Stimulation of DC-CIK with PADI4 Protein Can Significantly Elevate the Therapeutic Efficiency in Esophageal Cancer
Stimulation of DC-CIK with PADI4 Protein Can Significantly Elevate the Therapeutic Efficiency in Esophageal Cancer
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PADI4蛋白刺激DC-CIK可显着提高食管癌的治疗效率
DOI：
10.1155/2019/6587570
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发表时间：
2019-01-01
影响因子：
4.1
通讯作者：
Chang, Xiaotian
中科院分区：
文献类型：
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作者：
Liu, Chunyan;Zheng, Yingying;Chang, Xiaotian
PADI4在许多肿瘤中广泛表达。本研究应用PADI4作为肿瘤标志物刺激DC-（树突状细胞-）CIK（细胞因子诱导杀手），这是一种免疫治疗方法。方法用PADI4表达质粒转染ec源性ECA-109细胞。将PADI4基因插入原核表达载体中，得到重组蛋白。用PADI4过表达细胞的裂解液或纯化的重组PADI4蛋白来装载dc，然后将细胞与CIK细胞共孵育。流式细胞术检测DC和CIK细胞表型。台盼蓝染色分析CIK细胞的增殖和活力。采用CCK8检测DC-CIK细胞对培养的ECA-109细胞的细胞毒作用。通过注射ECA-109细胞制备荷瘤小鼠。将PADI4过表达细胞或PADI4重组蛋白的裂解液刺激DC-CIK细胞注射到荷瘤小鼠体内。采用磁共振成像（MRI）检测肿瘤生长情况。结果与过表达padi4的细胞裂解液孵育后，CD40+ dc的比例增加了17.5%。padi4刺激的dc诱导CIK细胞增殖率提高53.2%，CIK细胞杀伤ECA-109细胞的能力提高12.1%。用padi4过表达细胞的裂解物刺激DC-CIK细胞，使荷瘤小鼠的肿瘤体积减少18.6%。重组PADI4蛋白对CIK细胞的增殖和细胞毒性的影响与PADI4过表达细胞的裂解物相似。此外，重组蛋白使CD40+ DCs的比例提高了111.8%，CD80+ DCs提高了6.3%，CD83+ DCs提高了30.8%，CD86+ DCs提高了7.8%。在动物模型中，用rpadi4刺激的dc诱导CIK细胞可使细胞增殖提高50.3%，CIK细胞杀伤ECA-109细胞的能力提高14.7%，肿瘤体积降低35.1%。结论PADI4刺激DC-CIK细胞，通过促进DC成熟、CIK细胞增殖和细胞毒性，显著抑制荷瘤小鼠的肿瘤生长。PADI4可能是一种潜在的肿瘤标志物，可用于提高DC-CIK细胞的治疗效率。
Background PADI4 has extensive expression in many tumors. This study applied PADI4 as a tumor marker to stimulate DC- (dendritic cell-) CIK (cytokine-induced killer), an immunotherapy approach. Methods A PADI4 expression plasmid was transfected into EC-originating ECA-109 cells. PADI4 gene was also inserted into a prokaryotic expression vector to produce recombinant protein. Lysate from PADI4-overexpressing cells or the purified recombinant PADI4 protein was used to load DCs, and the cells were then coincubated with CIK cells. DC and CIK cell phenotypes were determined using flow cytometry. The proliferation and viability of CIK cells were analyzed using trypan blue staining. The cytotoxic effect of DC-CIK cells on cultured ECA-109 cells was determined using CCK8 assays. Tumor-bearing mice were prepared by injection of ECA-109 cells. DC-CIK cells stimulated with lysate from PADI4-overexpressing cells or the PADI4 recombinant protein were injected into the tumor-bearing mice. The tumor growth was measured with magnetic resonance imaging (MRI). Results Following incubation with lysate from PADI4-overexpressing cells, the ratio of CD40+ DCs increased by 17.5%. Induction of CIK cells with PADI4-stimulated DCs elevated the cell proliferation by 53.2% and the ability of CIK cells to kill ECA-109 cells by 12.1%. DC-CIK cells stimulated with lysate from PADI4-overexpressing cells suppressed tumor volume by 18.6% in the tumor-bearing mice. The recombinant PADI4 protein showed a similar effect on CIK cell proliferation and cytotoxicity as that of the lysate from PADI4-overexpressing cells. Furthermore, the recombinant protein elevated the ratio of CD40+ DCs by 111.8%, CD80+ DCs by 6.3%, CD83+ DCs by 30.8%, and CD86+ DCs by 7.8%. Induction of CIK cells with rPADI4-stimulated DCs elevated the cell proliferation by 50.3% and the ability of CIK cells to kill ECA-109 cells by 14.7% and suppressed tumor volume by 35.1% in the animal model. Conclusion This study demonstrates that stimulation of DC-CIK cells with PADI4 significantly suppressed tumor growth in tumor-bearing mice by promoting DC maturation, CIK cell proliferation, and cytotoxicity. PADI4 may be a potential tumor marker that could be used to improve the therapeutic efficiency of DC-CIK cells.