鼻咽癌组织及细胞的拉曼光谱检测与分析
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摘要
鼻咽癌(Nasopharyngeal Carcinoma, NPC)是危害人类健康和生命的头颈部癌症之一,其发病率居耳鼻喉科恶性肿瘤之首。长期以来鼻咽癌诊断漏诊率较高,早期特别是Ⅰ期诊断率不高,而早期诊断后进行有效治疗是提高鼻咽癌患者生存率和降低死亡率的关键因素之一。本文利用拉曼光谱技术针对鼻咽癌组织、鼻咽正常组织及三种不同种类的鼻咽癌细胞开展一系列的拉曼光谱测量与分析,探讨鼻咽组织癌变过程中生化物质组成的变化,进而寻求实现鼻咽癌早期诊断的新方法,并在此基础上结合当今技术发展及临床需求,开展活体生物组织拉曼光谱测量装置的研究。
本文首先简要介绍了拉曼光谱的基本原理；综述了拉曼光谱技术在生物医学中的研究进展,并针对拉曼光谱数据处理方法进行了较为详细的分析与讨论。
其次,利用显微拉曼光谱仪及一套自行研制的快速拉曼光谱检测系统针对鼻咽癌组织、鼻咽正常组织及鼻咽癌细胞开展拉曼光谱测量与分析,分析鼻咽组织癌变过程中生化物质组成的变化,并主要通过主成分分析和判别分析构建判别算法,对鼻咽癌变组织与鼻咽正常组织以及三种不同种类的细胞进行判别,探讨正常组织与癌变组织以及不同细胞间的差别。
最后,利用一台半导体激光器、光栅光谱仪、CCD探测器及鼻咽内窥镜,设计、组装了一套可能应用于生物组织活体测量的拉曼光谱快速检测系统,并在小鼠模型上进行尝试。初步结果表明,该系统可在短时间内(3-5s)获得活体组织的高质量拉曼光谱信号,具有较高的应用潜力。
研究结果表明,拉曼光谱技术在区分鼻咽正常组织和鼻咽癌变组织以及不同种类的鼻咽细胞等方面具有巨大的潜力,所构建的快速拉曼光谱检测系统可用于活体组织测量。本文研究结果将为拉曼光谱在鼻咽癌早期检测进而拓展拉曼光谱在生物医学中的应用提供重要的实验和理论依据。
Nasopharyngeal malignancies remain one of the major causes of cancer associated death and have high incidence rates in East Asia, particularly in South China. Presently, routine white-light endoscopy and histological examination are the primary methods for clinical identification of nasopharyngeal carcinoma (NPC). Due to the anatomical position of NPC, it is not easy to diagnosis this lesion early, only about10%and20%of the diagnosed lesions are stage Ⅰ and Ⅱ diseases. Almost two thirds of the diagnosed nasopharyngeal carcinoma are at an advanced stage (stage Ⅲ). Due to the fact that the therapeutic outcome is highly related to the stage of the disease, early identification of malignant nasopharyngeal carcinoma is crucial to improving the survival rate of patients.
Recent development of spectral techniques may significantly expand our ability to diagnose this tumor rapidly and accurately. Among the optical approaches currently under investigation for in vivo endoscopic applications, Raman spectroscopy is a very promising technique. Raman spectroscopy is a non-destructive, inelastic light scattering technique in which the scattered photon is shifted to another wavelength with respect to the incident excitation light, depending on the specific vibrational modes of molecules in tissue and cells. Thus Raman spectroscopy reveals specific biochemical information and biomolecular structures of tissue, providing the unique opportunity to distinguish between different pathological tissue types at the molecular level.
In this paper Raman spectroscopy was used to detect the signals of normal nasopharyngeal tissue and nasopharyngeal carcinoma tissue as well as three different nasopharyngeal cell lines such as C666, CNE2and NP69. According to the differences of Raman spectroscopy characterization, we are intent to search for a new method which can diagnose nasopharyngeal carcinoma at its early stage. Combined with the new fiber and LD technologies, new instrument of Raman system for in vivo tissue detection was also studied.
In the first part of this paper, fundamental theory of Raman spectroscopy was introduced and a brief review of Raman applications in biomedical field was summarized. Then, Raman spectra of nasopharyngeal carcinoma tissue and normal tissue as well as three different cell lines were acquired using a micro-Raman system and a home-made rapid Raman system, respectively. The differences of Raman spectroscopy characterization were analyzed by comparing their spectroscopic intensity and tentative assigned to biochemical components. For further understanding the differences between two different tissues, PCA and LDA was adopted which yielded a diagnostic sensitivity and specificity over than80%.
In the end, an endoscopy based rapid Raman detection system was designed and built which is suitable for in vivo measurement. Raman spectra of SCC rat skin tissue were measured and the results have shown the system is reliable and can acquire high quality Raman signals which has a great potential application for in vivo human tissue measurement. All the preliminary results have suggest great potential for using Raman spectroscopy to improve the diagnosis of nasopharyngeal cancer. It provides solid support for us to develop endoscopic Raman instrument for in vivo clinical applications.
本文首先简要介绍了拉曼光谱的基本原理；综述了拉曼光谱技术在生物医学中的研究进展,并针对拉曼光谱数据处理方法进行了较为详细的分析与讨论。
其次,利用显微拉曼光谱仪及一套自行研制的快速拉曼光谱检测系统针对鼻咽癌组织、鼻咽正常组织及鼻咽癌细胞开展拉曼光谱测量与分析,分析鼻咽组织癌变过程中生化物质组成的变化,并主要通过主成分分析和判别分析构建判别算法,对鼻咽癌变组织与鼻咽正常组织以及三种不同种类的细胞进行判别,探讨正常组织与癌变组织以及不同细胞间的差别。
最后,利用一台半导体激光器、光栅光谱仪、CCD探测器及鼻咽内窥镜,设计、组装了一套可能应用于生物组织活体测量的拉曼光谱快速检测系统,并在小鼠模型上进行尝试。初步结果表明,该系统可在短时间内(3-5s)获得活体组织的高质量拉曼光谱信号,具有较高的应用潜力。
研究结果表明,拉曼光谱技术在区分鼻咽正常组织和鼻咽癌变组织以及不同种类的鼻咽细胞等方面具有巨大的潜力,所构建的快速拉曼光谱检测系统可用于活体组织测量。本文研究结果将为拉曼光谱在鼻咽癌早期检测进而拓展拉曼光谱在生物医学中的应用提供重要的实验和理论依据。
Nasopharyngeal malignancies remain one of the major causes of cancer associated death and have high incidence rates in East Asia, particularly in South China. Presently, routine white-light endoscopy and histological examination are the primary methods for clinical identification of nasopharyngeal carcinoma (NPC). Due to the anatomical position of NPC, it is not easy to diagnosis this lesion early, only about10%and20%of the diagnosed lesions are stage Ⅰ and Ⅱ diseases. Almost two thirds of the diagnosed nasopharyngeal carcinoma are at an advanced stage (stage Ⅲ). Due to the fact that the therapeutic outcome is highly related to the stage of the disease, early identification of malignant nasopharyngeal carcinoma is crucial to improving the survival rate of patients.
Recent development of spectral techniques may significantly expand our ability to diagnose this tumor rapidly and accurately. Among the optical approaches currently under investigation for in vivo endoscopic applications, Raman spectroscopy is a very promising technique. Raman spectroscopy is a non-destructive, inelastic light scattering technique in which the scattered photon is shifted to another wavelength with respect to the incident excitation light, depending on the specific vibrational modes of molecules in tissue and cells. Thus Raman spectroscopy reveals specific biochemical information and biomolecular structures of tissue, providing the unique opportunity to distinguish between different pathological tissue types at the molecular level.
In this paper Raman spectroscopy was used to detect the signals of normal nasopharyngeal tissue and nasopharyngeal carcinoma tissue as well as three different nasopharyngeal cell lines such as C666, CNE2and NP69. According to the differences of Raman spectroscopy characterization, we are intent to search for a new method which can diagnose nasopharyngeal carcinoma at its early stage. Combined with the new fiber and LD technologies, new instrument of Raman system for in vivo tissue detection was also studied.
In the first part of this paper, fundamental theory of Raman spectroscopy was introduced and a brief review of Raman applications in biomedical field was summarized. Then, Raman spectra of nasopharyngeal carcinoma tissue and normal tissue as well as three different cell lines were acquired using a micro-Raman system and a home-made rapid Raman system, respectively. The differences of Raman spectroscopy characterization were analyzed by comparing their spectroscopic intensity and tentative assigned to biochemical components. For further understanding the differences between two different tissues, PCA and LDA was adopted which yielded a diagnostic sensitivity and specificity over than80%.
In the end, an endoscopy based rapid Raman detection system was designed and built which is suitable for in vivo measurement. Raman spectra of SCC rat skin tissue were measured and the results have shown the system is reliable and can acquire high quality Raman signals which has a great potential application for in vivo human tissue measurement. All the preliminary results have suggest great potential for using Raman spectroscopy to improve the diagnosis of nasopharyngeal cancer. It provides solid support for us to develop endoscopic Raman instrument for in vivo clinical applications.
引文
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