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Abstract ：
目的 探讨前导肽的融合对人锰超氧化物歧化酶(SOD2)结构以及抗顺铂(DDP)诱导的肾损伤效应.方法 通过结构预测和SOD比活力测定分析线粒体靶向序列(MTS)对SOD2构效的影响;建立昆明(KM)小鼠DDP损伤模型,以阿米福汀(AMFT)为阳性对照,测定小鼠肾功能、肾脏指数、肾脏抗氧化能力,同时观察肾脏表观及病理变化,以评估MTS-SOD2抗DDP诱导的肾损伤效应.结果 MTS前导肽对SOD2的二三级结构有一定影响,但也使MTS-SOD2蛋白的比活力提高.预给予中剂量MTS-SOD2(0.84 mg/kg)可以使损伤鼠肾脏丙二醛(MDA)水平显著降低,SOD活力和总抗氧化能力(T-AOC)显著增加,进而减少肾脏病理损伤,维持肾功能,整体效果与200 mg/kg AMFT相当甚至优于后者.结论 MTS前导肽既增强了SOD2的活性,又使其因具有跨膜功能而发挥优异的抗DDP诱导的肾损伤效应.
Keyword ：
前导肽 前导肽 肾损伤 肾损伤 锰超氧化物歧化酶 锰超氧化物歧化酶 顺铂 顺铂
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| GB/T 7714 | 潘剑茹 , 韩亚南 , 何夏琪 et al. 带前导肽的人锰超氧化物歧化酶抗顺铂诱导的肾损伤效应 [J]. | 肿瘤防治研究 , 2023 , 50 (7) : 675-680 . |
| MLA | 潘剑茹 et al. "带前导肽的人锰超氧化物歧化酶抗顺铂诱导的肾损伤效应" . | 肿瘤防治研究 50 . 7 (2023) : 675-680 . |
| APA | 潘剑茹 , 韩亚南 , 何夏琪 , 叶小强 , 何火聪 . 带前导肽的人锰超氧化物歧化酶抗顺铂诱导的肾损伤效应 . | 肿瘤防治研究 , 2023 , 50 (7) , 675-680 . |
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Annexin A2 (ANXA2) has been found to be involved in cancer proliferation, metastasis and prognosis; however, its exact role in nasopharyngeal carcinoma (NPC) radioresistance remains unknown. We found that ANXA2 expression was correlated with prognosis in NPC patients, and longer overall survival in NPC patients with low ANXA2 expression than those with high ANXA2 expression. ANXA2 knockdown increased the radiosensitivity in radioresistant NPC cells, and ANXA2 overexpression decreased the radiosensitivity in NPC cells. Knocking-down ANXA2 expression increased the irradiation-induced apoptosis of radioresistant NPC cells, and ANXA2 overexpression decreased the irradiation-induced apoptosis of NPC cells. ANXA2 knockdown induced G2/M phase arrest in NPC cells post-irradiation, and ANXA2 overexpression abrogated G2/M phase arrest in NPC cells post-irradiation. ANXA2 overexpression resulted in inhibition of the p38 MAPK-HSP27 pathway, while ANXA2 knockdown resulted in activation of the p38 MAPK-HSP27 pathway. In addition, ANXA2 knockdown increased the radiosensitivity of the xenografted tumors in nude mice. Our data demonstrate that knockdown of Annexin A2 enhanced radiosensitivity in NPC by increasing G2/M-phase arrest, apoptosis and activating the p38 MAPK-HSP27 pathway. ANXA2 may be a promising target used to overcome radioresistance in NPC.
Keyword ：
annexin A2 annexin A2 apoptosis apoptosis cell-cycle arrest cell-cycle arrest nasopharyngeal carcinoma nasopharyngeal carcinoma radioresistance radioresistance
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| GB/T 7714 | He, Huocong , Lin, Keyu , Zou, Changyan et al. Knockdown of Annexin A2 Enhances Radiosensitivity by Increasing G2/M-Phase Arrest, Apoptosis and Activating the p38 MAPK-HSP27 Pathway in Nasopharyngeal Carcinoma [J]. | FRONTIERS IN ONCOLOGY , 2022 , 12 . |
| MLA | He, Huocong et al. "Knockdown of Annexin A2 Enhances Radiosensitivity by Increasing G2/M-Phase Arrest, Apoptosis and Activating the p38 MAPK-HSP27 Pathway in Nasopharyngeal Carcinoma" . | FRONTIERS IN ONCOLOGY 12 (2022) . |
| APA | He, Huocong , Lin, Keyu , Zou, Changyan , Pan, Jianru , Fu, Wankai , Zhou, Yan et al. Knockdown of Annexin A2 Enhances Radiosensitivity by Increasing G2/M-Phase Arrest, Apoptosis and Activating the p38 MAPK-HSP27 Pathway in Nasopharyngeal Carcinoma . | FRONTIERS IN ONCOLOGY , 2022 , 12 . |
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Antioxidant enzymes fused with cell-penetrating peptides could enter cells and protect cells from irradiation damage. However, the unselective transmembrane ability of cell-penetrating peptide may also bring antioxidant enzymes into tumor cells, thus protecting tumor cells and consequently reducing the efficacy of radiotherapy. There are active matrix metalloproteinase (MMP)-2 or MMP-9 in most tumor cellular microenvironments. Therefore, a fusion protein containing an MMP-2/9 cleavable substrate peptide X, a cell-penetrating peptide R9, a glutathione S-transferase (GST), and a human Cu, Zn superoxide dismutase (SOD1), was designed and named GST-SOD1-X-R9. In the tumor microenvironment, GST-SOD1-X-R9 would lose its cell-penetrating peptide and could not enter tumor cells due to the cleavage of substrate X by active MMP-2/9, thereby achieving selected entering normal cells. The complete nucleotide sequence of SOD1-X-R9 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1. The pGEX4T-1-SOD1-X-R9 recombinant plasmid was obtained, and soluble expression of the fusion protein was achieved. GST-SOD1-X-R9 was purified by ammonium sulfate precipitation and GST affinity chromatography. The molecular weight of the fusion protein was approximately 47 kDa, consistent with the theoretical value. The SOD and GST activities were 2 954 U/mg and 328 U/mg, respectively. Stability test suggested that almost no change in either SOD activity or GST activity of GST-SOD1-X-R9 was observed under physiological conditions. The fusion protein could be partially digested by collagenase IV in solution. Subsequently, the effect of MMP-2/9 activity on transmembrane ability of the fusion protein was tested using 2D and 3D cultured HepG2 cells. Little extracellular MMP-2 activity of HepG2 cells was observed under 2D culture condition. While under the 3D culture model, the size and the MMP-2 activity of the HepG2 tumor spheroid increased daily. GST-SOD1-R9 proteins showed the same transmembrane efficiency in 2D cultured HepG2 cells, but the transmembrane efficiency of GST-SOD1-X-R9 in 3D cultured HepG2 spheres was reduced remarkably. This study provided a basis for further investigating the selectively protective effect of GST-SOD1-X-R9 against oxidative damage in normal cells. ©2022 Chin J Biotech, All rights reserved.
Keyword ：
antioxidant enzyme antioxidant enzyme cell permeable cell permeable expression in Escherichia coli expression in Escherichia coli MMP-2/9 sensitive MMP-2/9 sensitive purification purification stability stability
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| GB/T 7714 | He, H. , Lin, L. , Li, L. et al. Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征] [J]. | Chinese Journal of Biotechnology , 2022 , 38 (9) : 3515-3527 . |
| MLA | He, H. et al. "Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征]" . | Chinese Journal of Biotechnology 38 . 9 (2022) : 3515-3527 . |
| APA | He, H. , Lin, L. , Li, L. , Wu, L. , Lin, H. , Pan, J. . Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征] . | Chinese Journal of Biotechnology , 2022 , 38 (9) , 3515-3527 . |
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融合了跨膜肽的抗氧化酶可进入细胞,保护细胞免受放射损伤.然而跨膜肽的跨膜能力没有靶向性,其也可把抗氧化酶带入肿瘤细胞进而保护肿瘤细胞,降低放疗的效果.为此,根据多数肿瘤细胞微环境中存在活性基质金属蛋白酶(matrix metalloproteinase,MMP)-2或MMP-9的特点,在细胞跨膜肽R9与人铜、锌超氧化物歧化酶(superoxide dismutase 1,SOD1)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)之间融合 MMP-2/9的底物肽X,设计了融合蛋白GST-SOD1-X-R9.该蛋白在肿瘤微环境中可因MMP-2/9酶切底物肽X而失去跨膜肽,从而无法进入肿瘤细胞,进而只能进入正常细胞.全基因合成SOD1-X-R9序列,并将其插入原核表达载体pGEX-4T-1中,得到表达质粒,并实现了GST-SOD1-X-R9融合蛋白的可溶表达.GST-SOD1-X-R9经硫酸铵沉淀和GST亲和层析纯化,分子量约为47kDa,与理论值一致.纯化的融合蛋白的SOD活性和GST活性分别为2 954 U/mg和328 U/mgoGST-SOD1-X-R9的SOD活性或GST活性在生理条件下几乎没有变化.该融合蛋白在溶液中可被胶原酶Ⅳ部分水解.分别建立了2D和3D培养的HepG2细胞模型来检验肿瘤微环境中的MMP-2活力对该蛋白跨膜能力的影响.在2D培养模型中,HepG2的MMP-2活力极低,但在3D培养模型中,随着培养时间的增加,HepG2肿瘤球的体积变大,其胞外MMP-2活力也随之增强.GST-SOD1-X-R9在2D培养的HepG2细胞中具有和GST-SOD1-R9蛋白一样的跨膜效率,但在3D培养的HepG2细胞球中的跨膜能力大大降低.本研究为后续深入研究GST-SOD1-X-R9靶向防护正常细胞的氧化损伤效应奠定了基础.
Keyword ：
MMP-2/9敏感性 MMP-2/9敏感性 可跨膜 可跨膜 大肠杆菌表达体系 大肠杆菌表达体系 抗氧化酶 抗氧化酶 稳定性 稳定性 纯化 纯化
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| GB/T 7714 | 何火聪 , 林丽香 , 李玲玲 et al. 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征 [J]. | 生物工程学报 , 2022 , 38 (9) : 3515-3527 . |
| MLA | 何火聪 et al. "基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征" . | 生物工程学报 38 . 9 (2022) : 3515-3527 . |
| APA | 何火聪 , 林丽香 , 李玲玲 , 吴伦巧 , 林海英 , 潘剑茹 . 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征 . | 生物工程学报 , 2022 , 38 (9) , 3515-3527 . |
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Abstract ：
融合了跨膜肽的抗氧化酶可进入细胞，保护细胞免受放射损伤。然而跨膜肽的跨膜能力没有靶向性，其也可把抗氧化酶带入肿瘤细胞进而保护肿瘤细胞，降低放疗的效果。为此，根据多数肿瘤细胞微环境中存在活性基质金属蛋白酶(matrix metalloproteinase,MMP)-2或MMP-9的特点，在细胞跨膜肽R9与人铜、锌超氧化物歧化酶(superoxide dismutase 1,SOD1)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)之间融合MMP-2/9的底物肽X，设计了融合蛋白GST-SOD1-X-R9。该蛋白在肿瘤微环境中可因MMP-2/9酶切底物肽X而失去跨...
Keyword ：
MMP-2/9敏感性 MMP-2/9敏感性 可跨膜 可跨膜 大肠杆菌表达体系 大肠杆菌表达体系 抗氧化酶 抗氧化酶 稳定性 稳定性 纯化 纯化
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| GB/T 7714 | 何火聪 , 林丽香 , 李玲玲 et al. 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征（英文） [J]. | 生物工程学报 , 2022 , 38 (09) : 3515-3527 . |
| MLA | 何火聪 et al. "基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征（英文）" . | 生物工程学报 38 . 09 (2022) : 3515-3527 . |
| APA | 何火聪 , 林丽香 , 李玲玲 , 吴伦巧 , 林海英 , 潘剑茹 . 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征（英文） . | 生物工程学报 , 2022 , 38 (09) , 3515-3527 . |
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Background: Paraspeckle component 1 (PSPC1) is overexpressed in various cancer and correlated with poor survival in the patients. However, little is known about its expression and role in the progression of nasopharyngeal carcinomas (NPC). The purpose of this study is to examine PSPC1 expression in NPC and explore its role in clinical prognosis of radiation therapy. Methods: The association of PSPC1 expression with clinicopathological features of 109 NPC patients was examined using partial correlation analysis. Cancer tissues were obtained prior to clinical treatment. All cases were diagnosed and pathologically confirmed to be poorly differentiated or undifferentiated NPC without distant metastasis. The patients were then treated with radiation and followed-up. Survival analysis was performed. Results: Partial correlation analysis revealed that the PSPC1 expression in NPC was correlated with N classification, recurrence, prognosis and radiosensitivity in NPC patients, but not with the gender, age, pathohistological pattern, clinical stage, and T classification. The overexpression of PSPC1 was detected in 64 samples (58.72%). Kaplan-Meier survival analysis revealed that the overall survival (OS) was longer in NPC patients with PSPC1 low expression than that in those with PSPC1 high expression. Moreover, patients with the overexpression of PSPC1 had a low progression-free survival and distant metastasis-free survival rate, compared to those who had a low expression of PSPC1. Although not statistically significant, patients with high expression of PSPC1 had a lower locoregional recurrence-free survival rate than those with low expression, and the curves between the two groups was well separated. Conclusion: PSPC1 overexpression was associated with poor prognosis for NPC, which might be a novel useful biomarker to predict the response of NPC to radiation therapy and its clinical outcome.
Keyword ：
clinical prognosis clinical prognosis nasopharyngeal carcinoma nasopharyngeal carcinoma overexpression overexpression paraspeckle component 1 paraspeckle component 1 PSPC1 PSPC1 radiation radiation
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| GB/T 7714 | He, Huocong , Zhang, Lurong , Lin, Keyu et al. The Prognosis Value of PSPC1 Expression in Nasopharyngeal Cancer [J]. | CANCER MANAGEMENT AND RESEARCH , 2021 , 13 : 3281-3291 . |
| MLA | He, Huocong et al. "The Prognosis Value of PSPC1 Expression in Nasopharyngeal Cancer" . | CANCER MANAGEMENT AND RESEARCH 13 (2021) : 3281-3291 . |
| APA | He, Huocong , Zhang, Lurong , Lin, Keyu , Huang, Zhengrong , Zhou, Yan , Lin, Shaojun et al. The Prognosis Value of PSPC1 Expression in Nasopharyngeal Cancer . | CANCER MANAGEMENT AND RESEARCH , 2021 , 13 , 3281-3291 . |
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为了进一步研究当归(Angelica sinensis)生药中的蛋白质及其功能,通过80％硫酸铵沉淀、Sephadex G-50凝胶过滤层析、DEAE-Sepharose阴离子交换层析,首次从当归生药中纯化出两种分子量相近的蛋白(命名为ASPR-C-1和ASPR-C-2).ASPR-C-1和ASPR-C-2在SDS-PAGE上的分子量分别为17.33 kDa和17.18 kDa,在溶液中主要以单体形式存在,但会部分形成二聚体,二者均为糖蛋白,糖基含量分别为2.6％和8.2％.经基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-TOFTM)鉴定发现ASPR-C-1和ASPR-C-2均为病程相关(Pathogenesis-related 10,PR-10)家族蛋白,且具有核糖核酸酶活性,比活分别为73.60 U/mg和146.76 U/mg.两种蛋白的最适pH相近,均为5.6左右,但最适温度不同,ASPR-C-1的为50℃,ASPR-C-2的为60℃.二者虽然在60℃下都表现出最大的酶活力稳定性,但在更高的处理温度(80-100℃)下,ASPR-C-1迅速失活,最终仅余20％左右活力,ASPR-C-2则表现出良好的热稳定性,最终仍有80％左右活力.此外,Fe2+对二者的酶活性具有激活作用,而Ca2+、Mg2+、Zn2+、Mn2+、Ag+、Cu2+、EDTA、DTT和SDS则会不同程度地抑制二者的酶活性.研究结果为深入研究来自当归生药的PR-10蛋白的生物学功能奠定了基础.
Keyword ：
PR-10 PR-10 当归生药 当归生药 核糖核酸酶 核糖核酸酶 糖蛋白 糖蛋白 纯化 纯化
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| GB/T 7714 | 王香玲 , 李娴 , 何火聪 et al. 当归生药中两种PR-10蛋白亚型的纯化与表征 [J]. | 生物工程学报 , 2019 , 35 (1) : 159-168 . |
| MLA | 王香玲 et al. "当归生药中两种PR-10蛋白亚型的纯化与表征" . | 生物工程学报 35 . 1 (2019) : 159-168 . |
| APA | 王香玲 , 李娴 , 何火聪 , 李玲玲 , 吕迪 , 陈翠煌 et al. 当归生药中两种PR-10蛋白亚型的纯化与表征 . | 生物工程学报 , 2019 , 35 (1) , 159-168 . |
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Abstract ：
目的探讨葡萄糖调节蛋白GRP78蛋白表达与鼻咽癌临床特征及预后的关系。方法随机选取2012年2月至2012年7月福建省肿瘤医院初诊鼻咽癌患者的治疗前鼻咽肿瘤活检组织标本107例,中位年龄47岁,全部病例鼻咽部组织均经病理学确诊且无远处转移。免疫组织化学法检测GRP78蛋白在鼻咽癌肿瘤组织的表达水平。根据鼻咽癌患者的性别、年龄、病理类型、肿瘤分期、无远处转移生存时间和总生存时间等临床资料,分析GRP78蛋白表达与临床分期、肿瘤远处转移和总生存等的关系。结果全部鼻咽肿瘤活检组织标本中GRP78蛋白的阳性表达率为72.9%(78/107)。GRP78蛋白的表达水平与患者的性别、年龄、病理类型、临床分...
Keyword ：
GRP78 GRP78 临床分期 临床分期 临床意义 临床意义 低表达 低表达 活检组织 活检组织 高表达 高表达 鼻咽癌患者 鼻咽癌患者 鼻咽肿瘤 鼻咽肿瘤
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| GB/T 7714 | 何火聪 , 苏颖 , 林可焴 et al. GRP78在鼻咽癌组织的表达及其临床意义 [C] //2018年中国肿瘤标志物学术大会暨第十二届肿瘤标志物青年科学家论坛论文集 . 2018 . |
| MLA | 何火聪 et al. "GRP78在鼻咽癌组织的表达及其临床意义" 2018年中国肿瘤标志物学术大会暨第十二届肿瘤标志物青年科学家论坛论文集 . (2018) . |
| APA | 何火聪 , 苏颖 , 林可焴 , 陈超 , 邹长棪 , 潘剑茹 . GRP78在鼻咽癌组织的表达及其临床意义 2018年中国肿瘤标志物学术大会暨第十二届肿瘤标志物青年科学家论坛论文集 . (2018) . |
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本发明涉及天然中药蛋白领域，更具体地涉及一种当归蛋白在制备防护急性肝损伤药物中的应用，所述当归蛋白N端序列为GIQKTEVEAPSTVSA。所述当归蛋白对正常肝细胞L‑02具有显著的促增殖作用，在0.5 mg/mL的剂量下，可使L‑02细胞的增殖率达到163.14%。所述当归蛋白用于制备预防肝损伤药物，可显著提高损伤鼠肝脏CAT、GST和T‑AOC活力极显著地减轻肝脏MDA水平，显著改善损伤鼠肝脏的肝细胞坏死状况，进而显著降低损伤鼠血液ALT、AST活力及肝脏指数，使之基本恢复至正常组水平，从而显著减轻CCl4对小鼠肝脏的损伤。
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| GB/T 7714 | 潘剑茹 , 王香玲 . 一种当归蛋白在制备防护肝损伤药物中的应用 : CN201810421947.8[P]. | 2018/5/4 . |
| MLA | 潘剑茹 et al. "一种当归蛋白在制备防护肝损伤药物中的应用" : CN201810421947.8. | 2018/5/4 . |
| APA | 潘剑茹 , 王香玲 . 一种当归蛋白在制备防护肝损伤药物中的应用 : CN201810421947.8. | 2018/5/4 . |
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本发明涉及天然中药蛋白领域，更具体地涉及一种当归蛋白(ASPR)及其蛋白自组装纳米颗粒，该蛋白N端序列为GIQKTEVEAPSTVSA。ASPR蛋白在pH 8.0下，经过100℃加热15min，可自组装形成纳米颗粒。通过100 KDa超滤分离得到的蛋白自组装纳米颗粒ASPR‑NP平均粒径为154.9±27.2 nm。该纳米颗粒稳定性好，可跨膜进入细胞，到达线粒体。此外，ASPR可包埋小分子难溶药物阿魏酸(FA)，得到ASPR‑FA‑NP，其平均粒径为216.3±18.3 nm。ASPR‑FA‑NP的稳定性好，能够选择性促进正常肝细胞增殖同时抑制肝癌细胞增殖，且生物相容性好。
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| GB/T 7714 | 潘剑茹 , 李娴 . 一种当归蛋白自组装颗粒及应用 : CN201810421944.4[P]. | 2018/5/4 . |
| MLA | 潘剑茹 et al. "一种当归蛋白自组装颗粒及应用" : CN201810421944.4. | 2018/5/4 . |
| APA | 潘剑茹 , 李娴 . 一种当归蛋白自组装颗粒及应用 : CN201810421944.4. | 2018/5/4 . |
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