c-Met分子表达的影响[J]. 山东大学学报（医学版）, 2016, 54(3): 14-18.
[14] 李雪峰, 王双龙, 于美, 崔红, 孙媛媛, 董磊. 经皮经肝超声造影显像评估兔VX2肝癌腹腔淋巴结转移的实验研究[J]. 山东大学学报（医学版）, 2015, 53(6): 13-17.
[15] 谷旭, 王夏青, 任万华, 秦成勇, 韩国庆. NPRL2、Survivin在肝细胞肝癌中的表达及其临床意义[J]. 山东大学学报（医学版）, 2015, 53(6): 68-72.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 2018 《山东大学学报(医学版)》编辑部　
地址:山东大学科技期刊社（济南市历城区山大南路27号山东大学中心校区明德楼B座721室） 电话: 0531-88366918 E-mail: xbyxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发