AMIGO2通过激活PI3K/AKT/mTOR信号通路促进鼻咽癌细胞的增殖

目的：探讨Ig 样结构域 2 黏附分子（adhesion molecule with Ig like domain 2,AMIGO2）在鼻咽癌（nasopharyngeal carcinoma,NPC）细胞增殖中的作用及其机制。方法: 选用2017年9月至11月福建省肿瘤医院收集的10例NPC组织和10例正常鼻 咽黏膜上皮组织标本,以及NPC细胞系CNE-1、CNE-2、SUNE-1、 6-10B、 C666-1和人永生化鼻咽黏膜上皮细胞株NP69, 用qPCR法 检测NPC组织和细胞中AMIGO2 mRNA的表达。构建慢病毒载体干扰AMIGO2表达, 用qPCR法验证其干扰效率; 用CCK-8 法、克隆形成及流式细胞术检测干扰AMIGO2表达对NPC细胞增殖、克隆形成和凋亡的影响, 用 Western blotting 检测干扰 AMIGO2 表达对 NPC 细胞增殖及 PI3K/AKT/mTOR 信号通路相关标志蛋白表达的影响。结果: AMIGO2在NPC组织和 CNE-2和SUNE-1细胞中高表达（均P<0.01）。慢病毒AMIGO2感染后,CNE-2和SUNE-1细胞的AMIGO2干扰效率均达50%以 上。干扰AMIGO2表达,显著降低CNE-2和SUNE-1细胞增殖及克隆形成能力（均P<0.01）、明显提高细胞的凋亡率（均P<0.01）; 降低 SUNE-1 细胞中PI3K、AKT和mTOR磷酸化蛋白的表达水平 （均P<0.01）、下调survivin 和 PCNA 蛋白的表达水平（均P<0.01）。 结论：AMIGO2通过激活PI3K/AKT/mTOR信号通路促进NPC细胞增殖并抑制其凋亡,提示AMIGO2可能是NPC治疗的潜 在靶点。     

Overproduction of Cyclin D1 is dependent on activated mTORC1 signal in nasopharyngeal carcinoma: Implication for therapy

Activated mTOR was implicated to play a role in the carcinogenesis of nasopharyngeal carcinoma (NPC). However, the mechanism of activated mTOR/Complex1(mTORC1) signaling pathway in NPC development has not been well established. In this study, we correlated the expression of mTORC1 signal molecules and Cyclin D1 in NPC. We also investigated the effect of blocking mTORC1 signal with rapamycin and mTOR siRNA on Cyclin D1 expression in CNE-2 cells, as well as cell apoptosis and viability. We found a positive association of mTORC1 signal molecules and Cyclin D1 in NPC. Also, we found blockage mTORC1 inhibited Cyclin D1 expression in CNE-2 cells and enhanced cell apoptosis. Our results suggested that mTORC1 signal pathway might be a potential target for NPC therapy.     

Inhibiting ERp29 expression enhances radiosensitivity in human nasopharyngeal carcinoma cell lines

ERp29 is an endoplasmic reticulum (ER) stress-inducible protein. It was found that ERp29 was highly expressed in several cancers and associated with resistance to oxidative and radiation stress, which may serve as a novel target for nasopharyngeal carcinoma (NPC) anticancer approach. In this study, we used immunohistochemistry to detect ERp29 expression in radioresistant and radiosensitive NPC tissues. As a result, ERp29 was up-regulated in radioresistant NPC tissues compared to radiosensitive NPC tissues. We also found that ERp29 knockdown attenuated radioresistance of NPC CNE-1 cells and ERp29 overexpression enhanced radioresistance of NPC CNE-2 cells. When exposed to radiation, ERp29 knockdown CNE-1 cells increased radiation-induced cell apoptosis and ERp29 overexpression CNE-2 cells reduced radiation-induced cell apoptosis. Further, we demonstrated that ERp29 up-regulated the expression of Hsp27. In conclusion, our study supports ERp29 could potentiate resistance to radiation in NPC cells, targeting of ERp29 is a rational strategy in treating radioresistant NPC.     

下调VEGF表达影响鼻咽癌细胞放射敏感度的机制

目的　应用RNA干扰技术抑制人鼻咽低分化鳞状上皮细胞癌细胞株CNE-2中血管内皮生长因 子（VEGF）表达,研究阻断VEGF基因表达对鼻咽癌细胞放射敏感度的影响及机制。方法 构建针对 VEGF的siRNA真核表达载体pU-VEGF-siRNA（CNE-2组）、1个阴性对照质粒（CNE-2/Neg-siRNA 组）、经脂质体转染至CNE-2细胞（CNE-2/VEGF-siRNA组）,用平板克隆形成实验检测在6MV-X线 0、2、4、6、8、10 Gy照射后克隆形成能力及通过单击多靶模型、线性二次模型拟合放射生物学参 数,流式细胞检测分析细胞周期和细胞凋亡的变化,用RT-PCR定量分析三组细胞中Cyclin D1、Cyclin E、P16和P53的mRNA表达。结果 经6MV-X线0、2、4、6、8、10 Gy照射后细胞存活率显著下降, CNE-2/VEGF-siRNA组细胞经D0和2 Gy照射后的存活分数明显低于CNE-2组、CNE-2/Neg-siRNA组; CNE-2/VEGF-siRNA组中G1/S期细胞周期阻滞更为明显。在Cyclin D1、Cyclin E、p16和p53基因中, Cyclin D1mRNA表达在放疗后6、12和24 h进行性升高,差异有统计学意义,而其他基因变化差异无统 计学意义,Western blot检测显示Cyclin D1蛋白表达在放疗后24 h明显升高。结论 下调VEGF表达可 增加鼻咽癌细胞的放射敏感度,其机制可能是通过Cyclin D1信号通路途径使细胞周期发生G1/S期阻滞。     

Control of stability of cyclin D1 by quinone reductase 2 in CWR22Rv1 prostate cancer cells

Aberrant expression of cyclin D1, frequently observed in human malignant disorders, has been linked to the control of G(1)→S cell cycle phase transition and development and progression in carcinogenesis. Cyclin D1 level changes are partially controlled by GSK-3β-dependent phosphorylation at threonine-286 (Thr286), which targets cyclin D1 for ubiquitination and proteolytic degradation. In our continuing studies on the mechanism of prostate cancer prevention by resveratrol, focusing on the role of its recently discovered target protein, quinone reductase 2 (NQO2), we generated NQO2 knockdown CWR22Rv1 using short hairpin RNA (shRNA)-mediated gene silencing approach. We found that, compared with cells expressing NQO2 (shRNA08), NQO2 knockdown cells (shRNA25) displayed slower proliferation and G(1) phase cell accumulation. Immunoblot analyses revealed a significant decrease in phosphorylation of retinoblastoma Rb and cyclin D1 in shRNA25 compared with shRNA08. Moreover, shRNA25 cells showed a 37% decrease in chymotrypsin-like proteasome activity. An increase in AKT activity was also observed in shRNA25, supported by a ～1.5-fold elevation in phosphorylation and ～50% reduction/deactivation of GSK-3α/β at Ser21/9, which were accompanied by a decrease in phosphorylation of cyclin D1 at T286. NQO2 knockdown cells also showed attenuation of resveratrol-induced downregulation of cyclin D1. Our results indicate a hitherto unreported role of NQO2 in the control of AKT/GSK-3β/cyclin D1 and highlight the involvement of NQO2 in degradation of cyclin D1, as part of mechanism of chemoprevention by resveratrol.     

Identification of ERp29 as a biomarker for predicting nasopharyngeal carcinoma response to radiotherapy

Radioresistance continues to be a major problem in the treatment of nasopharyngeal carcinoma (NPC). This study aimed to identify novel proteins associated with NPC radio-resistance. We used a mass spectrometry driven-proteomic strategy to identify novel proteins associated with NPC radio-resistance, and differential proteins were subsequently processed by bio-informatic analysis. As a result, twelve proteins were identified with aberrant expression in radioresistant (RR) NPC tissues compare to radiosensitive (RS) NPC tissues. Among these proteins, ERp29, Mn-SOD, HSP27 and GST ω1 were found to be significantly up-regulated in RR NPC tissues, and ERp29 was selected for further validation. Immunohistochemistry analysis confirmed that ERp29 was overexpressed in RR NPC tissues compared with RS NPC tissues. To prove the role of ERp29 in the induction of NPC radioresistance, ERp29 was down-regulated in the ERp29 enriched NPC cells CNE-1 and 6-10B by specific shRNA. Radiosensitivity was measured using cell proliferation assay and clonogenic survival assay, and cell apoptosis was measured using flow cytometric analysis. We found that ERp29 knockdown attenuated CNE-1 and 6-10B cell radioresistance and enhanced cell apoptosis. These results suggest that ERp29 associates with radioresistance in NPC, and ERp29 could be a potential biomarker for predicting NPC response to radiotherapy.     

Role of MIF/CXCL8/CXCR2 signaling in the growth of nasopharyngeal carcinoma tumor spheres

Macrophage migration inhibitory factor (MIF) and CXCL8 (also named IL-8) are strongly expressed in the tissues of nasopharyngeal carcinoma (NPC). However, their role in the growth of NPC has not been fully examined. This study aims to evaluate the functions of MIF and CXCL8 on the growth of NPC tumor spheres. The elevated expression of CXCL8 in tumor over normal tissues was confirmed in 37 pairs of biopsies from NPC patients. In the in vitro study, all the poorly differentiated NPC cell lines, including the EBV-positive C666-1, and the EBV-negative CNE-1, CNE-2, SUNE-1, HNE-1 and HONE-1 cells, were found to express CXCL8 and MIF. Therefore, the EBV-positive C666-1 cell was selected to examine for the role of MIF and CXCL8 in the growth of the NPC tumor spheres. Functional study showed that the growth of C666-1 tumor spheres, under the nutrient poor or growth factor supplemented culture conditions, could be inhibited by the CXCL8 specific peptide inhibitor. The growth of the tumor spheres could also be reduced by the CXCR2 specific inhibitor SB225002 or the PI3K/AKT inhibitor LY294002, indicating that the endogenously produced CXCL8 plays an autocrine role in the growth of the tumor spheres. Further mechanistic studies revealed that the gene expression of CXCL8 could be reduced by the MIF specific small interfering RNA (siRNA) or NF-κB inhibitor parthenolide, and the growth of tumor spheres was also reduced after MIF siRNA transfection. Taken together, the present study highlights the role of MIF/CXCL8/CXCR2 axis in the growth of NPC tumor spheres. Chemotherapeutic interference of this signaling pathway may help to control the growth of the NPC tumor.     

微小RNA hsa-miR-93靶向STAT3提高鼻咽癌对放疗敏感性的机制研究

目的 研究hsa-miR-93在鼻咽癌放疗抗拒过程中的作用及分子机制。方法 以人鼻咽癌细胞株CNE-1、CNE-2、CNE-2R、HONE1、C666.1以及鼻咽上皮永生细胞NP460为研究对象,采用实时荧光定量PCR(qRT-PCR)法检测各组细胞经放疗后hsa-miR-93的表达;采用Western blot法检测各组细胞经放疗后STAT3蛋白的表达;通过miRWalk软件和RNAHybrid软件预测hsa-miR-93和STAT3的结合作用及结合位点;通过双荧光素酶报告基因法检测hsa-miR-93对STAT3靶向结合情况;瞬时转染小分子模拟物或抑制剂调控hsa-miR-93的表达后,采用qRT-PCR和Western blot法检测STAT3的表达;转染siSTAT3沉默STAT3的表达后,通过细胞集落形成实验检测鼻咽癌细胞经放疗后的集落形成能力。结果 与CNE-1、CNE-2细胞(相对放疗敏感)相比,hsa-miR-93在HONE1、CNE-2R细胞(相对放疗抗拒)中表达降低(P＜0.001),且各组鼻咽癌细胞经放疗后hsa-miR-93的表达呈剂量及时间依赖性降低(P＜0.05);与CNE-1、CNE-2细胞相比,STAT3在HONE1、CNE-2R细胞中表达升高,且各组鼻咽癌细胞经放疗后表达呈剂量及时间依赖性升高;经预测,hsa-miR-93可直接靶向结合STAT3。过表达hsa-miR-93可以在mRNA以及蛋白水平上显著抑制STAT3的表达(P＜0.05),而抑制hsa-miR-93的表达可以显著上调STAT3的表达(P＜0.05);过表达hsa-miR-93或沉默STAT3均使鼻咽癌细胞的集落形成显著减少(P＜0.001);抑制hsa-miR-93减弱了鼻咽癌对放疗的敏感性,而同时抑制hsa-miR-93和STAT3后,可提高鼻咽癌对放疗的敏感性。结论 hsa-miR-93可能通过靶向STAT3提高鼻咽癌对放疗的敏感性。     

PD-L1 Induces Epithelial–Mesenchymal Transition in Nasopharyngeal Carcinoma Cells Through Activation of the PI3K/AKT Pathway

Nasopharyngeal cancer (NPC) is a malignant epithelial carcinoma of the head and neck. Cancer therapy targeting programmed cell death protein-1 (PD-1) or programmed death ligand-1 (PD-L1) is revolutionary. However, the tumorigenic mechanism of PD-L1 is not yet clear in NPC. Here we demonstrated an oncogenic role of PD-L1 via activating PI3K/AKT in NPC cells. PD-L1 overexpression was frequently detected in NPC biopsies and cell lines by qRT-PCR. PD-L1 overexpression and knockdown demonstrated that PD-L1 promoted NPC cell invasion and metastasis in vitro and in vivo. Mechanistically, PD-L1 prominently activated the epithelial–mesenchymal transition (EMT) process in a PI3K/AKT-dependent manner. Taken together, we found that PD-L1 overexpression confers NPC cell malignancy and aggressiveness via activating the downstream PI3K/AKT signaling. Thus, these results provide a basis for diagnosis and treatment of NPC.     

PAI-1在鼻咽癌细胞系(CNE-2Z)及其克隆株(CNE-2Z-H5,CNE-2Z-H5-9)中表达和意义

目的:探讨尿激酶激活剂抑制剂-1(Plasminogenactivatorinhibitor-1,PAI-1)在不同淋巴道转移能力的鼻咽癌细胞系(CNE-2Z)及其克隆株(CNE-2Z-H5,CNE-2Z-H5-9)中的表达和意义。方法:应用逆转录聚合酶反应(RT-PCR)方法检测PAI-1在基因转录水平的表达;使用异质粘附、侵袭抑制实验观察PAI-1在鼻咽癌细胞侵袭和转移过程中的作用。结果:PAI-1在CNE-2Z,CNE-2Z-H5中均有转录,但无显著性差异;PAI-1在CNE-2Z-H5-9中无转录。抗PAI-1抗体对CNE-2Z-H5-9异质粘附、侵袭无影响。结论:PAI-1可能抑制鼻咽癌的侵袭和转移。     

CircLASP1 silence strengthens the therapeutic effects of MK-2206 on nasopharyngeal cancer through upregulating miR-625

Therapeutic effects of MK-2206 are largely limited due to the complexity of the pathogenesis of nasopharyngeal cancer (NPC). Here, we aimed to investigate whether and how circLASP1 is involved in the therapeutic effects of MK-2206 on NPC. We showed circLASP1 was increased while miR-625 was decreased in NPC tissues and cell lines. CircLASP1 silence strengthened the therapeutic effects of MK-2206 via suppressing NPC cell proliferation and inducing autophagy and apoptosis in vitro. In mechanism analyses, we found that circLASP1 indirectly released AKT by directly binding to miR-625 in NPC cells, and miR-625 acted as a tumor suppressor in NPC and activated cell autophagy through inhibiting the AKT/mTOR pathway. Most importantly, knockdown of circLASP1 was revealed to enhance the therapeutic effects of MK-2206 on NPC in vivo. Our results suggest that the circLASP1/miR-625 axis is involved the therapeutic effects of MK-2206 on NPC by regulating autophagy, proliferation, and apoptosis through the AKT/mTOR pathway. miR-625 is involved in NPC tumorigenesis.     

Squalene epoxidase‐induced cholesteryl ester accumulation promotes nasopharyngeal carcinoma development by activating PI3K/AKT signaling

Nasopharyngeal carcinoma (NPC) is a common malignant tumor and a major cause of mortality and morbidity in southern China. However, the mechanism is still elusive. Here, we focused on studying the role of squalene epoxidase (SQLE), a key enzyme of cholesterol biosynthesis, in the progression of NPC. Clinical study revealed that SQLE expression was significantly upregulated in NPC tissues compared to normal tissues from mRNA level and patients with high expression of SQLE showed a poor prognosis. In vitro experiments showed that SQLE overexpression led to a significant proliferation of cells whereas SQLE knockdown showed an opposite result. In vivo studies also showed that SQLE promoted tumor growth in nude mice. Further study revealed that SQLE promoted NPC proliferation by cholesteryl ester accumulation instead of cholesterol. Mechanism studies indicated that cholesteryl ester promoted NPC cell proliferation by activating the PI3K/AKT pathway and inhibition of this pathway in SQLE‐overexpressed or cholesteryl ester‐treated cells resulted in a significant reduction of NPC cell proliferation. These results indicate that the oncogenic effect of SQLE in NPC mainly resulted from cholesteryl ester accumulation and PI3K/AKT is a promising target for NPC with SQLE overexpression.     

二苯并呫吨类化合物CY-B12体外抗肿瘤细胞增殖的作用及其机制的初步研究

背景与目的:CY-B12是最近合成的新型二苯并呫吨类化合物,本研究探讨CY-B12的体外抗肿瘤细胞增殖作用及其机制。方法:MTT法测定CY-B12体外抑制胃癌细胞MGC803、鼻咽癌细胞CNE-2、口腔上皮癌细胞KB-3-1及肺癌细胞Glc82等细胞株增殖的作用,流式细胞仪检测CY-B12作用后细胞周期的改变及细胞凋亡,Hoechst33258染色后荧光显微镜观察细胞凋亡形态,Westernblot测定细胞周期相关蛋白Cdc25C、CyclinB1、Cdc2的改变,单细胞凝胶电泳检测CY-B12对细胞DNA的损伤作用。结果:CY-B12对MGC803、CNE-2、KB-3-1及GLC82等细胞株的增殖都有明显的抑制作用,IC50值分别为7.51、9.58、8.84和15.99μmol/L。CY-B12可诱导CNE-2细胞周期阻滞于G2/M期继而发生细胞凋亡,可观察到细胞染色质凝集、凋亡小体产生及凋亡峰出现。CY-B12作用24h后,细胞周期相关蛋白Cdc25C表达水平随CY-B12浓度升高而下调;CyclinB1蛋白在CY-B12浓度低时升高,CY-B12浓度高时表达下降;Cdc2蛋白变化趋势与CyclinB1相同。单细胞凝胶电泳证明CY-B12诱导了DNA损伤,并存在浓度-效应依赖关系。结论:CY-B12有较强的体外抗增殖作用,它可能通过断裂细胞DNA、下调周期相关蛋白Cdc25C、诱导细胞发生周期阻滞和凋亡而发挥其作用。     

细胞角蛋白13通过PTEN抑制PI3K/AKT/mTOR通路增强鼻咽癌HNE1细胞放疗敏感性

目的:探讨细胞角蛋白13(cytokeratin13,CK13)对鼻咽癌HNE1细胞放疗敏感性的影响及其作用机制.方法:将HNE1细胞分为对照组、anti-CK13#a组及anti-CK13#b组(敲减CK13)、对照组+西罗莫司处理组(100nmol/L的西罗莫司处理1 h)、anti-CK13#a+西罗莫司处理组(...     

miR-107靶向FoxM1对鼻咽癌细胞生长和侵袭及迁移影响

目的microRNA与鼻咽癌(nasopharyngeal carcinoma,NPC)发生发展密切相关,但miR-107在鼻咽癌中的作用机制仍不清楚。本研究检测miR-107在鼻咽癌组织中的表达水平,并初步探讨其对鼻咽癌细胞增殖、迁移和侵袭影响的分子机制。方法收集2013-02-02-2013-12-24于佛山市第一人民医院经病理活检确诊的86例NPC组织和42例慢性鼻咽炎组织。采用原位杂交技术检测组织中miR-107的表达。分析miR-107表达与鼻咽癌患者临床特征及5年生存率的关系。双荧光素酶实验检测miR-107和人叉头框蛋白M1(forkhead box M1,FoxM1)在鼻咽癌细胞中的相互作用。在鼻咽癌细胞系CNE-2中转染miR-107 mimics和mimics NC,并设立mock对照组,采用实时荧光定量PCR法检测FoxM1 mRNA的表达量,蛋白质印迹实验检测FoxM1蛋白的表达量。然后通过CCK8、划痕和Transwell小室法检测鼻咽癌细胞的增殖、迁移和侵袭能力。结果miR-107在鼻咽癌组织中的阳性表达率(15.12%,13/86)低于慢性鼻咽炎组织(83.33%,35/42),差异有统计学意义,χ^2=96.37,P<0.001;与T分期(χ^2=24.816,P<0.001)和N分期(χ^2=29.368,P<0.001)有关联,且鼻咽癌miR-107阳性患者的5年生存率高于miR-107阴性组,χ^2=6.380,P=0.043。miR-107靶向调控FoxM1的3′UTR,降低了荧光素酶的表达活性,F=363.600,P<0.001。miR-107 mimics组FoxM1mRNA(F=267.600,P<0.001)和蛋白表达水平(F=101.300,P<0.001)低于mimics NC和mock组。与mimics NC和mock组相比,miR-107 mimics组细胞的增殖(F=1192.000,P<0.001)和迁移、侵袭(F=210.600,P<0.001)能力均降低。结论miR-107在鼻咽癌患者中低表达,通过靶向抑制FoxM1的表达而抑制鼻咽癌细胞的增殖、迁移和侵袭能力。     

AKT3 is a key regulator of head and neck squamous cell carcinoma

The phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway plays a vital role in cell proliferation, apoptosis, metabolism, and angiogenesis in various human cancers, including head and neck squamous cell carcinoma (HNSCC). In the present study, we aimed to clarify the role of AKT, which is a major downstream effector of the PI3K-AKT-mTOR pathway, in HNSCC. We first investigated the mRNA expression of AKT isoforms using RNA-sequencing data from The Cancer Genome Atlas database. We observed a specific elevation of AKT3 expression in HNSCC tissues when compared with that in normal tissues. Furthermore, AKT3 expression correlated with genes related to the immunosuppressive microenvironment more than the other AKT isoforms and PIK3CA. Accordingly, we focused on AKT3 and performed a knockdown approach using an HNSCC cell line. AKT3 knockdown cells exhibited impaired proliferation, a shift in the cell cycle from G2/M to G1/G0 phase, an increase in apoptotic cells, and downregulation of gene expression related to immunosuppression, as well as the knockdown of its upstream regulator PIK3CA. We also performed immunohistochemistry for both AKT3 and PIK3CA using surgical specimens from 72 patients with HNSCC. AKT3 expression in tumor cells correlated with immune cell infiltration and unfavorable prognosis when compared with PIK3CA. These findings suggested that AKT3 expression is a potential biomarker for predicting the immunoreactivity and prognosis of HNSCC. Furthermore, the isoform-specific inhibition of AKT3 could be developed as a novel cancer therapy that efficiently suppresses the PI3K-AKT-mTOR pathway.     

非小细胞肺癌组织中AKT2、Cyclin D1、MMP-9表达及其与临床病理因素的关系

背景与目的：AKT2是一种丝氨酸／苏氨酸激酶,已被确定为癌基因,在多种肿瘤组织中都存存AKT2的异常表达和活化,AKT2与肿瘤的增殖和侵袭转移密切相关。本研究旨在探讨AKT2、Cyclin D1、MMP-9存非小细胞肺癌（nonsmall cell lung cancer,NSCLC）中的表达及与临床病理因素的关系。方法：应用免疫组化的方法检测68例NSCLC组织、38例相应的癌旁组织和14例非肿瘤性肺组织标本中AKT2蛋白、Cyclin D1、MMP-9的表达,采用X^2检验分析临床病理因素与上述指标的相关性。结果：AKT2、Cyclin D1、MMP-9在NSCLC组织中阳性率分别为91．2％、76．5％、72．1％,显著高于癌旁及非肿瘤性肺组织中阳性率（3．8％、0％、13．5％）（P〈0．05）。AKT2的表达与患者年龄、性别、肿瘤类型及分化程度、TNM分期无关（P〉0．05）,与淋巴结转移有关（P〈0．05）。Cyclin D1、MMP-9表达与淋巴结转移、鳞癌的分化程度有关（P〈0．05）,MMP-9还与TNM分期有关（P〈0．05）。AKT2的表达与Cyclin D1、MMP-9呈正相关。结论：AKT2、Cyclin D1、MMP-9均与肺癌的发展有关,Cyclin D1、MMP-9的高表达可能与AKT2的调节有关。     

鼻咽癌细胞系中核迁移蛋白的表达及作用

背景与目的:核迁移是真核生物生长发育和细胞功能所必需的,核迁移蛋白(nucleardistributionC,NUDC)在核迁移中起着重要作用。本研究旨在检测NUDC在鼻咽癌细胞系CNE-2、HNE-2中的表达情况,初步探讨其抗体对癌细胞活性的影响。方法:在大肠杆菌中表达GST-NUDC重组蛋白并制备其多克隆抗体,间接免疫荧光染色法明确NUDC蛋白在鼻咽癌细胞中的亚细胞定位,Westernblot确定NUDC蛋白在鼻咽癌细胞系中表达情况,MTT法检测NUDC抗体对鼻咽癌细胞生长的影响。结果:鼻咽非癌组织及鼻咽癌细胞中均有NUDC蛋白表达,且其在鼻咽癌细胞中的表达量明显高于鼻咽非癌组织。NUDC蛋白在鼻咽癌细胞中主要位于细胞质核区附近,细胞核也可见少量表达。NUDC蛋白抗体可抑制鼻咽癌细胞生长,且存在着剂量-反应关系。结论:NUDC蛋白在鼻咽癌细胞系中高度表达,且NUDC蛋白抗体在体外可抑制鼻咽癌细胞的生长。