量子点荧光探针检测人舌鳞状细胞癌荷瘤裸鼠模型早期下颌下淋巴结转移的研究

发布时间：2018-03-25 05:30

本文选题：舌鳞状细胞癌　切入点：量子点　出处：《华西口腔医学杂志》2014年03期

【摘要】：目的利用量子点标记的特异性细胞角蛋白抗体QDs605-CK(AE1/AE3)免疫荧光探针检测人舌鳞状细胞癌荷瘤裸鼠早期下颌下淋巴结转移率及微转移率,并与传统的免疫组织化学(IHC)染色及苏木精-伊红(HE)染色方法进行比较,为舌鳞状细胞癌的早期诊断与治疗提供一种新的检测方法。方法传代培养人舌鳞状细胞癌Tca8113细胞,接种于18只裸鼠舌体内(不过中线),建立人舌癌荷瘤裸鼠下颌下淋巴结转移模型。接种6周后,处死裸鼠,解剖下颌下淋巴结,将同一淋巴结分为两份。一份作石蜡包埋半连续切片,行HE染色和IHC检测;另一份即刻液氮冷冻,制作冰冻切片行QDs605-CK(AE1/AE3)荧光探针检测。分别计算3种方法检测出的淋巴结转移率和微转移率。结果量子点标记的免疫荧光染色检测出裸鼠下颌下淋巴结转移率为66.7%,其中微转移率为38.9%;IHC染色检测的淋巴结转移率为61.1%,其中微转移率为33.3%;HE染色检测的淋巴结转移率为27.8%。经统计学分析,3种方法的差异有统计学意义(χ2=6.379,P0.05),量子点标记的免疫荧光染色和IHC检测都优于HE染色,但是量子点标记的免疫荧光染色和IHC染色间的差异无统计学意义(χ2=0.120,P0.05)。结论量子点标记的QDs605-CK(AE1/AE3)免疫荧光探针能准确定位于下颌下淋巴结转移的肿瘤细胞的细胞质内,发出红色荧光,其特异性强,分辨率高,背景清晰,能够用于淋巴结转移灶及微转移灶的检测。
[Abstract]:Objective to detect the submandibular lymph node metastasis rate and micrometastasis rate in nude mice bearing human tongue squamous cell carcinoma by using QDs605-CKAE1 / AE3 immunofluorescence probe labeled with QDS-605-CKAE3 specific cytokeratin antibody (QDs605-CKAE1 / AE3). Compared with the traditional immunohistochemical staining and hematoxylin-eosin (HEH) staining, a new method for the early diagnosis and treatment of tongue squamous cell carcinoma (TSCC) was provided. Methods Human tongue squamous cell carcinoma (Tca8113) cells were subcultured. The submandibular lymph node metastasis model of human tongue cancer bearing nude mice was established by inoculating in 18 nude mice tongue (but midline). Six weeks after inoculation, nude mice were killed and submandibular lymph nodes were dissected. The same lymph node was divided into two parts. One was paraffin-embedded semi-continuous section, HE staining and IHC detection, and the other was immediately frozen liquid nitrogen. Frozen sections were made and detected with QDs605-CKAE1 / AE3 fluorescence probe. The lymph node metastasis rate and micrometastasis rate detected by three methods were calculated respectively. Results the submandibular lymph node metastasis rate of nude mice detected by quantum dot immunofluorescence staining was 66.7%, and that of QDs605-CKAE1 / AE3 fluorescence probe was determined by QDs605-CKAE1 / AE3 fluorescence probe. The median micrometastasis rate was 38.9% and the lymph node metastasis rate detected by IHC staining was 61.1%, of which the micrometastasis rate was 33.3% and the lymph node metastasis rate detected by HE staining was 27.80.The difference among the three methods was statistically significant (蠂 26.379U P0.05G, QDS-labeled immunofluorescence). Light staining and IHC detection were better than HE staining. But there was no significant difference between QDS-labeled immunofluorescence staining and IHC staining (蠂 ~ 2 = 0.120, P 0.05). Conclusion the QDs605-CKAE1 / AE3 probe can accurately locate in the cytoplasm of tumor cells with submandibular lymph node metastasis and emit red fluorescence. It has strong specificity, high resolution and clear background. It can be used to detect lymph node metastasis and micrometastasis.
【作者单位】：广东省深圳市宝安区妇幼保健院口腔科;广东医学院附属医院口腔科;
【基金】：湛江市科技专项分配基金资助项目(201020301)
【分类号】：R739.8

【参考文献】