基于表面增强拉曼光谱(SERS)技术对于人体细胞组织与血液的检测和研究, SERS光谱技术能够发现正常组织与病变组织的差异性, 为医学临床上实现癌症的早期诊断提供了科学依据。由于鼻咽癌不具有明显的病变特征、病灶的位置难以通过常规医学手段检测。因此利用SERS光谱技术, 应用于鼻咽癌细胞组织与血液的研究, 可提高鼻咽癌患者的生存率。对鼻咽癌细胞组织与血液SERS光谱分析和诊断的探索研究, 有助于SERS光谱技术发展成为一种在生物医学领域中的分析检测手段, 在医学临床诊断上具有潜在的应用前景。

Latent membrane protein 1 (LMP1) is known as an oncoprotein in nasopharyngeal carcinoma (NPC) cells, which is considered to have a strong association with growth, invasion and metastasis of NPC cells through lipid rafts. Methyl-β-cyclodextrin (MβCD) can disrupt lipid rafts through cholesterol depletion. In this study, we revealed that MβCD induced apoptosis in two kinds of NPC cells including CNE1 cells, a LMP1-negative nasopharyngeal squamous carcinoma cell line, and CNE1-LMP1 cells, a LMP1-positive nasopharyngeal squamous carcinoma cell line. Furthermore, the impact of MβCD on LMP1 was investigated by fluorescence resonance energy transfer (FRET) method in NPC cells. Synchronized tempo-spatial and spectral detection of LMP1/LMP1 interaction were performed using fluorescence microscope and spectrograph. FRET efficiency indicated that LMP1/LMP1 interaction gradually enhanced after MβCD treatment. MTT assays showed that MβCD caused strong cytotoxicity in CNE1 cells, but caused relatively weaker cytotoxicity in CNE1-LMP1 cells, which indicated that LMP1 may regulate sensitivity of NPC cells to MβCD. Then, detection of cleaved caspase-3 in two kinds of NPC cells indicated that LMP1 may inhibit activation of caspase-3. These results strongly suggested that MβCD can induce apoptosis of NPC cells, but enhancing of LMP1/LMP1 interaction may likely resist apoptosis through inhibiting cleavage of caspase-3.

表面增强拉曼光谱（SERS）技术是一种快速、 有效、 无损的光学诊断方法， 能提供生物分子成分和结构的指纹特征。 人的唾液包含多种能反应人体健康状况的生物标志物。 采用纳米银溶胶为增强基底的SERS技术， 从46位鼻咽癌患者、 30位鼻炎患者和48位健康人的唾液样本中采集了208条高质量的SERS。 研究结果表明鼻咽癌患者、 鼻炎患者和健康人的唾液SERS之间存在明显差异。 利用主成分分析结合线性判别分析(PCA-LDA)对采集的唾液SERS进行诊断研究， 获得灵敏度分别为89.5%， 93.0%和100%， 特异性分别为94.6%， 96.7%和97.7%。 研究结果表明： 唾液SERS技术结合PCA-LDA分析方法可能快速、 有效区分鼻咽癌、 鼻炎患者与正常人的唾液样本， 具有鼻咽癌无损检测和临床筛查的潜能。

鼻咽癌是东南亚地区及中国南方地区高发的恶性肿瘤, 具有独特的地理、 种族分布特点, 并且存在恶性程度高、 预后差、 早期诊断困难等问题。 拉曼光谱技术是基于非弹性光散射基本原理的一种快速且无损的检测方法, 能够在分子振动水平上提供生化成分等信息。 综述基于拉曼光谱技术的鼻咽癌组织研究最新进展。 主要介绍了国内外小组采用拉曼光谱及表面增强拉曼光谱（SERS）进行鼻咽癌组织检测研究的概况, 其中重点介绍该研究小组近期在鼻咽癌组织的高波数拉曼光谱、 鼻咽癌组织涂片的拉曼光谱, 以及研发的人活体鼻咽癌组织内镜检测装置及其临床实验情况。 最后, 对鼻咽癌组织拉曼光谱研究的发展前景进行了展望。

本文以鼻咽癌细胞株CNE2为放射敏感性的研究对象,经不同剂量X射线照射及不同时间培养后分别提取总蛋白,用共聚焦显微拉曼光谱仪检测其拉曼光谱.统计分析表明:被测样品的拉曼光谱中观察到一些可以归属于蛋白质物质的较为明显的基团频率振动峰;不同剂量的X射线照射后,总蛋白质的平均拉曼光谱与对照组谱形基本一致,但与对照组间的光谱存在着对应峰信号强度的不同.实验提示:照射后谱峰强度的增大或减小,提示着相关物质含量有所改变.分析照射后癌细胞总蛋白拉曼光谱的变化情况,结合数学统计方法,以探寻放射敏感性的特征拉曼标志,可以作为研究肿瘤放射敏感性的手段之一.

利用激光扫描共聚焦显微镜动态研究大黄素β环糊精包合物在鼻咽癌CNE-1细胞的跨膜转运及分布。结果显示，大黄素β环糊精包合物以颗粒的形式主要分布于胞浆中，在其浓度为5~40 mg·L-1时细胞对其摄取量随浓度的增加呈非线性增加；NaN3、甘露醇可抑制CNE-1细胞对大黄素β环糊精包合物的摄入量，环孢菌素A(CsA)在药物浓度低(小于5 mg·L-1)时可显著增加细胞对大黄素β环糊精包合物的摄入量，而在药物浓度大时，反而抑制细胞的摄取量。相对大黄素而言，大黄素β环糊精包合物在细胞内持续时间较长。大黄素β环糊精包合物在鼻咽癌CNE-1细胞的跨膜转运主要遵循能量依赖的内吞模式且受P-gp蛋白的调控，这一特性可为药物剂型研究提供参考。

拉曼光谱技术在肿瘤与正常细胞株的鉴别方面有着广泛的应用。 对一个已有的诊断模型进行可靠性验证是非常重要和有意义的工作。 采用两种不同的支持向量机分类算法对鼻咽癌和正常鼻咽细胞株的拉曼光谱进行分析和识别, 结果显示灵敏度和特异性均在90%以上, 并且与已知的相关线性判别分析结果一致。 结论表明, 两种支持向量机算法都能较好地对细胞株进行鉴别, 同时也表明拉曼光谱技术结合相关统计分类算法的方法可以实现对肿瘤细胞的准确鉴别, 这一结果将进一步证实拉曼光谱可以作为鼻咽癌诊断的一种方式。

Background and aims: The spectral properties of enhanced green fluorescent protein (EGFP) used in current visualizable animal models for nasopharyngeal carcinoma (NPC) result in a limited imaging depth. Far-red fluorescent proteins have optimal spectral wavelengths that allow deep tissue penetration, thus are well-suited for the imaging of tumor growth and metastases in live animals. This study aims to establish an imageable animal model of NPC using far-red fluorescent proteins. Methods: Eukaryotic expression vectors of far-red fluorescent proteins, mLumin and Katushka S158A, were separately transfected into 5-8F NPC cells, and cell lines stably expressing the far-red fluorescent proteins were obtained. These cells were intraperitoneally or intravenously injected into mice, and their tumorigenic and metastatic potential were examined through fluorescence imaging. Finally, factors affecting their tumorigenic ability were further assessed through testing side population (SP) cells proportion by flow cytometry. Results: NPC cell line with high tumorigenicity and metastasis (5-8F-mL2) was screened out, which stably expressed far-red fluorescent protein. Intraperitoneal and intravenous injection of 5- 8F-mL2 cells resulted in an abdomen metastasis model and a lung metastasis model. In addition, NPC cell line without tumorigenicity (5-8F-Katushka S158A) was screened out. The percentage of SP cells between 5-8F-mL2 and 5-8F-Katushka S158A was found different, suggesting that the SP cell proportion may play a key role in the determination of cell tumorigenic ability. Conclusion: We successfully established animal models for NPC with high tumorigenicity and metastasis using a super-bright far-red fluorescent protein. Owing to the super-brightness and excellent wavelength parameters, these models may be applied as useful tools for intuitive and efficient monitoring of tumor growth and metastasis, as well as assessing the efficacy of nasopharyngeal cancer drugs.

Nasopharyngeal carcinoma (NPC) is a prevalent cancer in some areas of southern Asia. To explore the potential of photodynamic therapy (PDT) for the treatment of NPC, a small molecule prodrug 5-aminolevulinic acid (ALA) and its methyl ester (MAL) mediated PDT was studied in vitro. The results showed that human NPC cells were sensitive to both ALA- and MAL-mediated PDT. However, ALA was more effective than MAL, possiblly due to a higher efficiency of ALA on producing endogenous protoporphyrin (PpIX) in NPC cells. Neither ALA nor MAL caused any significant genotoxicity. The ALA-based PDT might be a useful modality in the treatment ofNPC.

高空间分辨率的共聚焦显微拉曼光谱技术从分子水平上提供了生物分子成分和结构的指纹性信息。以鼻咽癌细胞株C666-1、CNE2和正常鼻咽细胞株NP69为对象，利用拉曼光谱探索和分析癌细胞株与正常细胞株之间的差别。结果表明：谱峰比I1449/I1657(1.10)可以准确地将鼻咽癌与正常鼻咽细胞株分成两组，这一结论与相关支气管癌组织的报道结果一致；此外，采用主成分分析方法说明3种细胞株的拉曼光谱之间存在统计学差异，并以线性判别分析方法对其进行统计划分，灵敏度和特异性分别达到90%和100%。这些结论可以为拉曼光谱技术用于NPC的临床诊断提供依据，并有可能为鼻咽癌的早期诊断提供有益参考。