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目的探讨肿瘤相关糖蛋白72(TAG72)靶向性嵌合锚定T淋巴细胞的制备方法,并检测它对TAG72阳性肝癌细胞增殖的阻滞效应。方法分离外周血单核细胞(PBMC),然后用免疫磁珠法分离得到CD8+T淋巴细胞。将重组真核表达载体anti-TAG72-scFv-CD28-pcDNA3.0采用脂质体介导的细胞转染和细胞培养,以制备TAG72靶向性的嵌合锚定T淋巴细胞;将嵌合锚定T淋巴细胞与TAG72阳性肝癌细胞SMMC7721共培养,通过流式细胞仪检测肝癌细胞的周期变化,分析嵌合锚定T淋巴细胞对肝癌细胞增殖的抑制效应。结果 TAG72靶向性嵌合锚定T淋巴细胞可识别肝癌细胞SMMC7721;用流式细胞仪检测发现,嵌合锚定T淋巴细胞可引起肝癌细胞SMMC7721的增殖阻滞。结论 TAG72靶向性嵌合锚定T淋巴细胞可特异性识别TAG72阳性肝癌细胞SMMC7721并引起其增殖阻滞。
Objective To investigate the preparation method of tumor-associated glycoprotein 72 (TAG72) targeting chimeric anchored T lymphocytes and to detect its blocking effect on the proliferation of TAG72 positive hepatoma cells. Methods Peripheral blood mononuclear cells (PBMCs) were isolated and then CD8 + T lymphocytes were isolated by immunomagnetic beads method. The recombinant eukaryotic expression vector anti-TAG72-scFv-CD28-pcDNA3.0 was transfected into cells by liposome-mediated transfection and cell culture to prepare TAG72-targeting chimeric anchored T lymphocytes. T lymphocytes and TAG72 positive hepatocellular carcinoma cells SMMC7721 were co-cultured, the cycle changes of hepatoma cells were detected by flow cytometry, and the inhibitory effect of chimeric anchored T lymphocytes on the proliferation of hepatoma cells was analyzed. Results TAG72 targeting chimeric anchored T lymphocytes could identify hepatocellular carcinoma cells SMMC7721. Using flow cytometry, it was found that chimeric anchored T lymphocytes could induce the proliferation arrest of hepatocellular carcinoma cells SMMC7721. Conclusion TAG72 targeting chimeric anchored T lymphocytes can specifically recognize TMC72 positive hepatocellular carcinoma cells SMMC7721 and cause its proliferation arrest.