Nd-YAG激光汽化结合光动力学治疗对鼻咽癌患者SIL-2R、IL-2水平及NK活性的影响

应用双抗体夹心ＥＬＩＳＡ法、３Ｈ－ＴｄＲ掺入法和１２５Ｉ－ＵｄＲ释放法对２４例鼻咽癌（ＮＰＣ）患者ＹＡＧ激光汽化结合光动力学治疗前后外周血可溶性ＩＬ－２受体（ＳＩＬ－２Ｒ）、白细胞介素２（ＩＬ－２）水平和自然杀伤细胞（ＮＫ）活性分别进行测定。结果表明鼻咽癌患者在接受激光光动力学治疗后ＳＩＬ－２Ｒ水平呈明显下降（Ｐ＜０．０００５），而ＩＬ－２水平和ＮＫ活性显著上升（Ｐ＜０．０００５）。结果提示激光光动力学治疗对恢复鼻咽癌患者免疫功能起促进作用。     

Preliminary results of photodynamic therapy for recurrent nasopharyngeal carcinoma

Photodynamic therapy (PDT) is a promising new modality in the treatment of cancer. In Hong Kong where nasopharyngeal carcinoma (NPC) is endemic, radiotherapy has been the primary treatment of choice. For recurrent disease after radiotherapy, there is no effective treatment. This latter report summarizes our initial experience in using PDT for these patients. Twelve patients (three females and nine males) with ages ranging from 33 to 65 years were treated with an infusion of hematoporphyrin derivative (5 mg/kg) 48–72 h before exposure to 200 J/cm² light (wavelength, 630 nm) delivered from a gold vapor laser. All 12 patients showed a dramatic response as judged by computed tomography or magnetic resonance imaging at 6 months post-PDT. Of the eight patients in whom cure was aimed for, three remained disease-free at 9–12 months after a single treatment. Three of the remaining four patients achieved useful palliation. Skin hypersensitivity occurred in two patients and was the only significant complication encountered. This experience indicates that PDT can be an encouraging palliative or definitive management for recurrent superficial NPC.     

bcl-xs增加鼻咽癌细胞对喜树碱诱导凋亡敏感性的实验研究

目的 :研究 bcl- xs 基因对喜树碱 (CPT)诱导鼻咽癌 CNE- 2 Z细胞凋亡的影响。方法 :利用脂质体L ipofect Amine将含有人 bcl- xs基因的重组真核表达质粒 pc DNA3xs导入人鼻咽癌低分化上皮细胞株 CNE- 2 Z,G418筛选培养后 ,经 Western blot检测 bcl- xs的表达。以不含有 bcl- xs基因的 pc DNA3质粒转染 CNE- 2 Z作为对照细胞。喜树碱处理两种细胞一定时间后 ,通过激光流式细胞仪检测凋亡细胞百分比。结果 :Western blot检测证实获得稳定表达 bcl-xs的细胞 ,经过 0 .5μmol/L ,1.0μmol/L CPT处理 2 4小时后 ,激光流式细胞仪检测的凋亡峰值均高于对照细胞。结论 :bcl- xs能显著增加鼻咽癌 CNE- 2 Z细胞对喜树碱诱导凋亡的敏感性     

COX-2蛋白在鼻咽癌中的表达及其临床意义

目的 探讨COX-2(cyclooxygenase-2)蛋白在鼻咽癌组织中的表达及其临床意义。方法 分别应用免疫组化方法（IHC）和蛋白印迹方法（Western Blot）检测47例鼻咽癌组织（其中用IHC33例,Western Blot14例）及24例鼻咽部炎症组织中COX-2蛋白的表达。结果 COX-2蛋白在鼻咽癌组织的阳性表达率为81.8%（27/33）,在鼻咽炎组织的阳性表达率为25%（3/12）,两者表达差异有统计学意义（P=0.001）。Western Blot结果显示,在两种组织中COX-2蛋白表达的相对值分别为1.258±0.034和0.487±0.076,在鼻咽癌组表达显著增高（P=0.000）。结论 COX-2蛋白的高度表达在鼻咽癌的发生、发展中起重要作用,选择性COX-2抑制剂有望成为鼻咽癌化疗的重要药物。     

靶向c-Met嵌合抗原受体T细胞的制备及其对鼻咽癌细胞作用的研究

目的 构建靶向c-Met的嵌合抗原受体(CAR)逆转录病毒载体,制备靶向c-Met CAR-T,观察其对c-Met阳性鼻咽癌细胞的杀伤作用。 方法 利用基因工程技术构建抗c-Met scFv,并将其重组到含有CD28,CD137,CD3ζ等的逆转录病毒载体中,形成c-Met CAR逆转录病毒载体,测序确认。以该病毒载体感染T淋巴细胞,通过Western blotting检测c-MetCAR在T淋巴细胞中的表达。CCK-8检测c-Met CAR-T对鼻咽癌细胞的作用;通过ELISA检测c-Met CAR-T作用后IFN-γ、IL-2的变化。 结果 测序结果显示c-Met CAR病毒载体序列正确;Western blotting可检测到CD3ζ的表达;CCK-8结果显示,c-Met CAR-T明显抑制c-Met阳性的鼻咽癌细胞的增殖(P<0.05);ELISA结果显示,c-Met CAR-T作用后分泌IFN-γ、IL-2明显增加(P<0.01)。 结论 成功制备了靶向c-Met的嵌合抗原受体逆转录病毒载体。该嵌合抗原受体能在T细胞中表达,能有效抑制c-Met阳性鼻咽癌细胞增殖,增加IFN-γ、IL-2的分泌。     

鼻咽癌放/化疗后吞咽障碍的临床评估

目的 遴选出安全、有效、便捷的鼻咽癌放/化疗后吞咽障碍的评估方法。方法 选取在邵阳市中心医院就诊的鼻咽癌患者37例,应用吞咽X线荧光透视检查（VFSS）、进食评估问卷调查量表（EAT-10）、安德森吞咽障碍量表（MDADI）、反复唾液吞咽试验（RSST）和洼田饮水试验（WST）对入组患者进行吞咽功能的评估,筛选适合鼻咽癌放/化疗后吞咽障碍评估的方法。结果 以VFSS为金标准,进食评估EAT-10的灵敏度为83.33%,MDADI灵敏度为72.22%,RSST特异度为84.21%,但这些方法与金标准的Kappa值都小于0.2,其一致性较低。WST特异度为78.95%,Kappa值大于0.2,具有相对较好的一致性。结论 WST与VFSS的一致性相对较好,可以作为鼻咽癌放/化疗后吞咽障碍的筛查工具,其他方法也可辅助评估。     

Identification of a plasminogen activator derived from nasopharyngeal carcinoma

Summary The activity of plasminogen activator (PA) in tissue extracts from nasopharyngeal carcinoma (NPC) was determined by means of the fibrin plate method. Development of PA activity was observed in 16 out of 25 cases investigated. Furthermore, using tissue extract of NPC with PA activity, characterization and identification of the activator were carried out by means of electrophoretic analysis, fibrin zymography and immunological analysis. The molecular weight of this PA was found to be 38,000 daltons. Additionally, a urokinase type of plasminogen activator was contained in the tissue extracts.     

Current understanding and management of nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) is a kind of rare head and neck cancer in Japan. However, NPC has some unique features. It is one of the most popular cancers in southern China, Southeast Asia, the Arctic, and the middle East/north Africa. This distinctive racial, ethnical, and geographic predisposition to NPC implies that both genetic susceptibility and environmental factors contribute to the development of this tumor. NPC is an Epstein-Barr virus - associated tumor. Consistent elevation of EBV antibody titers is a well-established risk factor of development of NPC. Not only pathophysiological relationship, but also molecular mechanism of EBV-mediated carcinogenesis has been enthusiastically investigated. LMP1, an EBV primary oncogene, upregulates each step of metastasis, and contribute to highly metastatic feature of NPC. A tumor suppressor gene p53 is mostly intact and overexpressed in NPC whereas expression of p16, a cyclin-dependent kinase inhibitory protein, is downregulated in 2/3 of NPC. Intention modulated radiotherapy (IMRT) is now getting prevalent for the treatment of NPC because of complicated structure and location of nasopharynx. A good therapeutic result can be achieved by distributing a high dose to the tumor while keeping down normal tissue complications by reducing radiation dose to normal tissues. Chemotherapy is important to control distant metastasis of chemoradiosensitive NPC, and thus, should play an important role. However, most effective combination of anti-tumor drugs, protocol of chemoradiotherapy has not well-established. Finally, molecular targeting therapy, including targeting EBV gene product, has been developing and on the way to the clinical use.     

SHCBP1高表达促进鼻咽癌细胞上皮间质转化过程的分析

目的 探究SHC SH2结构域结合蛋白1(SHCBP1)在鼻咽癌组织及鼻咽癌细胞系中的表达情况,分析其对鼻咽癌细胞系的增殖、侵袭能力及上皮间质转化(EMT)过程的影响。方法 利用TCGA数据库分析SHCBP1在头颈鳞状细胞癌(HNSC)中的表达情况,分析SHCBP1表达程度与肿瘤免疫细胞浸润程度的关系。同时收集2019年9月—2021年9月于贵州医科大学附属医院及贵州医科大学附属肿瘤医院经病理确诊的31例鼻咽癌初诊患者临床组织样本,所有患者就诊前均未接受任何放化疗,并收集同期就诊的31例疑似鼻咽癌但病检示慢性鼻咽炎组织作为对照组。利用逆转录实时荧光定量PCR(RT-qPCR)检测临床样本及鼻咽癌细胞系5-8F中SHCBP1的表达情况。细胞实验以鼻咽癌细胞系5-8F、正常鼻咽部永生化上皮细胞系NP69作为研究对象。采用利用慢病毒载体shRNA-SHCBP1干扰技术沉默5-8F细胞中SHCBP1表达,并分为NC组(空载慢病毒LV-Zs-Green-PURO-NC感染5-8F)、SHCBP1-shRNA组(LV-SHCBP1-shRNA-Zs-Green-PURO沉默5-8F细胞SHCBP1表达)。CCK-8法、克隆形成实验检测SHCBP1对鼻咽癌细胞增殖的影响;划痕实验、Transwell迁移、侵袭实验检测SHCBP1对鼻咽癌细胞的转移、侵袭能力的影响。Western blot检测NC组、SHCBP1-shRNA组中E-cadherin、N-cadherin、基质金属蛋白9(MMP9)、Vimentin的表达情况。结果 在HNSC中SHCBP1高表达,表达量与Th2细胞浸润程度相关。与慢性鼻咽炎组织相比,鼻咽癌组织中SHCBP1明显高表达,差异具有统计学意义(P<0.05)。与NC组相比,SHCBP1-shRNA组鼻咽癌细胞增殖、迁移、侵袭能力均减弱,差异具有统计学意义(P<0.05)。SHCBP1-shRNA组E-cadherin表达较NC组升高,N-cadherin、MMP9、Vimentin表达较NC组降低,差异具有统计学意义(P<0.05)。结论 SHCBP1在鼻咽癌中高表达,并可促进鼻咽癌细胞增殖、侵袭、迁移及激活EMT过程。     

Induction of apoptosis by high-dose gold nanoparticles in nasopharyngeal carcinoma cells

Objective

Nasopharyngeal carcinoma (NPC) is a rare malignancy in most parts of the world, but is a common cancer in southern Asia. Local recurrent disease and distant metastasis of NPC are still the unsolved problems. Recently, gold nanoparticles (AuNPs) have been developed as potential in vivo diagnostic and therapeutic agents. However, their role on nasopharyngeal cancer remains unknown. The object of this study is to investigate if AuNPs can be used as a new therapeutic agent for NPC by evaluating their anti-tumor effect in vitro.

Methods

The AuNPs were prepared by the reduction of chloroauric acid to neutral gold. Their size distribution and microstructures were characterized by transmission electron microscopy (TEM). To evaluate their cytotoxic effect, NPC cell line TW01 and Human Nasal Epithelial Cells (HNEpC) were cultured in various concentrations of AuNPs for 3 days. Cell viability was evaluated by Trypan Blue viability assay while morphologic findings were observed via light microscopy. Terminal deoxynucleotidyltransferase-mediated dUPT nick end labeling (TUNEL) assay was used to detect apoptosis.

Results

AuNPs prepared in this study had an average diameter of 20.5 nm and they were observed under light microscopy as dark material aggregated in the cells after treatment. Contrary to the HNEpC, the AuNPs reduced cell viability of NPC cell in a concentration-dependant manner by Trypan Blue assay, especially at high concentration. Besides, cell apoptosis was demonstrated by positive TUNEL assay.

Conclusions

The AuNP possesses specific imaging properties and is cytotoxic to NPC cells at high concentrations.