[1]杨永君,李瑶,尹立,等.抗间皮素嵌合抗原受体修饰T细胞治疗卵巢癌的实验研究[J].第三军医大学学报,2018,40(11):945-953.
 YANG Yongjun,LI Yao,YIN Li,et al.Anti-mesothelin chimeric antigen receptor modified T cells for ovarian cancer[J].J Third Mil Med Univ,2018,40(11):945-953.
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抗间皮素嵌合抗原受体修饰T细胞治疗卵巢癌的实验研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第11期
页码:
945-953
栏目:
基础医学
出版日期:
2018-06-15

文章信息/Info

Title:
Anti-mesothelin chimeric antigen receptor modified T cells for ovarian cancer
作者:
杨永君李瑶尹立刘新东卞修武余时沧
陆军军医大学(第三军医大学)第一附属医院:全军临床病理学研究所,乳腺外科
Author(s):
YANG Yongjun LI Yao YIN Li LIU Xindong BIAN Xiuwu YU Shicang

Institute of Pathology, Department of Breast Surgery, First Affiliated Hospital, Army Military Medical University (Third Military Medical University), Chongqing, 400038, China

关键词:
卵巢肿瘤间皮素嵌合抗原受体修饰T细胞杀伤作用
Keywords:
ovarian cancer mesothelin chimeric antigen receptor modified T cells killing effect
分类号:
R394.3; R73-362; R737.31
文献标志码:
A
摘要:

目的     观察抗间皮素(mesothelin,MSLN)嵌合抗原受体修饰的T细胞对卵巢癌细胞的杀伤作用,探索新的卵巢癌治疗手段与策略。方法     设计人源化抗人间皮素的嵌合抗原受体融合基因;构建慢病毒表达载体;感染人外周血单核细胞,制备稳定表达抗MSLN嵌合抗原受体的T细胞(chimeric antigen receptormodified T cells, CART);Western blot、qRT-PCR、流式细胞术检测CAR的表达;免疫组织化学法和Western blot检测卵巢癌样本及细胞系中MSLN表达;体内和体外实验验证CART细胞对卵巢癌细胞的杀伤能力。结果     与正常组织相比,卵巢癌组织MSLN表达显著升高(P<0.01);成功制备了抗MSLN-CART细胞;体外实验证明,抗MSLN-CART细胞能够特异性识别和杀伤MSLN阳性的卵巢癌细胞,效靶比为2-4∶1时,MSLN-CART杀伤率仍达到80%;NOD/SCID小鼠卵巢癌原位移植瘤模型证明,经尾静脉回输抗MSLN-CART细胞(1×107/只,共2次),原位移植瘤消失。结论     抗MSLN-CART细胞能有效、特异且安全地杀伤卵巢癌细胞,抑制肿瘤的生长。

Abstract:

Objective    To observe the killing effect of anti-mesothelin (MSLN) chimeric antigen receptor (CAR) modified T cells (CART) for ovarian cancer cells, and explore new treatment methods and strategies for the tumor. Methods    A humanized CAR structure of anti-human MSLN was designed and ligated to a lentiviral vector. Human peripheral blood mononuclear cells were infected with associate lentivirus for introduction of CART. Western blotting, qRT-PCR and flow cytometry were used to detect the correct expression of CAR structure on the membrane surface of CART. The expression of MSLN in ovarian cancer samples and cell lines was detected by immunohistochemical assay and Western blotting. The cytotoxicity of the CART cells to kill MSLN-positive ovarian cancer cells was evaluated via in vitro and in vivo experiments. Results    Compared with normal tissues, ovarian cancer tissues highly expressed MSLN (P<0.01); The anti-MSLN-CART cells were successfully prepared, and these cells specifically recognized and killed MSLNpositive ovarian cancer cells in vitro experiments. When the E∶T ratio was 2-4∶1, the killing rate still remained at 80%. Moreover, the orthotopic xenografts of ovarian cancers in NOD/SCID mice disappeared completely after totally 1×107 MSLN-CART cells were injected via the tail vein twice. Conclusion    Anti-MSLN-CART cells can effectively, specifically and safely kill ovarian cancer cells, inhibit the growth of tumors.

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更新日期/Last Update: 2018-06-13