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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

Application of fluorescence probe marked by quantum dots to detect early submandibular lymph node metastasis in a nude mouse model
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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,P<0.05),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,P>0.05)。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。 Objective A fluorescence probe marked with a specific angle protein antibody by using quantum dots QDs605-CK (AE1/AE3) was utilized to detect metastasis and micrometastasis rates of early submandibular lymph node metastatic carcinoma in a nude mouse model. The method was compared with traditional immunohistochemistry (IHC) staining and hematine-eosin (HE) staining to explore a new method for early diagnosis and treatment of tongue squamous cell carcinoma. Methods Human tongue Tca8113 cells were cultured and inoculated in 18 nude mice tongue body tissues (except the midline) to establish submandibular lymph node metastasis in tumor-bearing nude mice models. The nude mice were sacrificed, and the mandibular lymph node was dissected after injecting tumor cells for six weeks. One lymph node was divided into two parts: one was used to create a serial section embedded by paraffin then detected in HE staining and IHC staining; the other was immediately frozen in liquid nitrogen and detected using the fluorescence probe marked with a specific angle protein antibody by using quantum dots. Lymph node metastasis and micrometastases rates were calculated. Results The rates of metastasis in the submandibular lymph node via quantum dot immunofluorescence staining, IHC staining, and HE staining showed positive values of 66.7%, 61.1%, and 27.8%, res-pectively. The micrometastases rates were 38.9% and 33.3% via quantum dot immunofluorescence staining and IHC staining, respectively. The positive rates of detecting lymph nodes metastasis with quantum dot labeled keratin CK (AE1/AE3) immunofluorescence and keratin CK (AE1/AE3) IHC staining were higher than that of conventional HE staining (χ2=6.379, P〈0.05). The two methods showed no statistical diffe-rence (χ2=0.120, P〉0.05). Conclusion Quantum dots fluorescent probe marked with the angle protein antibody CK (AE1/AE3) can precisely locate the submandibular lymph node metastasis tumor cell of the nude mice tongue squamous cell car-cinoma, and the emitted red fluorescence showed strong specificity, high resolution, and a clear background. The method could be utilized to detect submandibular lymph node metastasis and micrometastasis of tongue squamous cell carcinoma in nude mice.
作者 李艳芬 陈坤
出处 《华西口腔医学杂志》 CAS CSCD 北大核心 2014年第3期273-277,共5页 West China Journal of Stomatology
基金 湛江市科技专项分配基金资助项目(201020301)
关键词 舌鳞状细胞癌 量子点 下颌下淋巴结 微转移 细胞角蛋白 tongue squamous carcinoma quantum dots submandibular lymph nodes micrometastasis cytokeratin
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