EGFRmAb功能化修饰金纳米棒光热诱导喉鳞癌细胞凋亡分子机制研究

发布时间：2018-07-09 19:22

  本文选题：喉癌 + 表皮生长因子受体　； 参考：《昆明医科大学》2012年博士论文

【摘要】：第一部分表皮生长因子受体在喉鳞癌组织中的表达及siRNA沉默EGFR基因诱导Hep-2细胞凋亡研究 目的：检测喉鳞癌组织中表皮生长因子受体(EGFR)的表达,探讨其在喉癌发生中的作用。运用RNA干扰技术研究EGFR表达下调诱导Hep-2细胞凋亡情况。 方法：收集2008年1月～2011年3月昆明医学院第一附属医院、第三附属医院头颈外科36例喉鳞癌组织标本,正常对照来自其中的13例喉全切除术标本,阳性对照选取Hep-2细胞株。运用Western Blot技术检测喉癌组织、癌旁组织及正常粘膜组织EGFR的表达,图像结果进行Quantity One软件定量分析,得出EGFR蛋白相对表达量,分析其与癌组织病理分级的关系。进而运用siRNA技术沉默Hep-2细胞EGFR基因,RT-PCR及Western Blot检测EGFR基因表达抑制效果,流式细胞术进一步检测EGFRsiRNA质粒转染48h时Hep-2细胞凋亡情况。 结果：(1)喉癌标本中EGFR蛋白的表达从高到低依次为喉鳞癌组织、癌旁组织,正常粘膜组织几乎无条带显示。喉癌组织、癌旁组织不同病理分级之间EGFR相对表达量差异有统计学意义,P0.05。相同病理分级喉癌组织、癌旁组织及正常组织之间EGFR相对表达量差异有高度统计学意义,P0.01。(2)成功获得EGFRsiRNA表达质粒pSi-hEGFR,转染Hep-2细胞48h后RT-PCR检测结果显示,EGFRmRNA表达水平下调46%,Western Blot检测获得一致的结果。pSi-hEGFR质粒转染Hep-2细胞48h后流式细胞术检测发现：EGFR阳性表达抑制率为83.8%；细胞凋亡率达(15.13±2.15)%,而pSi-Negative质粒转染组凋亡率为(7.03±1.08)%,差异有高度统计学意义,P0.01。 结论：EGFR在喉癌组织中存在过表达现象,与喉癌组织不同部位及病理分级有相关性。siRNA沉默EGFR基因可以下调Hep-2细胞EGFR mRNA和蛋白水平表达,同时发现可以诱导Hep-2细胞出现凋亡。本研究提示EGFR可能参与喉癌的发生,其表达下调可以起到部分抑瘤作用,为以EGFR为靶点的抗肿瘤治疗方法的探索提供基础理论依据。 第二部分EGFRmAb功能化修饰金纳米棒光热作用抑制Hep-2细胞增殖研究 目的：用EGFRmAb功能化修饰金纳米棒(AuNRs)获得稳定的EGFRmAb/AuNRs共轭物,探索AuNRs进入Hep-2细胞的机制,了解AuNRs光热转换的升温规律,评价AuNRs的生物安全性,观察EGFRmAb功能化修饰AuNRs靶向光热效应抑制Hep-2细胞增殖作用。 方法：合成EGFRmAb/AuNRs后用透射电镜进行表征,用紫外-可见-近红外光谱仪对其进行光谱分析。将AuNRs、EGFRmAb/AuNRs分别与Hep-2细胞共孵育,透射电镜观察AuNRs进入细胞及细胞内分布。采用ICP-AES元素分析法检测EGFRmAb介导下AuNRs进入Hep-2细胞的量。NIR激光照射后用温度计测量不同状况下AuNRs溶液温度上升规律。用MTT法检测不同浓度AuNRs细胞毒性。用台盼蓝染色法大体观察AuNRs光热作用杀伤肿瘤细胞现象,继而用MTT法检测不同功率NIR激光照射AuNRs后不同时间点Hep-2细胞的增殖抑制率,比较AuNRs和EGFRmAb/AuNRs光热作用的抑瘤效果。 结果：(1)电镜表征显示本实验用AuNRs具有良好的稳定性和分散性,微粒大小均匀,长径均值为49.81nm,横径均值为12.70nm,长横比均值为3.92。紫外-可见-近红外光谱仪检测显示,AuNRs溶液表现出双共振吸收峰,横向为510nm,纵向为800nm。AuNRs经EGFRmAb修饰后纵向共振吸收峰位红移7-9nm,并不明显改变原有光学特性。(2)电镜观察显示：AuNRs和Hep-2细胞共同孵育30min时,可见AuNRs于细胞膜表面,出现胞吞现象；3h时可见AuNRs成簇状位于溶酶体内；6h时可见AuNRs同时存在于细胞质、溶酶体和线粒体内,少量AuNRs贴近核膜；未发现细胞形态和细胞器(线粒体)结构明显改变。EGFRmAb功能化修饰的AuNRs进入细胞的数量比未修饰的要多,进一步ICP-AES元素分析显示共孵育6h时EGFRmAb功能化修饰明显提高AuNRs进入细胞的数量,增加48.14%。(3)NIR激光照射AuNRs溶液可以引起温度迅速升高,温度升高的幅度和AuNRs浓度、照射功率及容器容积有一定关系。研究发现,在室温为23℃情况下,用3.0W/cm2功率NIR激光照射0.1nmol/L AuNRs溶液(1ml/每孔,24孔板)4min时温度达到46℃,随后稳定,满足本实验光热诱导凋亡的温度要求。(4)MTT法检测显示AuNRs在0.1nmol/L～2.0nmol/L浓度范围内未出现细胞活力明显下降,AuNRs/EGFRmAb共轭物对Hep-2细胞活力没有明显影响。(5)5.2W/cm2功率NIR激光照射AuNRs和Hep-2细胞共孵育培养液8min后台盼蓝染色显示细胞全部死亡。进一步MTT法检测显示：AuNRs和AuNRs/EGFRmAb光热作用可以明显引起Hep-2细胞增殖抑制,呈激光功率剂量依赖性；而单纯NIR激光照射及EGFR单抗对Hep-2细胞没有明显的增殖抑制作用；控制NIR激光功率为3.0W/cm2,则Hep-2细胞的增殖抑制呈明显的时间效应关系,72h抑制率最高,达69.24%；同时发现,AuNRs/EGFRmAb光热效应对Hep-2细胞的增殖抑制作用明显高于未修饰AuNRs, P0.01. 结论：EGFRmAb修饰AuNRs后未明显改变原有的形状和光学性质,但可以明显增加AuNRs进入Hep-2细胞的能力,推测可能和EGFR介导的胞吞有关。AuNRs进入细胞后主要存在于细胞质、溶酶体和线粒体中,其本身并没用明显的细胞毒性,但在NIR激光的照射下,能迅速引起环境温度的升高,导致细胞出现增殖抑制或死亡。控制NIR激光的照射功率,细胞的增殖抑制呈时效性。EGFRmAb功能化修饰明显提高AuNRs光热抑瘤效果,而单纯的NIR激光照射和EGFRmAb并没有明显的细胞增殖抑制现象。由于AuNRs位于细胞内线粒体中,我们推测上述细胞增殖抑制可能是AuNRs光热诱导的内源性凋亡导致的。 第三部分EGFRmAb/AuNRs光热诱导Hep-2细胞凋亡机制研究 目的：控制NIR激光照射参数,观察EGFRmAb/AuNRs光热诱导Hep-2细胞凋亡现象,通过检测线粒体途径凋亡相关蛋白分子表达来阐述凋亡分子机制,同时证明EGFRmAb功能化修饰能够有效提高AuNRs光热诱导Hep-2细胞凋亡效果。 方法：参照第二部分实验,控制NIR激光照射参数为：功率3.0W/cm2,时间8min。电镜观察照射后Hep-2细胞超微结构变化及凋亡。TUNEL/PI双染法流式细胞术检测照射后不同时间点细胞凋亡和周期,进而流式细胞术检测照射后不同时间点Hep-2细胞活化Caspase-3、Bcl-2/Bax和Cyt-c阳性表达率、细胞内ROS水平和Ca2+浓度及细胞线粒体膜电位(△甲m)下降率。 结果：(1)照射后48h电镜观察NIR+AuNRs组和NIR+EGFRmAb/AuNRs组可见细胞凋亡。(2)NIR+AuNRs组和NIR+EGFRmAb/AuNRs组细胞在照射后24h、48h时表现出明显的G0/G1期阻滞,同时S期比率下降,并呈时间效应关系。在照射后48h时,EGFRmAb/AuNRs组和AuNRs组相比,S期比率下降相对明显,P0.01。(3)NIR+AuNRs组与NIR+EGFRmAb/AuNRs组细胞在照射后出现明显凋亡,呈时间效应关系。NIR+EGFRmAb/AuNRs组细胞在照射后三个时间点(24h、48h、72h)的凋亡：率均比NIR+AuNRs组要高,P0.05。照射后72h时NIR+EGFRmAb/AuNRs组细胞凋亡率最高,达73.63%。(4)照射后NIR+AuNRs组和NIR+EGFRmAb/AuNRs组细胞活化Caspase-3、Bax阳性表达率、△Ψm下降率及细胞内ROS水平、Ca2+浓度升高,细胞Bcl-2阳性表达率降低。随时间推移上述指标各自变化趋势明显,但NIR+EGFRmAb/AuNRs组细胞上述指标改变比NIR+AuNRs组更为明显,P0.05。照射后NIR+AuNRs组与NIR+EGFRmAb/AuNRs组细胞Cyt-c阳性表达率明显升高。照射后1h时检出Cyt-c阳性表达,24h时最高,随后呈逐渐下降趋势。在照射后24h、48h时,NIR+EGFRmAb/AuNRs组细胞Cyt-c阳性表达率和NIR+AuNRs组比较升高更为明显,P0.01。 结论：控制合适的NIR激光照射参数可以通过AuNRs光热作用诱导Hep-2细胞出现凋亡,这种凋亡和细胞周期阻滞密切相关。ROS和Ca2+含量升高、△ψm下降、Cyt-c释放、活化Caspase-3上调及Bcl-2/Bax比例下降在上述凋亡过程中起重要作用,可以看出,线粒体介导的内源性途径是AuNRs光热诱导Hep-2细胞凋亡的重要方式。EGFRmAb功能化修饰可以通过AuNRs的靶向聚集而提高诱导凋亡的效果,但EGFR单抗本身并没有明显参与凋亡的发生。 第四部分EGFRmAb/AuNRs光热治疗裸鼠喉鳞癌移植瘤研究 目的：观察局部注射及全身静脉给药模式下AuNRs光热治疗裸鼠皮下喉癌移植瘤效果并检测治疗后肿瘤细胞凋亡情况。 方法：建立裸鼠皮下喉癌移植瘤模型。瘤体局部注射AuNRs,经尾静脉全身给EGFRmAb/AuNRs,NIR激光照射参数同第三部分体外实验,照射过程中测量肿瘤局部皮温变化,游标卡尺测量照射后肿瘤大小变化。照射后2周结束实验,绘制移植瘤生长曲线,裸鼠处死后切取肿瘤标本,TUNEL/PI双染法流式细胞术检测肿瘤细胞凋亡和坏死。 结果：NIR激光照射3min后肿瘤处皮肤温度趋于稳定,AuNRs局部注射组照射后温度上升15℃,EGFRmAb/AuNRs静脉给药组照射后温度上升11℃。两观察组照射后肿瘤的生长均有明显的抑制现象,各观察时间点生长速度明显变缓,但AuNRs局部注射组照射后肿瘤抑制效果更明显一些,P0.05。单独EGFRmAb(E3138,Sigma公司)静脉给药、小剂量NIR激光照射及不经EGFRmAb修饰的AuNRs静脉给药后照射均没有产生明显的肿瘤抑制效果。流式细胞术检测发现,AuNRs局部注射组及EGFRmAb/AuNRs静脉给药组照射后2周肿瘤组织细胞出现明显的凋亡,凋亡率分别为58.35%和30.46%。AuNRs局部注射组照射后凋亡率和坏死率均明显高于EGFRmAb/AuNRs静脉给药组,P0.01。 结论：AuNRs直接瘤体注射光热作用可以明显抑制裸鼠喉癌移植瘤的生长,EGFRmAb/AuNRs静脉给药也可以起到一定的治疗效果,流式细胞术检测印证了光热诱导凋亡现象的存在。动物实验的效果给EGFRmAb/AuNRs体内靶向光热治疗肿瘤带来希望,由于体内环境的复杂性,要真正实现体内深层肿瘤安全有效的AuNRs光热治疗还需进一步深入研究。
[Abstract]:Part one expression of epidermal growth factor receptor in laryngeal squamous cell carcinoma and apoptosis of Hep-2 cells induced by siRNA silencing EGFR gene
Objective: to detect the expression of epidermal growth factor receptor (EGFR) in the tissues of laryngeal squamous cell carcinoma (LSCC) and explore its role in the development of larynx cancer. RNA interference technique was used to study the down regulation of EGFR expression to induce apoptosis in Hep-2 cells.
Methods: from January 2008 to March 2011, 36 cases of laryngeal squamous cell carcinoma were collected from the First Affiliated Hospital of Kunming Medical University, third affiliated hospitals, and 13 cases of laryngeal squamous cell carcinoma were collected from normal controls. The positive controls were selected from 13 cases of total laryngectomy. Western Blot technique was used to detect the tissues of the larynx, the para cancerous tissue and the normal mucous tissue EGFR By quantitative analysis of Quantity One software, the relative expression of EGFR protein was obtained, and the relationship between EGFR protein and histopathological classification was analyzed. Then siRNA technique was used to silence Hep-2 cell EGFR gene, RT-PCR and Western Blot were used to detect the inhibitory effect of EGFR gene expression. Flow cytometry further detected EGFRsiRNA plasmid transfection 48h. Apoptosis of EP-2 cells.
Results: (1) the expression of EGFR protein in the laryngeal carcinoma specimens from high to low was in the order of laryngeal squamous cell carcinoma, the para cancerous tissue, and the normal mucosal tissue almost no strip display. The relative expression of EGFR in the larynx tissues and the para cancerous tissues was statistically significant. P0.05. was the same pathological classification of the larynx, para cancer tissue and normal tissue. The relative expression of EGFR was statistically significant. P0.01. (2) successfully obtained the EGFRsiRNA expression plasmid pSi-hEGFR. The RT-PCR detection results of 48h transfected Hep-2 cells showed that the expression level of EGFRmRNA was down down by 46%, and the Western Blot detection obtained the same result of the.PSi-hEGFR plasmid transfected to the Hep-2 cells after the flow cytometry. The inhibition rate of expression was 83.8%, the apoptosis rate was (15.13 + 2.15)%, while the apoptosis rate of pSi-Negative plasmid transfected group was (7.03 + 1.08)%, and the difference was statistically significant, P0.01.
Conclusion: there is a phenomenon of overexpression of EGFR in the carcinoma of the larynx..siRNA silencing EGFR gene can down regulate the expression of EGFR mRNA and protein in Hep-2 cells and induce apoptosis of Hep-2 cells. This study suggests that EGFR may be involved in the occurrence of laryngeal carcinoma and its expression can be down regulated. It plays a part of tumor suppressive effect, and provides a basic theoretical basis for the exploration of anti-tumor therapy targeting EGFR.
The second part of EGFRmAb functionalized gold nanorods inhibit the proliferation of Hep-2 cells by photothermal action.
Objective: to obtain stable EGFRmAb/AuNRs conjugates with EGFRmAb functionalized gold nanorods (AuNRs), explore the mechanism of AuNRs into Hep-2 cells, understand the heating law of AuNRs photothermal transformation, evaluate the biosafety of AuNRs, and observe the function of EGFRmAb functionalized modified AuNRs target to inhibit the proliferation of Hep-2 cells.
Methods: the EGFRmAb/AuNRs was characterized by transmission electron microscopy, and the spectral analysis was carried out by ultraviolet visible near infrared spectrometer. AuNRs, EGFRmAb/AuNRs were incubated with Hep-2 cells respectively. The distribution of AuNRs into cells and cells was observed by transmission electron microscopy. ICP-AES element analysis was used to detect AuNRs into Hep-2 cells under EGFRmAb. The temperature rise of AuNRs solution under different conditions was measured by a thermometer after.NIR laser irradiation. The cytotoxicity of AuNRs cells at different concentrations was detected by MTT method. The phenomenon of AuNRs photothermal effect on tumor cells was observed by trypan blue staining, and then MTT method was used to detect the proliferation of Hep-2 cells at different time points of different power NIR lasers at different time points. The inhibition rate was compared with the antitumor effect of AuNRs and EGFRmAb/AuNRs photothermal therapy.
Results: (1) the electron microscopic characterization showed that the experiment with AuNRs had good stability and dispersion, the particle size was uniform, the mean length of diameter was 49.81nm, the mean diameter of the transverse diameter was 12.70nm, the mean value of the long transverse ratio was 3.92. ultraviolet visible near infrared spectrometer, and the AuNRs solution showed the double resonance absorption peak, the transverse 510nm, and the longitudinal 800nm.AuNRs via EGFRm. The longitudinal resonance absorption peak was red shift 7-9nm after Ab modification. (2) electron microscope observation showed that when AuNRs and Hep-2 cells incubated 30min together, AuNRs was found on the surface of the cell membrane, and there was a phenomenon of endocytosis; AuNRs was clustered in the soluble enzyme in 3h, and AuNRs was found at the same time at the cytoplasm, lysosome and. In mitochondria, a small amount of AuNRs was close to the nuclear membrane; no cell morphology and organelles (mitochondria) structure obviously changed the number of.EGFRmAb functionalized AuNRs to enter cells more than that of unmodified ones. Further ICP-AES analysis showed that EGFRmAb functional modification significantly increased the number of AuNRs into cells and increased 48.14%. (3). NIR laser irradiation of AuNRs solution can cause a rapid increase in temperature. The amplitude of the temperature rise and the concentration of AuNRs, the power of the irradiation and the volume of the container have a certain relation. The study found that the temperature of 0.1nmol/L AuNRs solution (1ml/ per pore, 24 hole plate) at 4min at room temperature is 23 C at room temperature, and the temperature reaches 46 degrees C, then stable, and satisfies this. Temperature requirements for apoptosis induced by light and heat. (4) MTT assay showed that AuNRs did not markedly decrease cell viability in 0.1nmol/L ~ 2.0nmol/L concentration, and AuNRs/EGFRmAb conjugates had no significant influence on Hep-2 cell viability. (5) 5.2W/cm2 power NIR laser irradiation of AuNRs and Hep-2 cell co incubating medium was shown by 8min background blue staining All the cells died. Further MTT assay showed that the photothermal effect of AuNRs and AuNRs/EGFRmAb could obviously induce the proliferation inhibition of Hep-2 cells, and the laser power dose depended, while the pure NIR laser irradiation and the EGFR mAb did not inhibit the proliferation of Hep-2 cells, and the NIR laser power was 3.0W/cm2, then the Hep-2 cells increased. The inhibitory rate of 72h was the highest, up to 69.24%, and the inhibitory effect of AuNRs/EGFRmAb photothermal effect on the proliferation of Hep-2 cells was significantly higher than that of unmodified AuNRs, P0.01.
Conclusion: EGFRmAb modified AuNRs did not obviously change the original shape and optical properties, but it could significantly increase the ability of AuNRs to enter Hep-2 cells. It is presumed that EGFR mediated cytosol related.AuNRs enters the cell and is mainly present in the cytoplasm, lysosomes and mitochondria, and its body does not use obvious cytotoxicity, but in NIR laser Under the irradiation, it can cause the increase of the temperature of the environment, which leads to the proliferation inhibition or death of the cells. Control the power of NIR laser irradiation, the inhibitory effect of the cell proliferation on the aging.EGFRmAb functional modification obviously improves the effect of AuNRs photothermal inhibition, and the simple NIR laser irradiation and EGFRmAb have no obvious cell proliferation inhibition. Due to AuN Rs is located in the mitochondria of cells. We speculate that the above inhibition of cell proliferation may be caused by AuNRs photothermal induced endogenous apoptosis.
The third part is the mechanism of EGFRmAb/AuNRs photothermal induced apoptosis in Hep-2 cells.
Objective: to control the parameters of NIR laser irradiation, observe the apoptosis of Hep-2 cells induced by EGFRmAb/AuNRs photothermal, and to explain the molecular mechanism of apoptosis by detecting the expression of apoptosis related protein in mitochondrial pathway, and prove that the functional modification of EGFRmAb can effectively improve the effect of AuNRs photothermal induced apoptosis of Hep-2 cells.
Methods: with reference to the second parts of the experiment, the parameters of NIR laser irradiation were controlled: power 3.0W/cm2, time 8min. electron microscope observation of ultrastructural changes of Hep-2 cells and apoptosis and cycle of.TUNEL/PI double staining flow cytometry at different time points after irradiation, and then flow cytometry was used to detect Hep-2 cells at different time points after irradiation. Activation of Caspase-3, Bcl-2/Bax and Cyt-c positive expression rate, intracellular ROS level and Ca2+ concentration and cell mitochondrial membrane potential (Delta a m) decline rate.
Results: (1) the apoptosis of NIR+AuNRs and NIR+EGFRmAb/AuNRs groups was observed by 48h electron microscope after irradiation. (2) the cells in group NIR+AuNRs and NIR+EGFRmAb/AuNRs showed significant G0/G1 phase block at 24h and 48h after irradiation, and at the same time, the ratio of S phase decreased and showed a time effect relationship. When compared with the AuNRs group, the EGFRmAb/AuNRs group compared with the AuNRs group at 48h. The decrease of the rate was relatively obvious. The apoptosis of P0.01. (3) NIR+AuNRs group and NIR+EGFRmAb/AuNRs group was obvious after irradiation. The apoptosis rate of.NIR+EGFRmAb/AuNRs group cells in.NIR+EGFRmAb/AuNRs group after irradiation (24h, 48h, 72h) was higher than that of NIR+AuNRs group. The apoptotic rate of NIR+EGFRmAb/AuNRs group at 72h was the highest, up to 73 after P0.05. irradiation, reaching 73. After.63%. (4) irradiation, the cells in group NIR+AuNRs and group NIR+EGFRmAb/AuNRs activated Caspase-3, the positive expression rate of Bax, the decreasing rate of M and the ROS level in the cells, the increase of Ca2+ concentration and the decrease of the positive expression rate of the cells. The change trend of the above indexes was obvious with the time lapse, but the changes of the above indexes in the NIR+ EGFRmAb/AuNRs group were more than those of the NIR+AuNRs group. The positive expression rate of Cyt-c in the cells of group NIR+AuNRs and NIR+EGFRmAb/AuNRs was obviously increased after P0.05. irradiation. The positive expression of Cyt-c was detected at 1h after irradiation, and the highest in 24h and then gradually decreased. The Cyt-c positive expression rate of the NIR+EGFRmAb/AuNRs group was more obvious than that of the NIR+AuNRs group at 24h and 48h after irradiation.
Conclusion: the control of appropriate NIR laser irradiation parameters can induce apoptosis of Hep-2 cells by AuNRs photothermal effect. This apoptosis and cell cycle arrest are closely related to the increase of.ROS and Ca2+ content, delta m decline, Cyt-c release, activation of Caspase-3 up regulation and the decrease of Bcl-2/Bax ratio play an important role in the process of apoptosis, can be seen, line The endogenous pathway mediated by particles is an important way of AuNRs photothermal induced apoptosis of Hep-2 cells..EGFRmAb functional modification can improve the effect of apoptosis through the targeting of AuNRs, but the EGFR monoclonal antibody itself does not participate in the apoptosis.
The fourth part of EGFRmAb/AuNRs photothermal therapy for transplanted tumor of laryngeal squamous cell carcinoma in nude mice
Objective: To observe the effect of AuNRs photothermal therapy in the treatment of subcutaneous laryngeal cancer xenografts in nude mice under local injection and systemic intravenous administration, and to detect the apoptosis of tumor cells after treatment.
Methods: a tumor model of subcutaneous laryngocarcinoma in nude mice was established. The tumor body was partially injected with AuNRs, through the whole body of the tail vein to EGFRmAb/AuNRs, the parameters of the NIR laser irradiation and the third part of the experiment in vitro. The local skin temperature changes were measured during the irradiation. The vernier caliper was used to measure the size of the tumor after irradiation. The experiment was completed at the end of the irradiation for 2 weeks to draw the growth of the transplanted tumor. Tumor cells were harvested after nude mice were sacrificed. Apoptosis and necrosis of tumor cells were detected by TUNEL/PI double staining flow cytometry.
Results: the temperature of the skin tended to be stable after the NIR laser irradiation for 3min. The temperature of the AuNRs local injection group increased by 15 degrees C, the temperature of the EGFRmAb/AuNRs intravenous administration group increased by 11 degrees C. The growth of the tumor in the observation group was obviously inhibited, and the growth rate of the observation time points was obviously slowed, but the AuNRs local injection group The tumor suppressor effect was more obvious after irradiation. P0.05. alone EGFRmAb (E3138, Sigma) intravenous administration, low dose NIR laser irradiation and non EGFRmAb modified AuNRs intravenous administration did not produce significant tumor inhibition effect. Flow cytometry detection, AuNRs local injection group and EGFRmAb/AuNRs intravenous administration group irradiation After 2 weeks, the tumor tissue cells showed obvious apoptosis. The apoptosis rate and the necrosis rate of 58.35% and 30.46%.AuNRs local injection groups were significantly higher than that of the EGFRmAb/AuNRs intravenous group, P0.01.
Conclusion: the effect of AuNRs direct injection of light and heat can obviously inhibit the growth of xenograft in nude mice. EGFRmAb/AuNRs intravenous administration can also play a certain therapeutic effect. Flow cytometry shows the existence of photothermal induced apoptosis. The effect of animal experiments on EGFRmAb/ AuNRs target photothermal treatment of tumor brings hope to the tumor. Because of the complexity of the internal environment, it is necessary to further study the safe and effective AuNRs photothermal therapy for deep tumors in vivo.
【学位授予单位】：昆明医科大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R739.65

【参考文献】

相关期刊论文 前3条

1 刘爱红;孙康宁;李爱民;;肿瘤热疗机制与方法的研究进展[J];现代生物医学进展;2006年11期

2 彭远飞;郑民华;;肿瘤热疗的细胞分子作用机制及应用进展[J];世界华人消化杂志;2007年12期

3 廖遇平;靶向热消融治疗恶性肿瘤的现状[J];中国现代医学杂志;2004年03期

，

本文编号：2110405