喉癌动物模型中播散肿瘤细胞与突变型p53的研究

The study of disseminated tumor cell(DTC)and p53 gene in mouse model of laryngocarcinoma

目的:建立喉癌裸鼠肿瘤模型,进行外周血播散肿瘤细胞(Disseminated tumor cell,DTC)和肿瘤组织突变型p53基因的检测,判断两者之间的相关性及早期预示肿瘤远处转移和预后的价值。方法:从50只裸鼠中随机选择40只建立裸鼠喉癌肿瘤模型,其余10只为对照组,采用套式PCR法,以CK19 mRNA为标记物于肿瘤种植成功后第10周检测肿瘤组及对照组裸鼠外周血中的DTC;以RT-PCR和免疫组化法检测肿瘤组织中突变型p53蛋白表达情况。待裸鼠死亡后解剖肝、肺及淋巴结,采用连续病理切片HE染色证实是否发生转移。结果:肿瘤组DTC阳性率为50.0%(20/40),对照组为阴性,差异有统计学意义(P<0.05)。肿瘤组织中突变型p53阳性率为55.0%(22/40),p53的表达与DTC呈正相关。1年内DTC与突变型p53都为阳性组中位生存期为34周,阴性组为43周,生存差异有统计学意义(P<0.05);阳性组发生远处器官转移率为38.9%(7/18),阴性组转移率12.5%(2/16),差异有统计学意义(P<0.05)。结论:DTC联合突变型p53蛋白阳性表达可以作为早期判断肿瘤转移的指标,是预后不良的一组独立危险因素;DTC与突变型p53蛋白表达存在正相关。

Objective：We established nude mouse model of laryngocarcinoma and detect the disseminated tumor cell of peripheral blood and mutant p53 in tumour tissue, to research the correlation of DTC with variant p53 gene and their value of predicting cancer metastasis and prognosis in laryngocarcinoma. Methods. There are 40 nude mice in the experimental group which accepted hypodermic injection of laryngocarcinoma cell, 10 in control. By the reverse transcription polymerase chain reaction and nested polymerase chain reaction （Nest-PCR）,we tested CK19 mRNA in nude mouse＇s peripheral blood,as a marker of disseminated cancer cell at the 10th weeks after tumor-planting. We also tested the expression of p53 gene in tumour tissue by RT-PCR and immunohistology. After the nude dead, we dissect lung, liver and lymph node and use continuous pathological dyeing confirmed whether the transfer. Results：The positive rate of DTC was 50.0%（ 20/40 ）, but negative in the control group, the difference was significant（P〈0.05 ）. In tumor tissue, the positive rate of mutant p53 gene was 55.0% （ 22/40 ） ; The expression of DTC was positive correlation with p53. The mean one-year the mean survival time of DTC and p53 positive group was 34 weeks vs 43 weeks in negative group, the difference was significant （P〈0.05）. The positive of group have distant metastasis （38.9%）,but in negative group only（ 12.5% ）, the difference was significant （P〈0.05）. Gonclusion：The expression of DTC associated with variant p53 protein could be used as an indicator of tumor metastasis,and it might be an independent risk factor for an unfavourable prognosis ; DTC was positive correlation with variant p53 protein.