从胆囊癌的一种新血供方式——血管生成拟态看胆囊癌诊疗观点改变

Insight into diagnosis and treatment of gallbladder cancer from vasculogenic mimicry:a new blood supply for gallbladder cancer

从胆囊癌的一种新的血液供应方式——血管生成拟态(vasculogenic mimicry,VM)论述胆囊癌诊疗观点的改变。1994.1—2000.12同济医院确诊的74例原发性胆囊癌标本及其临床病理学和人口统计学资料;H&E染色+CD31/PAS双重染色(Envision法),胆囊癌VM(+)/(-)临床参数单/多因素、Cox模型风险和Kaplan-Meier生存分析+log-rank检验。细胞、动物实验:胆囊癌GBC-SD、SGC-996细胞二/三维培养,侵袭力、迁移力、VM形成测定和倒置光显微镜、透射/扫描电镜VM形态学观察以及动态磁共振荷瘤鼠胆囊癌移植瘤血流动力学检测。信号通道研究:胆囊癌细胞三维培养和裸鼠移植瘤VM组织切片的免疫组化、免疫荧光、Western blotting、QRT-PCR检测,以探讨相关信号通道。13.5%(10/74)胆囊癌存VM,其中4.3%(13/306)管腔内含红细胞,即胆囊癌VM是高倍光学显微镜下由胆囊癌细胞围成管腔、内衬癌细胞而非血管内皮细胞、有圈PAS阳性界膜分隔、腔内含红细胞的管道结构;VM(+)胆囊癌腺癌居多,有较高的血行转移机会;VM是影响胆囊癌患者预后的独立危险因素,有VM存在胆囊癌患者生存期短,预后差。体外胆囊癌GBC-SD细胞三维培养、荷瘤鼠胆囊癌石蜡切片经H&E染色和CD31/PAS双重染色,光/电镜观察,85.7%存在VM,其VM结构与临床结果一致;MRI增强扫描提示,荷瘤裸鼠胆囊癌移植瘤中心或瘤体周围存在VM血供而不致出现肿瘤坏死而呈现MRI强化信号。发现胆囊癌存在VM-相关PI3-K/MMPs/Ln-5γ2和Eph A2/FAK/Paxillin信号通路。高侵袭胆囊癌存在VM这一特殊生物学特性。因高侵袭胆囊癌VM不能被常规病理学检测,故建议对临床恶性程度高、易血行转移、预后差胆囊癌,常规开展胆囊癌标本CD31/PAS双重染色检查。对胆囊癌根治术后复发、辅助化疗+抗血管生成靶向疗效差者,应考虑存在VM,采取针对性的"抗血管生成拟态"辅助治疗。

From the discovery of a new blood supply of gallbladder carcinoma：vasculogenic mimicry（VM）, insight into diagnosis and treatment of gallbladder cancer. 74 cases of primary gallbladder carcinoma diagnosed at Tongji Hospital from 1994.1 to 2000.12 and their clinical pathological and demographic data were used;VM of gallbladder carcinoma was detected by H＆E staining and CD31-PAS double staining（Envision method）,single-or multiple-factor clinical parameters,Cox model,Kaplan-Meier survival analysis and log-rank test for gallbladder carcinoma with VM （＋）/（-） were done. Two or three-dimensional cultures of GBC-SD and SGC-996 cells,invasion,migration,VM formation assay and hemodynamic of gallbladder cancer in vitro and in vivo were carried out. VM-related signal pathways were studied by immunohisto- chemistry, immunofluorescence,Western blotting and QRT-PCR. VM in 13.5%（10/74） of gallbladder cancers was found,namely VM of gallbladder cancer is the pipeline structures surrounded by gallbladder cancer cells and non vascular endothelial cells,with the PAS positive,red blood cells inside under the microscope;VM（＋）gallbladder cancers being mainly adenocarcinoma with highly blood metastasis;VM is an independent prognostic factor for gallbladder cancer patients with VM,and prognosis of gallbladder cancer patients with VM is poor. VM was found in GBC-SD cell 3-D matrices and xenografts,and the formation of VM through the activation of the PI3K/MMPs/Ln-5γ2 signaling pathway. There is a special biological characteristic of VM in highly aggressive gallbladder carcinoma;because VM cannot be detected by conventional pathology,CD31-PAS double staining for specimens were routinely done in the gallbladder cancer patient with highly malignant,easy to hematogenous metastasis and poor prognosis;the presence of VM should be considered for post-operational recurrence after gallbladder cancer radical resection,poor results of adjuvant chemotherapy and anti-angiogenic targeted therapy,＂anti vasculogenic mimicry＂adjuvant therapy was adopted.