Cheng Ming-rong, Li Qing, Hong Xiao-wu, Ye Tao, He Bing, Cheng Zhi-jian, Xu Hong-zhi, Zha Bing-bing
Department of General Sugery, Fudan University, Shanghai, China.
Zhonghua Zhong Liu Za Zhi. 2011 Sep;33(9):661-5.
To construct a recombinant plasmid pIRES-GM-CSF-IL-21, and to investigate its antitumor effect on tumors in the mice.
Fifty Bal b/c mice were included in this study. Cultured hepatoma H22 cells were inoculated in the left lobe of the liver to develop orthotopically transplanted liver tumor models. The mice with orthotopically transplanted liver tumor were randomly divided into 5 groups (n = 10): (1) Each mouse received injection of recombinant plasmid pIRES-GM-CSF-IL-21; (2) Received injection of plasmid pIRES-GM-CSF; (3) pIRES-IL-21; (4) Received injection of ampty plasmid pIRES (H22/neo group); (5) Received injection of PBS (H22 group) via the tail vein, respectively. Therefore, the anti-tumor effect was induced by both GM-CSF and IL-21, or by either of them alone. The serum levels of IFN-γ and IL-2 were detected by ELISA, and the cytotoxicity of spleen NK and CTL cells were tested by MTT colorimetry.
Comparing with the H22 and H22/Neo groups, the tumor weight in the mice of H22/GM-CSF group was (0.603 ± 0.223) g, H22/IL21-treated group (0.583 ± 0.290) g and H22/GM-CSF-IL21-treated group (0.303 ± 0.323) g, significantly lower than that in the H22 group [(1.591 ± 0.280) g] and H22/Neo group [(1.489 ± 0.155) g]. Among them the tumor growth was most significantly inhibited in the H22/GM-CSF-IL-21 group (0.303 ± 0.323) g, compared with that of H22 and H22/neo groups (P < 0.01). But there was no significant difference between the tumor weights of the H22/GM-CSF group and H22/IL-21 group, and between the tumor weights of the H22 and H22/Neo groups (P > 0.05). The levels of IFN-γ and IL-2 in peripheral blood of mouse models treated with H22/GM-CSF-IL-21 were significantly increased than that in the H22/GM-CSF group and H22/IL-21 group (all P < 0.01), but significantly decreased in the H22group and H22/Neo group (P < 0.01). The anti-tumor activity of splenic NK cells and CTLs in the H22/GM-CSF-IL21 group was significantly enhanced (P < 0.01), compared with the significantly decreased in the H22 and H22/Neo groups.
Our results demonstrate apparent antitumor effect of the plasmid pIRES-GM-CSF-IL-21 on the orthotopically transplanted liver tumor in mice. The combination of both pIRES-GM-CSF and IL-21 is more effective than that of pIRES/IL21 or pIRES/GM-CSF treatment alone. In addition, the plasmid pIRES-GM-CSF-IL-21 can also promote the secretion of IFN-γ and IL-2 in vivo, and enhance the cytotoxic activity of splenic NK and CTLs against the transplanted liver tumor.
构建重组质粒pIRES-GM-CSF-IL-21，并研究其对小鼠肿瘤的抗肿瘤作用。
本研究纳入50只Bal b/c小鼠。将培养的肝癌H22细胞接种于肝脏左叶，建立原位移植性肝肿瘤模型。将原位移植性肝肿瘤小鼠随机分为5组（n = 10）：（1）每只小鼠接受重组质粒pIRES-GM-CSF-IL-21注射；（2）接受质粒pIRES-GM-CSF注射；（3）pIRES-IL-21；（4）接受空质粒pIRES注射（H22/neo组）；（5）经尾静脉接受PBS注射（H22组）。因此，GM-CSF和IL-21联合或单独均可诱导抗肿瘤作用。采用ELISA法检测血清IFN-γ和IL-2水平，采用MTT比色法检测脾脏NK和CTL细胞的细胞毒性。
与H22组和H2（2/Neo组相比，H22/GM-CSF组小鼠肿瘤重量为（0.603±0.223）g，H22/IL21治疗组为（0.583±0.290）g，H22/GM-CSF-IL21治疗组为（0.303±0.323）g，显著低于H22组[（1.591±0.280）g]和H22/Neo组[（1.489±0.155）g]。其中，H22/GM-CSF-IL-21组肿瘤生长抑制最为显著（0.303±0.323）g，与H22组和H22/neo组相比（P<0.01）。但H22/GM-CSF组与H22/IL-21组肿瘤重量之间，以及H22组与H22/Neo组肿瘤重量之间无显著差异（P>0.05）。H22/GM-CSF-IL-21处理的小鼠模型外周血中IFN-γ和IL-2水平显著高于H22/GM-CSF组和H22/IL-21组（均P<0.01），但在H22组和H22/Neo组中显著降低（P<0.01）。与H22组和H22/Neo组显著降低相比，H22/GM-CSF-IL21组脾脏NK细胞和CTLs的抗肿瘤活性显著增强（P<0.01）。
我们的结果表明质粒pIRES-GM-CSF-IL-21对小鼠原位移植性肝肿瘤具有明显的抗肿瘤作用。pIRES-GM-CSF和IL-21联合使用比单独使用pIRES/IL21或pIRES/GM-CSF治疗更有效。此外，质粒pIRES-GM-CSF-IL-21还可促进体内IFN-γ和IL-2的分泌，并增强脾脏NK和CTLs对移植性肝肿瘤的细胞毒性活性。
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