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下咽癌颏下区成瘤与腋下区成瘤裸鼠模型的比较
闵密克,周鹏,朱敏辉,宋先敏,郑宏良*
0
(海军军医大学(第二军医大学)长海医院耳鼻咽喉头颈外科, 上海 200433
*通信作者)
摘要:
目的 建立一种下咽癌颏下区成瘤裸鼠模型,并与传统腋下区成瘤裸鼠模型进行比较。方法 选择5周龄BALB/c雄性裸鼠,分为颏下区成瘤裸鼠模型组(颏下组)和腋下区成瘤裸鼠模型组(腋下组)。每组又分为两个亚组,分别注射人下咽癌细胞株FADU及HN31,各亚组12只。颏下组于左颏下区注射肿瘤细胞,腋下组于右腋下注射肿瘤细胞,记录4个亚组裸鼠成瘤时间、成瘤体积、体质量。记录裸鼠死亡情况。对8周后仍生存的裸鼠行超声检查,观察局部肿瘤生长、浸润情况及肝、脾、肾等器官转移情况,并行常规H-E染色和免疫组织化学染色观察肿瘤的转移情况。结果 接种后第7天时颏下组裸鼠即有瘤体出现,第9天时腋下组出现瘤体;第33天时FADU腋下组、HN31腋下组裸鼠瘤体体积分别大于FADU颏下组、HN31颏下组(P<0.05,P<0.01)。各组裸鼠体质量至接种后第17~19天基本达到峰值,此后颏下组裸鼠体质量逐步减轻,腋下组仍有所增加;第33天时FADU腋下组、HN31腋下组裸鼠体质量分别大于FADU颏下组、HN31颏下组(P均<0.01)。第8周时FADU颏下组裸鼠死亡8只,HN31颏下组死亡10只,颏下组共存活6只;FADU腋下组死亡6只,HN31腋下组死亡6只,腋下组共存活12只。不同接种部位、接种不同细胞组间裸鼠死亡率差异均无统计学意义。超声和病理检查结果显示,颏下组6只存活裸鼠中,4只发现颈部淋巴结转移及肝脏转移灶,腋下组8只存活裸鼠中仅1只发现肝脏转移灶,两组间差异有统计学意义(P<0.05)。颏下组有2只发现肺部转移灶,腋下组有1只发现肺部转移灶;颏下组有1只发现脾脏转移灶,腋下组无裸鼠存在脾脏转移灶,两组间差异均无统计学意义。结论 颏下区成瘤与腋下区成瘤裸鼠模型各有特点。颏下区成瘤时间短,局部侵害重、远处转移比例高,适用于研究肿瘤在体侵袭性和转移性。腋下区成瘤可形成较大体积瘤体,但邻近器官、组织受累程度轻,裸鼠存活时间长,但远处转移率相对较低,适用于肿瘤细胞自身特性的研究。
关键词:  下咽肿瘤  颏下区  腋下区  实验性肿瘤  裸小鼠
DOI:10.16781/j.0258-879x.2018.06.0597
投稿时间:2018-04-02修订日期:2018-06-12
基金项目:国家自然科学基金(81572668,81402253).
Comparison of submental tumorigenesis model and axillary tumorigenesis model in nude mice xenotransplanted with human hypopharyngeal carcinoma cells
MIN Mi-ke,ZHOU Peng,ZHU Min-hui,SONG Xian-min,ZHENG Hong-liang*
(Department of Otolaryngology/Head and Neck Surgery, Changhai Hospital, Navy Medical University(Second Military Medical University), Shanghai 200433, China
*Corresponding author)
Abstract:
Objective To establish a submental tumorigenesis model by injecting human hypopharyngeal carcinoma cells to nude mice, and to compare the model with traditional axillary tumorigenesis model. Methods Five-week-old male nude mice were selected and divided into submental tumorigenesis model group (submental group) and axillary tumorigenesis model group (axillary group). Each group was divided into two subgroups by injecting human hypopharyngeal carcinoma cells FADU or HN31, namely sub-FADU, sub-HN31, ax-FADU, and ax-HN31 groups, with 12 nude mice in each group. The nude mice in the submental group were injected with tumor cells in the left side of submental area, and those in the axillary group were injected with tumor cells in the right side of axillary area. The time of tumor formation, tumor volume and body mass of nude mice were measured. The death of nude mice was recorded. After eight weeks, the local tumor growth, infiltration, and organ metastasis such as liver, spleen and kidney of the survival nude mice were examined by ultrasound, and the tumor metastasis was observed by routine H-E staining and immunohistochemical staining. Results On the 7th and 9th days after injection, the tumors were observed in the submental group and the axillary group. On the 33rd day after injection, the tumor volume in the ax-FADU and ax-HN31 groups was significantly bigger than those in the sub-FADU and sub-HN31 groups (P<0.05, P<0.01). The body mass of nude mice in each group reached peak on the 17th to 19th days after injection, and thereafter the body mass in the submental group was gradually decreased and that in the axillary group was still increased. On the 33rd day after injection, the body mass in the ax-FADU and ax-HN31 groups were significantly higher than those in the sub-FADU and sub-HN31 groups, respectively (P<0.01). After eight weeks, there were eight deaths in the sub-FADU group and 10 in the sub-HN31 group, with six mice alive in the submental group; there were six deaths in the ax-FADU group and six in the ax-HN31 group, with 12 alive in the axillary group. There was no significant difference in mortality of nude mice between the four subgroups. Ultrasound and pathological examination showed that four survived nude mice were found with cervical lymph node and liver metastases in the submental group, and only one was found with liver metastases in the axillary group; and the difference between the two groups was statistically significant (P<0.05). There were two pulmonary metastases in the submental group, and one in the axillary group; there was one spleen metastases in the submental group and no spleen metastases in the submental group; and there were no significant differences between the two groups. Conclusion Submental tumorigenesis model and axillary tumorigenesis model have their own characteristics. The submental tumorigenesis model has shorter tumorigenesis time, higher local invasion and higher distant metastasis rate, and is suitable for studying the invasiveness and metastasis of tumor in vivo. The axillary tumorigenesis model has larger volume of tumor, less injury to the adjacent organs and tissues, longer survival time and lower distant metastasis rate, and is suitable for the study of the characteristics of tumor cells.
Key words:  hypopharyngeal neoplasms  submental region  axillary region  experimental neoplasms  nude mice