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目的制备肿瘤相关糖蛋白-72抗原(TAG-72)嵌合锚定T细胞,通过体外实验检测其对乳腺癌细胞的抑癌活性。方法分离健康人外周血单个核细胞(PBMC),采用脂质体lipofectamineTM2000介导的细胞瞬时转染方法,将已制备的重组真核表达载体即抗TAG-72-scFv-CD3ζ-pcDNA 3.0导入PBMC,以获得TAG-72嵌合锚定T细胞,以此为转染组;用转染空载体pcDNA 3.0的PBMC作为对照组。将转染组嵌合锚定T细胞和对照组PBMC细胞分别与乳腺癌细胞系MCF-7及Bcap37共培养,观察两者对乳腺癌细胞G1期的阻滞率。结果转染组对MCF-7细胞系的G1期阻滞率为(82.3±6.9)%,对照组为(43.4±3.9)%,2组间比较差异具有统计学意义(P<0.05);转染组对Bcap37细胞系的G1期阻滞率为(51.3±4.7)%,对照组为(45.6±2.5)%,2组间比较差异无统计学意义(P>0.05)。结论 TAG-72嵌合锚定T细胞可特异性地抑制TAG-72+乳腺癌细胞的增殖,此将为乳腺癌的临床诊治提供一种有希望的途径。
Objective To prepare tumor-associated glycoprotein-72 antigen (TAG-72) chimeric and anchored T cells and test its anti-tumor activity against breast cancer cells in vitro. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteers. The recombinant eukaryotic expression vector, anti-TAG-72-scFv-CD3ζ-pcDNA 3.0, was introduced into PBMC by lipofectamineTM2000 mediated transient transfection. , To obtain TAG-72 chimeric anchored T cells as a transfected group; PBMC transfected with empty vector pcDNA 3.0 as a control group. The transfected group of chimeric anchored T cells and control PBMCs were co-cultured with breast cancer cell lines MCF-7 and Bcap37 to observe the G1 arrest rate of breast cancer cells. Results The G1 arrest of MCF-7 cell line was (82.3 ± 6.9)% in the transfected group and (43.4 ± 3.9)% in the control group, with statistical significance (P <0.05) The G1 arrest of Bcap37 cell line was (51.3 ± 4.7)% in the control group and (45.6 ± 2.5)% in the control group, but there was no significant difference between the two groups (P> 0.05). Conclusion TAG-72 chimeric anchored T cells can specifically inhibit the proliferation of TAG-72 + breast cancer cells, which will provide a promising approach for the clinical diagnosis and treatment of breast cancer.