CAJ | 학술논문

目的:携带癌胚抗原(carcinocmbryonic antigen,CEA)基因的重组人腺相关病毒(reconstructive human adeno-association virus,rh-AAV)感染树突状细胞(dendritic cell,DC)诱导获得抗原特异性细胞毒性T淋巴细胞(cytotoxic lymphocyte,CTL),体外检测其对CD44+CD24-/low乳腺癌干细胞的杀伤效果.方法:分离供者外周血单个核细胞,以细胞因子白介素-4(interleukin-4,IL-4)、粒细胞-巨噬细胞克隆刺激因子(granulocyte-macrophage colony-stimulating factor,GM-CSF)和肿瘤坏死因子α(tumor necrosis factor-alpha,TNF-α)诱导培养获得DC,细胞因子白介素-2(interleukin-2,IL-2),刺激培养获得T淋巴细胞.含CEA基因的rh-AAV感染培养DC,诱导成熟后将DC和T细胞混合培养获得CTL细胞.流式分选MCF-7和MA-MDB-231细胞系中CD44+CD24-/low乳腺癌干细胞,MTT法检测CTL细胞对CD44+CD24-/low乳腺癌干细胞的杀伤效果.结果:MCF-7和MDA-MB-231中CD44+/CD24-/low群比例分别为5.1%和76.3%.CEA基因转染DC诱导的CTL细胞对表达CEA抗原的MCF-7杀伤率为46.5%±15.0%,与未转染组相比,差异有统计学意义(P=0.009);对MCF-7细胞中分选的CD44+CD24-/low乳腺癌干细胞的杀伤率为44.7%±28.2%,明显高于未转染组.对非乳腺癌干细胞杀伤率为50.6%±22.2%,与未转染组相比,差异有统计学意义(P=0.05).在不表达CEA基因的MDA-MB-231乳腺癌细胞,CEA转染诱导的CTL细胞对未分选细胞、分选的CD44+/CD24-/low亚群和非干细胞亚群的杀伤率与未转染的对照组相比,差异均无统计学意义(P＞0.05).结论:携带CEA基因rh-AAV转染DC诱导产生的抗原特异性CTL细胞可杀伤表达CEA的乳腺癌细胞,对CD44+CD24-/low乳腺癌干细胞也具有一定的杀伤活性,提示免疫治疗可能是治疗乳腺癌干细胞潜在有效的手段.
목적:휴대암배항원(carcinocmbryonic antigen,CEA)기인적중조인선상관병독(reconstructive human adeno-association virus,rh-AAV)감염수돌상세포(dendritic cell,DC)유도획득항원특이성세포독성T림파세포(cytotoxic lymphocyte,CTL),체외검측기대CD44+CD24-/low유선암간세포적살상효과.방법:분리공자외주혈단개핵세포,이세포인자백개소-4(interleukin-4,IL-4)、립세포-거서세포극륭자격인자(granulocyte-macrophage colony-stimulating factor,GM-CSF)화종류배사인자α(tumor necrosis factor-alpha,TNF-α)유도배양획득DC,세포인자백개소-2(interleukin-2,IL-2),자격배양획득T림파세포.함CEA기인적rh-AAV감염배양DC,유도성숙후장DC화T세포혼합배양획득CTL세포.류식분선MCF-7화MA-MDB-231세포계중CD44+CD24-/low유선암간세포,MTT법검측CTL세포대CD44+CD24-/low유선암간세포적살상효과.결과:MCF-7화MDA-MB-231중CD44+/CD24-/low군비례분별위5.1%화76.3%.CEA기인전염DC유도적CTL세포대표체CEA항원적MCF-7살상솔위46.5%±15.0%,여미전염조상비,차이유통계학의의(P=0.009);대MCF-7세포중분선적CD44+CD24-/low유선암간세포적살상솔위44.7%±28.2%,명현고우미전염조.대비유선암간세포살상솔위50.6%±22.2%,여미전염조상비,차이유통계학의의(P=0.05).재불표체CEA기인적MDA-MB-231유선암세포,CEA전염유도적CTL세포대미분선세포、분선적CD44+/CD24-/low아군화비간세포아군적살상솔여미전염적대조조상비,차이균무통계학의의(P＞0.05).결론:휴대CEA기인rh-AAV전염DC유도산생적항원특이성CTL세포가살상표체CEA적유선암세포,대CD44+CD24-/low유선암간세포야구유일정적살상활성,제시면역치료가능시치료유선암간세포잠재유효적수단.