T24-多柔比星原位膀胱癌多药耐药模型的建立及检测

 目的　建立T24-多柔比星（ADM）模拟人类原位膀胱癌多药耐药模型并行耐药性检测。方法　采用细胞移植法将T24-ADM耐药株及T24细胞敏感株移植到两组裸鼠膀胱后定期观察其一般情况。28 d后处死裸鼠，称成瘤膀胱质量、测体积，行组织病理学和细胞学检测，反转录聚合酶链反应（RT-PCR）检测多药耐药基因-1（MDR-1）的表达，MTT法测瘤组织制成的细胞悬液耐药性。结果　T24-ADM组及T24组裸鼠膀胱的成瘤率分别为80 %（8／10）和90 %（9／10），成瘤膀胱平均体质量和体积分别为 （0.8±0.3）g、（1.0±0.5）g、（875±158）mm3、（903±192）mm3，组间比较差异均无统计学意义（t＝1.332和t＝1.215，P＞0.05）。组织病理学：T24-ADM组4只出现右肾增大，T24组2只出现右肾增大，1.0 cm×1.8 cm，色暗灰到暗红。膀胱肿瘤组织切片细胞结构排列较乱，形态不规则，向肌层不同深度浸润，未突破浆膜层，两组间差异不明显。RT-PCR测MDR-1表达两组间差异有统计学意义（t＝3.612，P＜0.01）。MTT检测T24-ADM组裸鼠肿瘤细胞较T24组对ADM有明显的耐药性，差异有统计学意义（F＝412.107，P＜0.01），且对其他几种药物也有耐药性，差异有统计学意义（P＜0.05）。结论　细胞移植法成功建立了T24-ADM原位膀胱癌耐药模型，并具有多药耐药特性。

Construction and detection of multidrug resistance model in T24-ADM orthotopic bladder cancer

Objective To establish the orthotopic bladder cancer model of multidrug resistance as the human' s, and detect its resistance condition. Methods Two groups of nude rats 4-6 weeks of age were inculated with 1×107 cell of T24 or T24-ADM, following with observation and putting down their meat, drink,mental condition, urine and abdominal mass growth. Animals were sacrificed after 4 weeks later, then their bladder were weighted and measured, histopathologic assessment was performed,mdr1 was detected by PCR,and cells from the bladder tumors were detected of multidrug resistence by MTT. Results Group of nude rats inculated with T24-ADM generated tumors about 80 % (8/10), the one inculated with T24 was 90 % (9/10)and about 2-3 days early. The blank group had no rats emerge tumors in bladder mucosa at all. Bladder weight and volume: (0.8±0.3) g, (1.0±0.5) g, (875±158) mm3, (903±192) mm3, difference between the two groups had no significant (t = 1.332 and t = 1.215, P＞0.05). Histopathologic detection: The two groups of bladder cancer tissue biopsies can be seen more chaotic arrangement of cell structure, cell body shape is irregular, to the depth of myometrial invasion in different without breaking the film. Between the two groups there were no significantly differences. PCR detection of mdr1 expression differences between the two groups was significant (t = 3.612, P ＜0.01). Cytological detection of drug-resistant cell volume is slightly larger, and no significant difference in morphology. MTT detection: cells from the inculated T24-ADM mice bladder tumor were more resistance to ADM than the ones from the inculated T24 mice bladder tumor (F = 412.107, P＜0.01), and for several other drugs were also resistant. Conclusion Cell transplantation was successfully used to establish bladder cancer model in situ of T24-ADM, and with multi-drug resistance characteristics. The model laid the foundation for further multi-drug resistance research of bladder cancer.