60^Co源照射诱导肺癌细胞凋亡及其凋亡小体体外负载自体及同种异体树突状细胞的实验研究

Experimental study of lung cancer apoptosis induced by ^(60)Co irradiation and its apoptotic body loaded with autologous and allogeneic dendritic cells in vitro

目的通过放射线诱导肺癌细胞凋亡小体,并使其在体外负载自体及同种异体树突状细胞(DC),为诱导特异性肿瘤细胞免疫治疗提供大量抗原提呈细胞。方法手术切除肺癌组织并分离培养肺癌细胞;利用免疫组化法鉴定肺癌细胞;利用60Coγ射线照射肺癌细胞诱导凋亡小体;通过流式细胞术(FCM)、DNA琼脂糖凝胶电泳(直接上样法)法检测细胞凋亡;应用共同孵育法使自体及同种异体DC体外负载肺癌细胞凋亡小体,并进行扫描电镜观察。结果通过组织分离法获得单个肿瘤细胞,经60Coγ射线照射可诱导大量凋亡小体,和自体及同种异体DC共孵育后,凋亡小体被DC吞噬或包裹。结论经60Coγ射线照射诱导的凋亡小体可作为抗原负载自体及异体DC用于诱发特异性抗肿瘤免疫反应,为肿瘤的主动免疫治疗提供了新的途径。

Objective Through inducing lung cancer apoptosis by radiation exposure method and its apoptotic body loaded with autologous and allogeneic dendritic cell （DC） in vitro, to provide a great quantity antigen presenting cells （APCs） for inducing specific antitumor immune response. Methods Lung cancer tissue were cut by operation to isolate and culture lung cancer cells. Immunohistochemistry method was used to identify lung cancer cell. Apoptotic body was induced by lung cancer cells irradiated by 60^Coγ ray. Apoptosis was determined by flow cytometry and DNA agarose gel electrophoresis （ direct loaded method）. Autologous and allogeneic DCs were loaded with lung cancer cell apoptotic body in vitro by co-incubation method and observed by scanning electron microscope. Results Single tumor cells obtained by tissue separation method induced a lot of apoptotic body through 60^Coγ ray irradiation. After co-incubation with autologous and allogeneic DC, apoptotic bodies were phagocytized and packaged by DCs. Conclusions Apoptotic body induced by 60^Coγ ray irradiation can be used as antigen loaded with autologous and allogeneic DC to induce specific antitumor immune response. This experiment provides a new way for tumor active immunotherapy.