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目的探讨肿瘤抗原负载的DC与DC-CIK细胞对B16恶性黑色素瘤荷瘤鼠的治疗作用。方法在无菌条件下提取小鼠骨髓干细胞,应用相应的细胞因子诱导出DC与CIK细胞;然后通过肿瘤全细胞冻融法将DC进行肿瘤抗原负载;将抗原负载的DC与CIK细胞按1:10的比例进行共培养,即是肿瘤抗原负载的DC-CIK细胞。建立B16恶性黑色素瘤模型鼠。将30只B16恶性黑色素瘤模型鼠随机分为三组,分别注射肿瘤抗原负载的DC、DC-CIK细胞、生理盐水(对照组)。测量治疗前后肿瘤体积的变化,计算抑瘤率。通过HE染色切片观察比较在对照组和不同治疗组中肿瘤细胞数量及形态学变化;通过免疫组化染色法,检测MMP-3分别在不同组中的表达。结果治疗组均有明显的抗肿瘤作用,抗原负载的DC-CIK明显优于抗原负载DC的治疗作用;MMP-3的表达在抗原负载的DC-CIK治疗组明显低于抗原负载的DC治疗组,而对照组MMP-3则高表达。结论抗原负载的DC-CIK和DC有明显的抗肿瘤作用,DC-CIK优于DC,这一作用与其对MMP-3表达的调节有关。
Objective To investigate the therapeutic effect of tumor antigen loaded DCs and DC-CIK cells on B16 malignant melanoma bearing mice. Methods The mouse bone marrow stem cells were extracted under aseptic conditions. DCs and CIK cells were induced by corresponding cytokines. DCs were then loaded with tumor antigens by freeze-thaw method. Antigen-loaded DCs and CIK cells were treated with 1: 10 ratio of co-culture, that is, tumor antigen-loaded DC-CIK cells. Establishment of B16 malignant melanoma model mice. Thirty B16 malignant melanoma models were randomly divided into three groups: DCs, DC-CIK cells loaded with tumor antigens and normal saline (control group) were injected respectively. Measurement of tumor volume before and after treatment changes, calculate the inhibition rate. The numbers and morphological changes of tumor cells in the control group and different treatment groups were observed and compared by HE staining sections. The expression of MMP-3 in different groups was detected by immunohistochemical staining. Results The treatment group had obvious antitumor effect. The DC-CIK loaded with antigen was significantly better than that treated with antigen-loaded DC. The expression of MMP-3 in DC-CIK group was significantly lower than that in antigen-loaded DC group , While the control group was high expression of MMP-3. Conclusion Antigen-loaded DC-CIK and DC have obvious anti-tumor effect, and DC-CIK is better than DC, which is related to the regulation of MMP-3 expression.