甲状腺低分化癌裸鼠移植瘤模型的建立及化疗敏感性研究

发布时间：2018-04-15 06:25

本文选题：移植瘤 + 裸鼠　；参考：《中国协和医科大学》2010年博士论文

【摘要】： [背景]甲状腺低分化癌和未分化癌是头颈部肿瘤恶性程度最高的肿瘤,目前主要采用手术、放疗和化疗的综合治疗,由于病变发展迅速、远处转移比例高,总体治疗效果差,由于发病率较低,较难开展大样本的随机对照研究,目前尚缺乏统一的治疗规范和化疗方案。局部复发和远处转移是主要致死原因,新的化疗药物尤其是分子靶向药物的出现,使研究者对化疗在提高局部控制和降低远处转移两方面的作用冀以新的希望,国外多个Ⅰ／Ⅱ期临床研究正在进行,目前国内尚无同类研究。 [目的]建立甲状腺低分化和未分化癌的裸鼠移植瘤模型；评估脂质体阿霉素、索拉非尼治疗甲状腺低分化癌移植瘤的疗效；筛选对甲状腺低分化和未分化癌细胞敏感的化疗药物,为临床治疗提供参考。 [方法]选取甲状腺乳头状癌、低分化癌和未分化癌的手术标本进行裸鼠皮下接种,通过连续传代建立稳定的移植瘤模型；用脂质体阿霉素(liposome doxorubicine)和索拉非尼(sorafenib, BAY 43-9006, Nexavar,多吉美)治疗裸鼠甲状腺低分化癌移植瘤,观察肿瘤生长情况,评估两种药物的疗效；分别利用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐[3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]及三磷酸腺苷肿瘤药物敏感性检测(adenosine triphosphate-tumor chemosensitivity assay, ATP-TCA)的方法对甲状腺癌细胞进行体外药物敏感性检测,筛选敏感药物。 [结果]3例甲状腺低分化和未分化癌裸鼠移植瘤模型均成功建立并传代,其中1例低分化癌模型连续传代后生长稳定,成瘤率达100%,而且肿瘤生长及信号传导相关蛋白表达稳定,适合进一步的动物实验；3例甲状腺乳头状癌移植瘤均未生长。利用脂质体阿霉素和索拉非尼对甲状腺低分化癌模型进行化疗,效果较明显,空白对照组、溶剂对照组、单药脂阿组、索拉非尼组、低量联合组、中量联合组、高量联合组化疗结束后移植瘤的最终平均体积分别为1274.13±393.76mm3、1060.00±469.05mm3、726.76±488.22mm3、451.54±97.75mm3、518.37±164.44mm3、310.51±210.53mm3和228.44±129.21mm3,瘤重分别为1.13±0.42g、0.91±0.39g、0.78±0.45g、0.55±0.17g、0.52±0.19g、0.34±0.21g和0.19±0.09g,单药脂阿组、索拉非尼组、低量联合组、中量联合组、高量联合组的抑瘤率分别为30.8%、40.8%、42.3%、62.9%和72.6%,高量联合组抑瘤率除与中量联合组无差异外(P=0.357)均高于其余各组,中量联合组优于单药脂阿组(P=0.001),而与索拉非尼组无差异(P=0.192)。各治疗组平均瘤重均明显低于空白对照组(P0.05)。各治疗组均无实验鼠死亡,高量联合组实验鼠体重在治疗过程中较其余各治疗组有明显减轻(P0.05)。利用MTT法发现索拉非尼对甲状腺乳头状癌、低分化和未分化癌均不同程度的敏感,而ATP-TCA方法则显示对3例甲状腺低分化和未分化癌均敏感的药物是长春新碱(VCR)、吉西他滨(GEM)以及顺铂+紫杉醇(DDP+PTX)的联合方案。 [结论]甲状腺低分化癌和未分化癌较乳头状癌移植成瘤率高,移植瘤模型经连续传代后稳定,适合动物药效学等研究；脂质体阿霉素和索拉非尼无论单药还是联合应用对本例甲状腺低分化癌移植瘤模型均有明显的抑瘤作用,中量联合疗效明显且副作用小。甲状腺低分化和未分化癌细胞药物敏感性个体差异较大,临床治疗前建议进行体外细胞药敏检测进行个体化的选择,ATP-TCA可能是一种比较理想的检测方法。
[Abstract]:background of the invention thyroid low differentiation cancer and undifferentiated carcinoma are the most malignant tumors in the head and neck , and the present invention mainly adopts the comprehensive treatment of surgery , radiotherapy and chemotherapy , and has the advantages of rapid lesion development , high distant metastasis ratio and poor overall treatment effect .

Objective To establish a model of nude mice transplantation tumor with low thyroid and undifferentiated carcinoma .
To evaluate the efficacy of liposomal doxorubicin and sorbiani in the treatment of low - differentiated thyroid carcinoma ;
Screening of chemotherapeutics sensitive to low thyroid and undifferentiated cancer cells provides a reference for clinical treatment .

Methods : The operation specimens of thyroid papillary carcinoma , low differentiation carcinoma and undifferentiated carcinoma were subcutaneously inoculated in nude mice , and stable transplanted tumor model was established by continuous passage .
To evaluate the effects of liposome doxorubescens and soralea , BAY 43 - 9006 , Nexavar , Dogius in the treatment of low - differentiated thyroid carcinoma in nude mice and to observe the growth of tumor and evaluate the efficacy of the two drugs .
The sensitivity of 3 - ( 4,5 - dimethylthiazol - 2 ) -2,5 - diphenyltetrazol - 3 - ( 4,5 - dimethylimidazol - 2 - yl ) -2,5 - diphenyltetrazol bromide ( MTT ) and adenosine triphosphate ( ATP - TCA ) was used to detect the drug sensitivity of thyroid cancer cells in vitro and to screen sensitive drugs .

Results Three cases of low differentiation and undifferentiated carcinoma of nude mice were successfully established and passaged . Among them , 1 case of low differentiation cancer model was stable after continuous passage , the rate of tumor formation reached 100 % , and tumor growth and signal transduction related protein expression was stable , which was suitable for further animal experiments ;
There was no difference between the two groups ( P = 0 . 05 ) . The tumor weight was significantly lower than that in the control group ( P = 0 . 05 ) . The tumor weight was 1 . 13 卤 393.76 mm3 , 0 . 78 卤 0 . 21 g , 0 . 91 卤 0 . 19 g , 0 . 34 卤 0 . 21 g and 0 . 19 卤 0 . 09 g .

Conclusion Low differentiation and undifferentiated carcinoma of the thyroid gland have higher tumor rate than papillary carcinoma , and the transplanted tumor model is stable after continuous passage and is suitable for animal pharmacodynamics and other studies .
In the present case , the tumor model of low differentiation and undifferentiated carcinoma of the thyroid has obvious tumor inhibiting effect , and the combined effect of the liposome doxorubicin and sorbiani is obvious and the side effect is small . The sensitivity of the thyroid low differentiation and the undifferentiated cancer cell is small . The sensitivity of the thyroid low differentiation and the undifferentiated cancer cell is large , and the individual selection of the in vitro cell drug sensitivity detection is recommended before clinical treatment , and the ATP - TCA may be a comparatively ideal detection method .

【学位授予单位】：中国协和医科大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R-332;R736.1

【参考文献】