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Prostaglandin E1 protects hepatocytes against endoplasmic reticulum stress-induced apoptosis via protein kinase A-dependent induction of glucose-regulated protein 78 expression 被引量:8
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作者 Fang-Wan Yang Yu Fu +5 位作者 Ying Li Yi-Huai He Mao-Yuan Mu Qi-Chuan Liu Jun Long Shi-De Lin 《World Journal of Gastroenterology》 SCIE CAS 2017年第40期7253-7264,共12页
AIM To investigate the protective effect of prostaglandin E1(PGE1) against endoplasmic reticulum(ER) stressinduced hepatocyte apoptosis, and to explore its underlying mechanisms.METHODS Thapsigargin(TG) was used to in... AIM To investigate the protective effect of prostaglandin E1(PGE1) against endoplasmic reticulum(ER) stressinduced hepatocyte apoptosis, and to explore its underlying mechanisms.METHODS Thapsigargin(TG) was used to induce ER stress in the human hepatic cell line L02 and hepatocarcinomaderived cell line Hep G2. To evaluate the effects of PGE1 on TG-induced apoptosis, PGE1 was used an hour prior to TG treatment. Activation of unfolded protein response signaling pathways were detected by western blotting and quantitative real-time RTPCR. Apoptotic index and cell viability of L02 cells and Hep G2 cells were determined with flow cytometry and MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium] assay. RESULTS Pretreatment with 1 μmol/L PGE1 protected against TG-induced apoptosis in both L02 cells and Hep G2 cells. PGE1 enhanced the TG-induced expression of C/EBP homologous protein(CHOP), glucose-regulated protein(GRP) 78 and spliced X box-binding protein 1 at 6 h. However, it attenuated their expressions after 24 h. PGE1 alone induced protein and m RNA expressions of GRP78; PGE1 also induced protein expression of DNA damage-inducible gene 34 and inhibited the expressions of phospho-PKR-like ER kinase, phosphoeukaryotic initiation factor 2α and CHOP. Treatment with protein kinase A(PKA)-inhibitor H89 or KT5720 blocked PGE1-induced up-regulation of GRP78. Further, the cytoprotective effect of PGE1 on hepatocytes was not observed after blockade of GRP78 expression by H89 or small interfering RNA specifically targeted against human GRP78.CONCLUSION Our study demonstrates that PGE1 protects against ER stress-induced hepatocyte apoptosis via PKA pathwaydependent induction of GRP78 expression. 展开更多
关键词 HEPATOCYTES Endoplasmic reticulum stress THAPSIGARGIN glucose-regulated protein 78 protein kinase A APOPTOSIS
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Role of Glucose-regulated Protein 78 in Early Brain Injury after Experimental Subarachnoid Hemorrhage in Rats 被引量:4
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作者 刘祺 赵冬 +4 位作者 姬云翔 黄啸元 杨鹏 王业忠 雷霆 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2016年第2期168-173,共6页
Early brain injury(EBI) plays a key role in the pathogenesis of subarachnoid hemorrhage(SAH). This study investigated the role of glucose-regulated protein 78(GRP78) in EBI after SAH. Male Sprague-Dawley rats(n... Early brain injury(EBI) plays a key role in the pathogenesis of subarachnoid hemorrhage(SAH). This study investigated the role of glucose-regulated protein 78(GRP78) in EBI after SAH. Male Sprague-Dawley rats(n=108) weighing 260±40 g were divided into control, sham-operated, and operated groups. Blood was injected into the prechiasmatic cistern of rats in the operated group. Neurological scores, ultrastructures of neurons, apoptosis, and GRP78 expression in the hippocampus were examined using Garcia scoring system, transmission electron microscopy, terminal deoxynucleotidyl transferase-mediated d UTP nick-end labelling, and Western blotting at 1, 6, 12, 24, 48, and 72 h after SAH, respectively. The results showed that neurological scores were significantly decreased in the operated group as compared with those in control and sham-operated groups at 12, 24, 48, and 72 h. Metachromatin, chromatin pyknosis at the edge, endoplasmic reticulum swelling, and invagination of nuclear membrane were observed at 24 h in the operated group, indicating the early morphological changes of apoptosis. The number of apoptotic cells was significantly increased in the operated group as compared with that in control and sham-operated groups at 6, 12, 24, 48, and 72 h. The GRP78 protein expression levels in the operated group were significantly elevated at all time points and reached the peak at 12 h. GRP78 expression was positively associated with apoptosis cells and negatively with neurological scores. In conclusion, EBI was demonstrated to occur after SAH and GRP78 was involved in the development of EBI after SAH. 展开更多
关键词 early brain injury subarachnoid hemorrhage glucose-regulated protein 78 endoplasmic reticulum stress
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Effects of serum containing natural cerebrolysin on glucose-regulated protein 78 and CCAAT enhancer-binding protein homologous protein expression in neuronal PC12 cells following tunicamycin-induced endoplasmic reticulum stress 被引量:5
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作者 Zhengzhi Wu Ming Li +3 位作者 Andrew C.J. Huang O Xiuqing Jia Yinghong Li Manyin Chen 《Neural Regeneration Research》 SCIE CAS CSCD 2009年第2期92-97,共6页
BACKGROUND: Glucose-regulated protein 78 (GRP78), a marker of endoplasmic reticulum stress, can prolong cell survival. Alternatively, CCAAT enhancer-binding protein homologous protein (CHOP), a transcription fact... BACKGROUND: Glucose-regulated protein 78 (GRP78), a marker of endoplasmic reticulum stress, can prolong cell survival. Alternatively, CCAAT enhancer-binding protein homologous protein (CHOP), a transcription factor specific for endoplasmic reticulum stress, can cause cell cycle arrest and cell apoptosis. OBJECTIVE: To study the protective effects of serum containing natural cerebrolysin on endoplasmic reticulum stress in tunicamycin-induced neuronal PC12 cells, and analyze the influence on GRP78 and CHOP expressions. DESIGN, TIME AND SETTING: A parallel controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine, Shenzhen Hospital, Southern Medical University, between March 2006 and August 2008. MATERIALS: Adult Sprague-Dawley rats were perfused with natural Cerebrolysin aqueous extract (0.185 g/kg/d) to produce serum containing natural Cerebrolysin. Physiological saline was used to produce blank serum. PC12 cell line was provided by Shanghai Institute of Cell Biology, Chinese Academy of Science. Tunicamycin was provided by Sigma (St. Louis, USA), and natural Cerebrolysin, containing ginseng, rhizoma gastrodiae, and gingko leaf (1:2:2), by Shengzhen Institute of Integrated Western and Traditional Chinese Medicine. METHODS: PC12 cells were treated with DMEM culture media containing 10% blank serum (normal control group), tunicamycin (1 μg/mL; model group), and 5%, 10%, and 15% serum containing natural cerebrolysin and tunicamycin (1 μ g/mL; low-, moderate-, and high-dose serum containing natural cerebrotysin groups), for 2 hours. MAIN OUTCOME MEASURES: PC12 cells were treated with tunicamycin for 48 hours after which apoptosis was measured using the TUNEL method to calculate apoptotic index. GRP78 expression was detected using immunocytochemistry. After 24 hours of treatment with tunicamycin, GRP78 and CHOP mRNA expressions were measured using RT-PCR. RESULTS: The apoptotic index and CHOP mRNA expression were in the model group and three cerebrolysin groups were significantly increased when compared to the normal control group (P 〈 0.05). In contrast, GRP78 mRNA and protein expressions were significantly decreased (P 〈 0.05). CONCLUSION: Serum containing natural cerebrolysin significantly reduced apoptosis in neuronal PC12 cells following tunicamycin-induced endoplasmic reticulum stress. These results may be related to an up-regulation of GRP78 expression and down-regulation of CHOP expression, both of which displayed dose-dependent effects. 展开更多
关键词 natural cerebrolysin PC12 cell strain endoplasmic reticulum stress TUNICAMYCIN glucose regulated protein 78 CCAAT/enhancer-binding protein homologous protein
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Clinical implications of forkhead box M1, cyclooxygenase-2, and glucose-regulated protein 78 in breast invasive ductal carcinoma
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作者 Jie Bai Ying Li Li Cai 《World Journal of Clinical Cases》 SCIE 2023年第30期7284-7293,共10页
BACKGROUND Breast infiltrating ductal carcinoma(BIDC)represents the largest heterotypic tumor group,and an in-depth understanding of the pathogenesis of BIDC is key to improving its prognosis.AIM To analyze the expres... BACKGROUND Breast infiltrating ductal carcinoma(BIDC)represents the largest heterotypic tumor group,and an in-depth understanding of the pathogenesis of BIDC is key to improving its prognosis.AIM To analyze the expression profiles and clinical implications of forkhead box M1(FOXM1),cyclooxygenase-2(COX-2),and glucose-regulated protein 78(GRP78)in BIDC.METHODS A total of 65 BIDC patients and 70 healthy controls who presented to our hospital between August 2019 and May 2021 were selected for analysis.The peripheral blood FOXM1,COX-2,and GRP78 levels in both groups were measured and the association between their expression profiles in BIDC was examined.Additionally,we investigated the diagnostic value of FOXM1,COX-2,and GRP78 in patients with BIDC and their correlations with clinicopathological features.Furthermore,BIDC patients were followed for 1 year to identify factors influencing patient prognosis.RESULTS The levels of FOXM1,COX-2,and GRP78 were significantly higher in BIDC patients compared to healthy controls(P<0.05),and a positive correlation was observed among them(P<0.05).Receiver operating characteristic analysis demonstrated that FOXM1,COX-2,and GRP78 had excellent diagnostic value in predicting the occurrence of BIDC(P<0.05).Subsequently,we found significant differences in FOXM1,COX-2,and GRP78 levels among patients with different histological grades and metastasis statuses(with vs without)(P<0.05).Cox analysis revealed that FOXM1,COX-2,GRP78,increased histological grade,and the presence of tumor metastasis were independent risk factors for prognostic death in BIDC(P<0.001).CONCLUSION FOXM1,COX-2,and GRP78 exhibit abnormally high expression in BIDC,promoting malignant tumor development and closely correlating with prognosis.These findings hold significant research implications for the future diagnosis and treatment of BIDC. 展开更多
关键词 Diagnostic value Forkhead box M1 CYCLOOXYGENASE-2 glucose-regulated protein 78 Clinical implications
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Glucose-regulated protein 78 inhibits scavenger receptor A-mediated internalization of acetylated low density lipoprotein
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作者 Ben, Jingjing Gao, Song +8 位作者 Zhu, Xudong Zheng, Yuan Zhuang, Yan Bai, Hui Xu, Yong Ji, Yong Sha, Jiahao He, Zhigang Chen, Qi 《南京医科大学学报(自然科学版)》 CAS CSCD 北大核心 2010年第1期105-105,共1页
Class A scavenger receptor(SR-A) plays an important role in foam cell formation.However, the mechanism underlying the internalization of the receptor-ligand complexes remains unclear.The aim of the present study was t... Class A scavenger receptor(SR-A) plays an important role in foam cell formation.However, the mechanism underlying the internalization of the receptor-ligand complexes remains unclear.The aim of the present study was to investigate the molecular mechanism to regulate SR-A-mediated intracellular lipid accumulation in macrophages A pull-clown assay was performed and glucoseregulated protein 78(GRP78) was identified to bind with the cytoplasmic domain of SR-A(CSR-A).Immunoprecipitation and artificially expressed protein binding assay demonstrated the direct specific binding of GRP78 with SR-A in cells.Indirect immunofluorescence assay and western blot analysis showed their co-localization in membrane and cytoplasm.Over-expression of GRP78 specifically inhibited SR-A-mediated uptake of fluorescent acetylated low-density lipoprotein, a specific ligand for SR-A, without altering cellular SR-A expression and binding ability, and significantly inhibited cholesterol ester accumulation in cells, which can be partly attributed to the suppression of c-Jun-NH2-terminal kinase signaling pathway.These results suggest that GRP78 may act as an inhibitor of SR-A-mediated internalization of modified low-density lipoprotein into macrophages(C) 2009 Elsevier Inc.All rights reserved. 展开更多
关键词 受体 泡沫细胞 免疫沉淀 蛋白质
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Correlation between clinicopathology and expression of heat shock protein 70 and glucose-regulated protein 94 in human colonic adenocarcinoma 被引量:39
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作者 Xiao-PingWang Fan-RongQiu +1 位作者 Guo-ZhenLiu Rui-FenChen 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第7期1056-1059,共4页
AIM: To investigate the correlation between clinicopathology and expression of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human colonic carcinoma. METHODS: The expression of HSP70 and gr... AIM: To investigate the correlation between clinicopathology and expression of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human colonic carcinoma. METHODS: The expression of HSP70 and grp94 was studied in 80 human colonic cancers with or without metastasis as well as in their adjacent mucous membrane by way of immunohistochemistry and pathology photograph analysis. RESULTS: The expression of HSP70 and grp94 was significantly higher in cancer than that in adjacent mucous membrane (92.5%, 85.0% vs 56.3%, 42.5%, P<0.01). HSP70 and grp94 expressed higher in moderately- and poorly-differentiated colonic cancers than that in their adjacent tissues (93.7%, 87.5%; 100%, 90% vs56.3%, 42.5%;P<0.01). Dukes C and D stages of colonic cancers showed higher positive rates than Dukes A and B stage groups (97.1%, 91.2%; 100%, 90.9%; vs 80%, 70%; 78.6%, 71.4%; P<0.05). There were definite differences in HSP70 and grp94 expression between metastasis groups and non-metastasis groups (100% vs 75%, 100% CONCLUSION: The HSP70 and grp94 expression rates in colonic cancer groups are significantly higher than that in their adjacent mucous membrane. The HSP70 and grp94 expression in poorly-differentiated colonic cancers with metastasis is significantly higher than well-differentiated cancers without metastasis. The overexpression of HSP70 and grp94 can be used as diagnostic or prognostic markers for colonic cancer. 展开更多
关键词 Heat shock protein 70 glucose-regulated protein 94 Colonic adenocarcinoma
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Expression and significance of heat shock protein 70 and glucose-regulated protein 94 in human esophageal carcinoma 被引量:28
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作者 Xiao-PingWang Guo-ZhenLiu Ai-LiSong Rui-FenChen Hai-YanLi YuLiu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第3期429-432,共4页
AIM: To investigate the expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human esophageai carcinoma and adjacent normal tissues. METHODS: The expression of HSP70... AIM: To investigate the expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human esophageai carcinoma and adjacent normal tissues. METHODS: The expression of HSP70 and grp94 in 78 human esophageai cancer and adjacent normal tissues was studied by immunohistochemistry and pathology photograph analysis. RESULTS: Both esophageai cancer and adjacent normal tissues could express HSP70 and grp94. Of the 78 cases of esophageai carcinoma, 95.0%(72/78) showed positive HSP70, mainly stained in nuclei, while grp94 was mainly stained in cell plasma, and the positive rate was 71.8% (56/78).There was a significant difference in the expression of HSP70 and grp94 between esophageai cancer and adjacent normal tissues (P<0.01). Compared with adjacent normal tissues, there was a significant difference between differential types and HSP70 expression (P<0.01). CONCLUSION: HSP70 and grp94 express differently in cell plasma and nuclei. The expression intensity of HSP70 is related to the differentiation of esophageai carcinoma. 展开更多
关键词 Esophageal carcinoma Heat shock protein 70 glucose-regulated protein 94
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Co-expression of heat shock protein 70 and glucose-regulated protein 94 in human gastric carcinoma cell line BGC-823 被引量:9
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作者 Xiao-PingWang JingLiao +2 位作者 Guo-ZhenLiu Xing-CuiWang Hong-WeiShang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第23期3601-3604,共4页
AIM:To investigate the co-expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human gastric carcinoma cell line BGC-823. METHODS: The expression and localization of... AIM:To investigate the co-expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human gastric carcinoma cell line BGC-823. METHODS: The expression and localization of HSP70 and grp94 in human gastric carcinoma cell line BGC-823 were determined by immunocytochemistry and indirect immunofluorescence cytochemical staining. Flow cytometry was used to analyze the correlation between expression of HSP70, grp94 and cell cycle in BGC-823 cell line. RESULTS: Gastric cancer cell line BGC-823 expressed high level of HSP70 and grp94. The positive rate of HSP70 and grp94 was 84.9±4.94% and 79.6±5.16%, respectively. Both of them were stained in cell plasma. There was a significant difference compared with control group (1.9±0.94%, P<0.01). During the cell cycle, HSP70 and grp94 were continuously expressed in BGC-823. CONCLUSION: HSP70 and grp94 are highly expressed in human gastric carcinoma BGC-823 cells through the whole cell cycle. There is no relationship between expression of HSP70, grp94 and cell cycle. 展开更多
关键词 Heat shock protein 70 glucose-regulated protein 94 Gastric carcinoma
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血清FSTL1、GRP78对老年COPD患者疾病严重程度及再入院的评估价值
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作者 李亚男 夏君燕 +1 位作者 连雪梅 高永红 《国际检验医学杂志》 2025年第1期44-48,共5页
目的探讨血清卵泡抑素样蛋白1(FSTL1)、葡萄糖调节蛋白78(GRP78)对老年慢性阻塞性肺疾病(COPD)患者疾病严重程度及再入院的评估价值。方法选取该院于2020年3月至2023年5月收治的100例老年COPD患者作为COPD组,根据病情严重程度分为Ⅰ级(3... 目的探讨血清卵泡抑素样蛋白1(FSTL1)、葡萄糖调节蛋白78(GRP78)对老年慢性阻塞性肺疾病(COPD)患者疾病严重程度及再入院的评估价值。方法选取该院于2020年3月至2023年5月收治的100例老年COPD患者作为COPD组,根据病情严重程度分为Ⅰ级(35例)、Ⅱ级(46例)、Ⅲ级(19例)。另选取同期体检的100例老年志愿者作为对照组。采用酶联免疫吸附试验(ELISA)检测两组血清FSTL1、GRP78水平。采用Spearman和Pearson相关性分析血清FSTL1、GRP78水平与一般资料的相关性,采用受试者工作特征(ROC)曲线分析COPD患者血清FSTL1、GRP78水平对疾病严重程度及再入院的评估价值。结果与对照组相比,COPD组血清FSTL1、GRP78水平均显著升高(P<0.05)。COPD患者血清FSTL1、GRP78水平随着病情严重程度增加而依次升高(P<0.05)。与非再入院组相比,再入院组患者血清FSTL1、GRP78水平显著升高(P<0.05)。Spearman相关性分析显示,血清FSTL1、GRP78水平与吸烟史、高血压史、病情严重程度呈正相关(P<0.05);Pearson相关性分析显示,血清FSTL1、GRP78水平与动脉血氧分压(PaO 2)、第1秒用力呼吸容积与用力肺活量的比值(FEV 1/FVC)、第1秒用力呼气容积占预计值百分比(FEV 1%pred)呈负相关(P<0.05),与二氧化碳分压(PaCO 2)呈正相关(P<0.05)。ROC曲线分析显示,COPD患者血清FSTL1、GRP78及二者联合评估病情严重程度及再入院的曲线下面积(AUC)均较高,并且二者联合明显均优于单独评估(P<0.05)。结论COPD患者血清FSTL1、GRP78水平均明显升高,二者与病情严重程度相关,对病情严重程度及再入院的评估价值较高。 展开更多
关键词 老年慢性阻塞性肺疾病 卵泡抑素样蛋白1 葡萄糖调节蛋白78 再入院
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血清FABP4、GRP78及FOXO1与急性脑梗死患者预后的相关性
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作者 任晓飞 刘娜 +2 位作者 耿颖 甄运钰 芮淑红 《检验医学与临床》 2025年第3期304-308,共5页
目的探讨脂肪酸结合蛋白4(FABP4)、葡萄糖调节蛋白78(GRP78)、叉头框转录因子O亚族1(FOXO1)与急性脑梗死(ACI)患者预后的相关性。方法选取2021年6月至2023年1月该院收治的148例ACI患者(ACI组)作为研究对象,另选取148例同期于该院进行体... 目的探讨脂肪酸结合蛋白4(FABP4)、葡萄糖调节蛋白78(GRP78)、叉头框转录因子O亚族1(FOXO1)与急性脑梗死(ACI)患者预后的相关性。方法选取2021年6月至2023年1月该院收治的148例ACI患者(ACI组)作为研究对象,另选取148例同期于该院进行体检的健康体检者作为对照组。根据ACI患者出院后3个月的预后情况将其分为预后良好组和预后不良组;比较各组血清FABP4、GRP78、FOXO1水平;采用多因素Logistic回归分析ACI患者预后不良的影响因素。绘制受试者工作特征(ROC)曲线分析血清FABP4、GRP78、FOXO1单独及联合检测对ACI患者预后不良的预测价值。结果ACI组血清FABP4、GRP78水平均高于对照组,FOXO1水平低于对照组,差异均有统计学意义(P<0.05)。预后良好组纳入92例,预后不良组纳入56例。预后不良组年龄、梗死灶面积均大于预后良好组,FABP4、GRP78水平均高于预后良好组,FOXO1水平低于预后良好组,差异均有统计学意义(P<0.05)。多因素Logistic回归分析结果显示,FABP4、GRP78、FOXO1、年龄、梗死灶面积是ACI患者预后不良的影响因素(P<0.05)。ROC曲线分析结果显示,血清FABP4、GRP78、FOXO1单独及联合预测ACI患者预后不良的曲线下面积(AUC)分别为0.795、0.819、0.784、0.927;血清FABP4、GRP78、FOXO13项联合预测ACI患者预后不良的AUC优于各自单独预测的AUC(Z联合检测-FABP4=3.909,Z联合检测-GRP78=3.171,Z联合检测-FOXO1=4.494,P<0.05)。结论ACI患者血清FABP4、GRP78水平均升高,FOXO1水平降低,3项联合检测对ACI患者预后有较高的预测价值。 展开更多
关键词 脂肪酸结合蛋白4 葡萄糖调节蛋白78 叉头框转录因子O亚族1 急性脑梗死 预后
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血清IL-33、GRP78、vWF对急性大出血并发输血相关性急性肺损伤患者预后的预测价值
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作者 朱晶沐 单来友 +2 位作者 陈治宇 孟鹏飞 贾旭颖 《临床肺科杂志》 2025年第2期209-214,共6页
目的观察血清白细胞介素-33(IL-33)、葡萄糖调节蛋白78(GRP78)、血管性血友病因子(vWF)的表达水平,并分析其对急性大出血并发输血相关性急性肺损伤(TRALI)患者预后的价值。方法收集本院2021年1月—2023年5月收治的急性大出血并发TRALI... 目的观察血清白细胞介素-33(IL-33)、葡萄糖调节蛋白78(GRP78)、血管性血友病因子(vWF)的表达水平,并分析其对急性大出血并发输血相关性急性肺损伤(TRALI)患者预后的价值。方法收集本院2021年1月—2023年5月收治的急性大出血并发TRALI患者为TRALI组(n=90),对照组为同期在本院收治的未发生TRALI的急性大出血患者(n=86)。根据TRALI患者10天内的生存情况,分为生存组(n=79)和死亡组(n=11)。血清IL-33,GRP78水平检测用ELISA法;血清vWF水平检测用免疫比浊法;TRALI患者血清中IL-33、GRP78、vWF水平相关性分析用Pearson法;比较生存组和死亡组患者的临床资料;Logistic回归分析影响TRALI患者预后的相关因素;ROC曲线评估血清IL-33、GRP78、vWF水平对TRALI患者预后的预测价值。结果与对照组相比,TRALI组血清IL-33、GRP78、vWF水平均升高(P<0.05)。与生存组相比,死亡组血清IL-33、GRP78、vWF水平均升高(P<0.05)。TRALI组血清IL-33与GRP78、vWF水平以及GRP78与vWF水平间均呈正相关(r=0.516、0.495、0.453,P<0.001)。生存组和死亡组患者是否有过敏史、既往输血史以及输血次数比较,差异有统计学意义(P<0.05),且过敏史、既往输血史、输血次数、血清IL-33、GRP78、vWF水平是TRALI患者预后死亡的影响因素(P<0.05)。血清IL-33、GRP78、vWF三者联合预测TRALI患者预后的AUC为0.977(95%CI:0.921~0.997),灵敏度和特异度分别为90.91%和89.87%,三者联合预测优于单独预测(Z_(三者联合-IL-33)=2.056,P=0.040;Z_(三者联合-GRP78)=3.101,P=0.002;Z_(三者联合-vWF)=2.027,P=0.043)。结论在急性大出血并发TRALI患者中血清IL-33、GRP78、vWF水平均升高,三者联合检测对急性大出血并发TRALI患者预后具有较高的临床预测价值。 展开更多
关键词 白细胞介素-33 葡萄糖调节蛋白78 血管性血友病因子 急性大出血 输血相关性急性肺损伤 预后
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Transformation of Arabidopsis by Rice OsWRKY78::GFP Fusion Gene and Subcellular Localization of OsWRKY78 Protein 被引量:1
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作者 刘顺枝 张美 +1 位作者 唐馨 王小兰 《Agricultural Science & Technology》 CAS 2012年第7期1395-1398,共4页
[Objective] The study was to understand the subcellular localization of OsWRKY78 protein in plants. [Method] Primers specific for OsWRKY78 gene were designed according to the OsWRKY78 full length sequence in Genbank. ... [Objective] The study was to understand the subcellular localization of OsWRKY78 protein in plants. [Method] Primers specific for OsWRKY78 gene were designed according to the OsWRKY78 full length sequence in Genbank. The gene was cloned by RT-PCR method. The gene was then recombined into a plasmid expression vector carrying green fluorescent protein (GFP) gene, pBinGFP. The recombinant was confirmed by PCR and enzyme digestion. The recombinant plasmid pBinGFP-OsWRKY was transformed into Arabidopsis through Agrobacterium tumefaciens strain GV3101 and transgenic plants were obtained. [Result] Measured by fluorescence microscopy, the expression of OsWRKY78 and GFP fusion protein in root tip cells was localized in the nucleus. [Conclusion] This study laid the foundation for further investigating the function of OsWRKY78 gene and its role in related signal transduction and provided theoretical basis for exploring the relation between OsWRKY78 gene and brown planthoppers. 展开更多
关键词 OsWRKY78 GENE Green fluorescent protein GENE Expression vector SUBCELLULAR localization
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绝经后骨质疏松症患者外周血中GRP78和ATF4的表达水平及临床意义
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作者 田利军 郭宇松 +1 位作者 王晓燕 郭哲铭 《空军航空医学》 2024年第1期42-46,共5页
目的探究绝经后骨质疏松症(postmenopausal osteoporosis,PMO)患者外周血中葡萄糖调节蛋白78(glucoseregulated protein 78,GRP78)和激活转录因子4(activating transcription factor 4,ATF4)的水平及临床意义。方法选取2018年12月—2021... 目的探究绝经后骨质疏松症(postmenopausal osteoporosis,PMO)患者外周血中葡萄糖调节蛋白78(glucoseregulated protein 78,GRP78)和激活转录因子4(activating transcription factor 4,ATF4)的水平及临床意义。方法选取2018年12月—2021年5月石家庄市第三医院创伤骨科收治的96例PMO患者为PMO组,另选取同期96名体检健康的绝经女性为正常组。比较PM O组和正常组的一般资料;采用酶联免疫吸附法测定血清GR P78、ATF4水平;利用电化学发光免疫分析仪测定血清β-胶原特殊序列(β-Crosslaps)、Ⅰ型前胶原氨基端前肽(procollagenⅠtype N-terminal propeptide,PINP)水平;利用双能X线骨密度仪检测腰椎骨密度(bone mineral density,BMD);采用Pearson法分析PMO患者血清GRP78、ATF4水平与β-Crosslaps、PIN P、BM D的相关性及PMO患者血清GR P78表达水平与ATF4的相关性;用Logistic回归分析PMO发生的影响因素。结果PMO组患者腰臀比、BMD低于正常组(t=9.76、15.47,P均<0.001);血清GRP78、ATF4、β-Crosslaps、PINP水平高于正常组(t=18.32、16.81、8.15、14.84,P均<0.001);PMO患者血清GRP78、ATF4水平与β-Crosslaps、PINP均呈正相关(r=0.56、0.48、0.48、0.52,P<0.001、<0.001、<0.001、=0.006),与BMD呈负相关(r=-0.54、-0.44,P均<0.001);PMO患者血清GRP78水平与ATF4呈正相关(r=0.595,P<0.001);PINP、GRP78、β-Crosslaps、ATF4是影响PMO发生的危险因素(OR=2.518、2.672、2.271、2.336,P均<0.001),BMD是影响PMO发生的保护因素(OR=0.583,P<0.001)。结论PMO患者血清GRP78、ATF4水平较高,均与β-Crosslaps、PINP、BMD相关,GRP78、ATF4可能是诊治PMO的潜在靶标,测定血清GRP78、ATF4水平有助于临床防治PMO。 展开更多
关键词 葡萄糖调节蛋白78 绝经后骨质疏松症 β-胶原特殊序列 激活转录因子4 Ⅰ型前胶原氨基端前肽
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血清ADAM15、GRP78联合CT肺动脉造影对急性肺栓塞的诊断价值
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作者 马玉萍 栾丽 陶思冥 《中国CT和MRI杂志》 2024年第8期47-50,共4页
目的探究血清整合素金属蛋白酶15(ADAM15)、葡萄糖调节蛋白78(GRP78)联合CT肺动脉造影(CTPA)对急性肺栓塞(APE)的诊断价值。方法选取本院2021年6月至2023年6月收治的86例疑似APE患者,患者均行CTPA检查;根据病情严重程度将APE患者分为中... 目的探究血清整合素金属蛋白酶15(ADAM15)、葡萄糖调节蛋白78(GRP78)联合CT肺动脉造影(CTPA)对急性肺栓塞(APE)的诊断价值。方法选取本院2021年6月至2023年6月收治的86例疑似APE患者,患者均行CTPA检查;根据病情严重程度将APE患者分为中高危组和低危组;酶联免疫吸附法检测ADAM15、GRP78水平;Pearson相关性分析血清ADAM15、GRP78与CTPA指标的相关性;中高危APE的影响因素采用多因素Logistic回归分析;绘制ROC曲线分析血清ADAM15、GRP78对中高危APE的诊断价值。结果86例患者经过CTPA检测出栓子702个,86例患者在不同肺动脉部位表现出充盈缺损等,病变部位主要位于双肺29例,左肺30例,右肺27例。4种栓塞类型42例中心型,98例偏心型,26例附壁血栓型,30例完全堵塞型。中高危组RVD/LVD、RV-LD/LV-LD、Qanadli栓塞指数显著高于低危组(P<0.05)。中高危组血清ADAM15、GRP78水平显著高于低危组(P<0.05)。根据Pearson相关性分析得知,血清ADAM15与GRP78呈正相关(P<0.05),二者均与RVD/LVD、RVLD/LV-LD、Qanadli栓塞指数呈正相关(P<0.05)。多因素Logistic回归分析得知ADAM15、GRP78、RVD/LVD、RV-LD/LV-LD、Qanadli栓塞指数为影响中高危APE患者的危险因素(P<0.05)。根据ROC曲线得知,血清ADAM15、GRP78、RVD/LVD、RVLD/LV-LD和Qanadli栓塞指数五者联合诊断中高危APE的AUC为0.990,五者联合优于各自单独诊断(Z_(联合vs ADAM15)=2.691、Z_(联合vs GRP78)=2.578、Z_(联合vs RVD/LVD)=2.710、Z_(联合vs RV-LD/LV-LD)=2.714、Z联合vs Qanadli栓塞指数=2.698,P均<0.05)。结论血清ADAM15、GRP78在APE患者中显著升高,二者联合CTPA可提高对APE的诊断价值。 展开更多
关键词 整合素金属蛋白酶15 葡萄糖调节蛋白78 CT肺动脉造影 急性肺栓塞 诊断
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天香丹通过GRP78/NLRP3信号通路调控内质网应激抗动脉粥样硬化的作用机制研究 被引量:2
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作者 张夏夏 白银雪 +3 位作者 吴丹丹 孙龙飞 任珊 安冬青 《中西医结合心脑血管病杂志》 2024年第11期1955-1960,共6页
目的:基于葡萄糖调节蛋白78(GRP78)/NOD样受体蛋白3(NLRP3)信号通路探讨天香丹通过调控内质网应激抗动脉粥样硬化的作用机制。方法:将雄性载脂蛋白E基因敲除(ApoE^(-/-))小鼠随机分为模型组、阿托伐他汀组、天香丹组,每组11只,给予高脂... 目的:基于葡萄糖调节蛋白78(GRP78)/NOD样受体蛋白3(NLRP3)信号通路探讨天香丹通过调控内质网应激抗动脉粥样硬化的作用机制。方法:将雄性载脂蛋白E基因敲除(ApoE^(-/-))小鼠随机分为模型组、阿托伐他汀组、天香丹组,每组11只,给予高脂饲料诱导动脉粥样硬化模型;另取10只雄性C57BL/6J小鼠作为正常组。于造模10周后给予相应药物治疗,连续干预12周。采用苏木精-伊红(HE)染色观法察胸主动脉病理特点;酶联免疫吸附试验(ELISA)检测血清白细胞介素1β(IL-1β)、白细胞介素18(IL-18)、活性氧(ROS)含量;免疫组织化学法(IHC-P)检测胸主动脉中GRP78和NLRP3蛋白阳性表达水平。结果:各实验组存活6只。与模型组比较,阿托伐他汀组和天香丹组IL-1β、IL-18、ROS表达含量均降低(P<0.05或P<0.01);GRP78、NLRP3蛋白阳性表达均明显减少(P<0.05或P<0.01)。结论:天香丹能有效改善动脉粥样硬化性心血管疾病小鼠炎症反应,其作用机制可能与调控内质网应激GRP78/NLRP3信号通路抑制炎症小体活化、减轻动脉粥样硬化炎症损伤有关。 展开更多
关键词 动脉粥样硬化 天香丹 内质网应激 葡萄糖调节蛋白78 NOD样受体蛋白3 实验研究
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流体剪应力对人脐静脉内皮细胞葡萄糖调节蛋白78和C/EBP同源蛋白表达的影响
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作者 张森 王先伟 +3 位作者 黄家明 张昊然 李梅 陈东 《中国脑血管病杂志》 CAS CSCD 北大核心 2024年第6期388-395,共8页
目的探索流体剪应力对人脐静脉内皮细胞(HUVECs)葡萄糖调节蛋白78(GRP78)和C/EBP同源蛋白(CHOP)表达的影响。方法以HUVECs作为实验细胞,设计并构建流体动力学模拟实验系统,控制流体动力学模拟实验系统中灌流液的流速,以实现对实验细胞... 目的探索流体剪应力对人脐静脉内皮细胞(HUVECs)葡萄糖调节蛋白78(GRP78)和C/EBP同源蛋白(CHOP)表达的影响。方法以HUVECs作为实验细胞,设计并构建流体动力学模拟实验系统,控制流体动力学模拟实验系统中灌流液的流速,以实现对实验细胞施加不同的流体剪应力。按实验细胞在实验系统中所承受的不同流体剪应力,将实验细胞分为低剪应力组(A组;0.4 Pa)、中剪应力组(B组;0.8 Pa)和高剪应力组(C组;1.2 Pa)。每组HUVECs包含3个细胞玻片,每个玻片经实验系统灌流液反复循环流经12 h。采用蛋白质印迹法对各组细胞中GRP78和CHOP蛋白水平进行检测,采用实时荧光定量逆转录聚合酶链反应技术测定各组细胞GRP78和CHOP蛋白及其信使RNA(mRNA)相对水平。应用GraphPad Prism 8.0软件对数据进行统计学分析。结果(1)A、B、C组HUVECs中GRP78蛋白相对表达水平分别为1.33±0.46、0.93±0.34、0.64±0.30;多组间比较差异有统计学意义(F=36.17,P<0.05)。A组GRP78蛋白相对表达水平高于B组、C组(均P<0.01),B组GRP78蛋白相对表达水平高于C组(P=0.0013)。3组HUVECs中CHOP蛋白相对表达水平分别为:A组1.29±0.38,B组0.90±0.34,C组0.59±0.29;多组间比较差异有统计学意义(F=41.27,P<0.05)。A组CHOP蛋白相对表达水平高于B组、C组(均P<0.01),B组CHOP蛋白相对表达水平高于C组(P=0.0004)。(2)A、B、C组HUVECs中GRP78 mRNA相对表达水平分别为18.3±3.4、11.3±1.8、5.4±2.2;多组间比较差异有统计学意义(F=189.20,P<0.05)。A组GRP78 mRNA相对表达水平高于B组、C组(均P<0.01),B组GRP78 mRNA相对表达水平高于C组(P<0.01)。3组HUVECs中CHOP mRNA相对表达水平分别为:A组20.4±3.8,B组14.2±2.1,C组7.8±1.3;多组间比较差异有统计学意义(F=171.80,P<0.05)。A组CHOP mRNA相对表达水平高于B组、C组(均P<0.01),B组CHOP mRNA相对表达水平高于C组(P<0.01)。结论低流体剪应力可能增加HUVECs中GRP78、CHOP的蛋白及其mRNA表达水平。 展开更多
关键词 血流动力学 人脐静脉内皮细胞 内质网应激 流体剪应力 葡萄糖调节蛋白78 C/EBP同源蛋白
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心力衰竭急性失代偿期患者血清CHE、GRP78表达水平及其对预后的评估价值
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作者 聂安国 聂园园 赵玉铁 《海南医学》 CAS 2024年第24期3593-3597,共5页
目的检测心力衰竭急性失代偿期(ADHF)患者血清胆碱酯酶(CHE)、葡萄糖调节蛋白78(GRP78)的表达水平,并探讨其对患者预后的评估价值。方法前瞻性选取2022年3月至2023年3月濮阳油田总医院收治的120例ADHF患者作为研究组,120例慢性心力衰竭(... 目的检测心力衰竭急性失代偿期(ADHF)患者血清胆碱酯酶(CHE)、葡萄糖调节蛋白78(GRP78)的表达水平,并探讨其对患者预后的评估价值。方法前瞻性选取2022年3月至2023年3月濮阳油田总医院收治的120例ADHF患者作为研究组,120例慢性心力衰竭(CHF)患者作为对照组。比较两组患者入院时以及研究组不同临床特征[性别、年龄、体质量指数、左心室射血分数(LVEF)、纽约心脏病协会(NYHA)分级]患者入院时的血清CHE、GRP78水平。患者入院后开始随访,出院后通过电话或微信随访,随访时间180 d。依据患者随访时的预后分为预后不良组22例和预后良好组98例,比较两组患者入院时的血清CHE、GRP78水平。应用Spearman分析ADHF患者血清CHE、GRP78水平与LVEF、NYHA分级的相关性,应用独立危险因素非条件Logistic二元回归分析确定预后影响因子,应用受试者工作特征(ROC)曲线分析血清CHE、GRP78预测ADHF患者预后不良的价值。结果研究组患者入院时的血清CHE水平为(5.41±1.20)kU/L,明显低于对照组的(7.03±1.57)kU/L,GRP78水平为(476.39±91.26)ng/mL,明显高于对照组的(409.25±80.54)ng/mL,差异均有统计学意义(P<0.05);研究组患者LVEF<40%患者入院时的血清CHE水平为(4.78±0.96)kU/L,明显低于LVEF≥40%患者的(5.83±1.22)kU/L,GRP78水平为(523.46±103.58)ng/mL,明显高于LVEF≥40%患者的(445.01±84.27)ng/mL,差异均有统计学意义(P<0.05),研究组患者NYHA分级中Ⅳ级患者入院时的血清CHE水平为(4.67±0.93)kU/L,明显低于Ⅲ级患者的(5.81±1.21)kU/L,GRP78水平为(530.72±105.64)ng/mL,明显高于Ⅲ级患者的(447.14±85.19)ng/mL,差异均有统计学意义(P<0.05);Spearman分析结果显示,ADHF患者的血清CHE水平与LVEF呈正相关(r=0.512,P<0.05),与NYHA分级呈负相关(r=-0.467,P<0.05),GRP78水平与LVEF呈负相关(r=-0.537,P<0.05),与NYHA分级呈正相关(r=0.485,P<0.05);预后不良组患者入院时的血清CHE水平为(4.53±0.93)kU/L,明显低于预后良好组的(5.61±1.18)kU/L,GRP78水平为(540.83±108.75)ng/mL,明显高于预后良好组的(461.92±89.36)ng/mL,差异均有统计学意义(P<0.05);非条件Logistic二元回归分析结果显示,入院时血清CHE、GRP78均对ADHF患者预后不良具有独立预测意义(P<0.05);ROC曲线分析结果显示,入院时血清CHE、GRP78预测ADHF患者预后不良的曲线下面积(AUC)分别为0.775(95%CI:0.690~0.846)、0.758(95%CI:0.671~0.831),两者联合预测预后不良的AUC为0.919(95%CI:0.855~0.961),与单独预测的AUC比较,联合预测的AUC明显增大(P<0.05)。结论ADHF患者入院时血清CHE水平降低、GRP78水平升高,血清CHE、GRP78联合检测在临床上可作为预测预后不良的重要指标。 展开更多
关键词 心力衰竭 急性失代偿期 胆碱酯酶 葡萄糖调节蛋白78 预后 预测
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血清葡萄糖调节蛋白78在多发性骨髓瘤病人中的表达及其预后意义
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作者 唐川 邱玲 《安徽医药》 CAS 2024年第9期1773-1777,共5页
目的探讨葡萄糖调节蛋白78(GRP78)在初诊多发性骨髓瘤(MM)病人血清中的表达,及其对病人治疗反应及预后的预测价值。方法选择2017年1月至2020年12月大竹县人民医院收治的78例MM病人为MM组,同期该院门诊就诊的62例健康成人作为对照组。收... 目的探讨葡萄糖调节蛋白78(GRP78)在初诊多发性骨髓瘤(MM)病人血清中的表达,及其对病人治疗反应及预后的预测价值。方法选择2017年1月至2020年12月大竹县人民医院收治的78例MM病人为MM组,同期该院门诊就诊的62例健康成人作为对照组。收集所有参与者的外周静脉血样本,使用酶联免疫吸附测定(ELISA)法检测血清中GRP78的表达,分析GRP78表达与临床特征及治疗反应的关系。通过受试者操作特征曲线(ROC曲线)确定血清GRP78预测MM病人预后的最佳临界值,采用Kaplan-Meier曲线与Cox比例风险模型评估血清GRP78表达与MM病人预后的关系。结果MM病人的血清GRP78表达水平显著高于对照组(Z=4.12,,P<0.001),中位表达量分别为2.49(1.23,5.13)mg/L和1.29(0.75,2.14)mg/L。与多发性骨髓瘤国际预后分期(ISS)Ⅰ、Ⅱ期病人2.05(1.09,4.40)mg/L相比,血清GRP78表达在ISSⅢ期病人2.99(1.66,5.63)mg/L显著升高(Z=2.18,P=0.030)。然而,血清GRP78表达与接受硼替佐米诱导方案的MM病人的治疗反应无关。ROC曲线显示血清GRP78预测总体生存(OS)的最佳临界值为1.68 mg/L,曲线下面积(AUC)为0.70,95%CI:(0.58,0.81),灵敏度与特异度分别为78.7%和58.4%。Kaplan-Meier曲线表明GRP78高表达(≤1.68 mg/L,n=50)组生存期明显短于低表达组(>1.68 mg/L,n=28),差异有统计学意义(χ^(2)=10.99,P=0.001)。单、多因素Cox分析揭示ISS分期[HR=2.04,95%CI:(1.11,3.76),P=0.022]、治疗反应[HR=2.42,95%CI:(1.28,4.58),P=0.007]和血清GRP78表达[HR=2.96,95%CI:(1.43,6.12),P=0.003]是MM病人的独立预后因素。结论GRP78在MM病人血清中呈现异常高表达,且与ISS分期及不良预后有关,或许是MM诊断与预后风险分层的重要标志物。 展开更多
关键词 多发性骨髓瘤 葡萄糖调节蛋白78 硼替佐米 Β2-微球蛋白 治疗反应 预后
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葡萄糖调节蛋白78在心肌梗死后纤维化中的作用及机制研究
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作者 袁航 楼钶楠 林玉平 《心电与循环》 2024年第6期579-583,588,I0003,共7页
目的探究葡萄糖调节蛋白78(GRP78)在心肌梗死后纤维化中的作用及机制。方法选择雄性C57BL/6J小鼠12只,分为观察组和对照组,各6只。观察组结扎冠脉左前降支建立急性心肌梗死小鼠模型。使用苏木精伊红染色试剂盒和马松三色染色试剂盒检测... 目的探究葡萄糖调节蛋白78(GRP78)在心肌梗死后纤维化中的作用及机制。方法选择雄性C57BL/6J小鼠12只,分为观察组和对照组,各6只。观察组结扎冠脉左前降支建立急性心肌梗死小鼠模型。使用苏木精伊红染色试剂盒和马松三色染色试剂盒检测心肌纤维化区域。使用蛋白免疫印迹(WB)法检测心肌组织中GRP78、磷酸化蛋白激酶R样内质网激酶(p-PERK)、磷酸化真核启动因子2α蛋白(p-eIF2α)和CCAAT/增强子结合蛋白同源蛋白(CHOP)相对蛋白表达量。提取心脏成纤维细胞,将心脏成纤维细胞分为血管紧张素Ⅱ(AngⅡ)组、AngⅡ组+阴性对照物(sh-NC)组、AngⅡ组+抗GRP78短发夹RNA(sh-GRP78)组、AngⅡ组+sh-GRP78+衣霉素(Tn)组及对照组。使用细胞计数试剂盒8评估心肌成纤维细胞活性。使用划痕愈合实验检测心肌成纤维细胞迁移能力。使用WB法检测心肌成纤维细胞I型胶原蛋白(CollagenⅠ)、Ⅲ型胶原蛋白(CollagenⅢ)、α-平滑肌肌动蛋白(α-SMA)、GRP78、p-PERK、p-eIF2α和CHOP相对蛋白表达量。结果与对照组比较,观察组小鼠心脏组织出现明显损伤及明显纤维化,GRP78、p-PERK、p-eIF2α和CHOP相对蛋白表达量较高(均P<0.001)。在体外细胞模型中,与对照组比较,AngⅡ组心脏成纤维细胞活性较强,迁移能力较强,CollagenⅠ、CollagenⅢ、α-SMA、GRP78、p-PERK、p-eIF2α和CHOP相对蛋白表达量较高(均P<0.05)。与AngⅡ+sh-NC组比较,AngⅡ+sh-GRP78组细胞活性较低,迁移能力较低,CollagenⅠ、CollagenⅢ、α-SMA、GRP78、p-PERK、p-eIF2α和CHOP相对蛋白表达量较低(均P<0.05)。与AngⅡ+sh-GRP78组比较,AngⅡ+sh-GRP78+Tn组活性较强,迁移能力较强,CollagenⅠ、CollagenⅢ、α-SMA、GRP78、p-PERK、p-eIF2α和CHOP相对蛋白表达量较高(均P<0.05)。结论GRP78可以通过激活内质网应激来调节心肌梗死诱导的心肌纤维化中的成纤维细胞活性、迁移能力和纤维化进展。 展开更多
关键词 急性心肌梗死 心肌纤维化 葡萄糖调节蛋白78 内质网应激
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过表达葡萄糖调节蛋白78人乳腺癌细胞系HS578T、MDA-MB-231的构建
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作者 李艳敏 古丽拉莱·多力坤 +4 位作者 马西智 杨文月 单文豪 陈娜菲 周晓涛 《山东医药》 CAS 2024年第6期1-5,共5页
目的构建过表达内质网应激标志蛋白葡萄糖调节蛋白78(Glucose-regulated protein 78,GRP78)的人乳腺癌细胞系HS578T、MDA-MB-231,为探讨GRP78在乳腺癌发生发展中的作用机制提供基础。方法采用分子克隆技术构建表达GRP78的慢病毒载体pCDH... 目的构建过表达内质网应激标志蛋白葡萄糖调节蛋白78(Glucose-regulated protein 78,GRP78)的人乳腺癌细胞系HS578T、MDA-MB-231,为探讨GRP78在乳腺癌发生发展中的作用机制提供基础。方法采用分子克隆技术构建表达GRP78的慢病毒载体pCDH-CMV-GRP78-HA-EF1-Puro质粒,采用PCR法扩增质粒,后使用EcoRⅠ、NotⅠ限制性内切酶切割质粒,对酶切产物进行测序鉴定,成功构建pCDH-CMV-GRP78-HA-EF1-Puro质粒。取对数生长期293FT细胞系(克隆分离株),在培养皿中加入pCDH-CMV-GRP78-HA-EF1-Puro混合液,培养48 h收集含GRP78慢病毒颗粒的上清液,保存备用。另取对数生长期人乳腺癌细胞系HS578T及MDA-MB-231,置于含GRP78慢病毒颗粒上清液的培养基中培养,培养48 h时在培养基中加入1∶500稀释嘌呤霉素,筛选GRP78过表达的HS578T、MDA-MB-231细胞系。加入嘌呤霉素培养48 h采用Western Blotting法检测HS578T、MDA-MB-231细胞GRP78。采用激光共聚焦显微镜对HS578T、MDA-MB-231细胞中GRP78蛋白进行定位。结果构建表达GRP78的慢病毒载体pCDH-CMV-GRP78-HA-EF1-Puro质粒。GRP78过表达的HS578T、MDA-MB-231细胞系在78 kD左右可见明显GPR78蛋白电泳条带。HS578T细胞系中存在GRP78蛋白表达,主要分布在细胞质中。结论成功构建GRP78过表达的MDA-MB-231、HS578T细胞系,GRP78主要表达于细胞质中。 展开更多
关键词 葡萄糖调节蛋白78 乳腺癌 慢病毒载体
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