Kim Tae-Jung, Moon Hyong Woo, Kang Sungmin, Yang Jonghyup, Hong Sung-Hoo, Lee Ji Youl, Ha U-Syn
Department of Hospital Pathology, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
Department of Urology, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
J Cancer. 2019 Jun 2;10(14):3259-3266. doi: 10.7150/jca.30079. eCollection 2019.
: To explore whether cultured CTC from bladder-cancer patients originate from bladder cancer and share chromosomal abnormalities, by means of a fluorescence hybridization (FISH) test. : A total of 15 ml of blood was collected from the patients with bladder cancer before treatment began. Isolated CTCs were divided into 5 ml for CTC enumeration and 10 ml for CTC culture. CTCs were counted by immunofluorescent staining with vimentin, cytokeratin, CD45, and DAPI antibody. CTCs were cultured using isolated CTCs in 96-well plates of Mesenchymal Stem Cell Growth Medium for 16~18 days. The resulting cultured CTCs from 20 men with bladder cancer were analyzed by Urovysion FISH. : Common gains were on chromosome 3, 7, and 17 in 20 (74.1%), 14 (51.9%), and 20 (74.1%) of 27 patients, respectively. Polysomy was detected on chromosomes 3 and 7 in 9 patients (33.3%). Polysomy involving two chromosomes was observed in 16 (59.3%, chromosome 3 and 17) and 9 patients (33.3%, chromosome 7 and 17) in the same cell. Among the patients with isolated gain, 17 (63.0%) met the positive criteria for Urovysion FISH. Homozygous deletion of 9p21, 5 (18.5%) involved more than 12 cells. Among the different patient cohorts, positive results based on the Urovysion criteria were obtained in cultured CTCs derived from 19 (70.4%) patients. : Application of FISH Urovysion to cultured CTCs from bladder cancer could be an effective first step to confirm their origin and sharing of chromosomal abnormalities.
为了通过荧光原位杂交(FISH)试验探究膀胱癌患者培养的循环肿瘤细胞(CTC)是否源自膀胱癌并存在染色体异常。收集15例膀胱癌患者治疗前的血液。分离出的CTC分为5ml用于CTC计数,10ml用于CTC培养。通过波形蛋白、细胞角蛋白、CD45和DAPI抗体的免疫荧光染色对CTC进行计数。将分离出的CTC在间充质干细胞生长培养基的96孔板中培养16至18天。对20例男性膀胱癌患者培养得到的CTC进行Urovysion FISH分析。27例患者中,分别有20例(74.1%)、14例(51.9%)和20例(74.1%)在染色体3、7和17上出现常见增益。9例患者(33.3%)在染色体3和7上检测到多体性。在同一细胞中,观察到16例(59.3%,染色体3和17)和9例患者(33.3%,染色体7和17)存在涉及两条染色体的多体性。在孤立增益的患者中,17例(63.0%)符合Urovysion FISH的阳性标准。9p21纯合缺失,5例(18.5%)涉及超过12个细胞。在不同患者队列中,源自19例(70.4%)患者的培养CTC根据Urovysion标准获得阳性结果。将FISH Urovysion应用于膀胱癌培养的CTC可能是确认其来源和染色体异常共享的有效第一步。