摘 要 目的: 观察重组改构人肿瘤坏死因子（rmhTNF-α）协同顺铂（cisplatin ,DDP）对小鼠Lewis肺癌移植瘤的抑制作用，并探讨其可能的作用机制。方法：将Lewis肺癌细胞接种于C57BL/6小鼠右腋皮下，随机分为4组：生理盐水组(对照组)、DDP组（3 mg/kg）、rmhTNF-α组（150×104 U/kg）、rmhTNF-α（150×104 U/kg）+DDP（3 mg/kg）组。于细胞接种的第7天，肿瘤直径约0.6 cm时，瘤内注射给药，连续3 d，停药1 d后处死小鼠，剥离并称取瘤重，计算抑瘤率。流式细胞术测定移植瘤细胞凋亡率及细胞周期， RT-PCR检测移植瘤组织Survivin的表达。结果：（1）rmhTNF-α+DDP组治疗的抑瘤率（36.61%）明显高于DDP组（17.12%）或rmhTNF-α组（15.83%）单药治疗（P<0.05）。两药联合使用的q=1.2，具有良好的协同作用。（2）联合用药治疗后移植瘤细胞的凋亡率［（28.2±1.8）%］显著高于DDP（21.6±1.0）%和rmhTNF-α［（19.3±2.0）%单药治疗(P<0.05)；前者的细胞周期明显阻滞于G2期。（3）3个治疗组移植瘤细胞Survivin 基因的表达受到明显抑制（P<0.01）,联合用药组的抑制程度较单药组更明显（P<0.05）。结论：rmhTNF-α与DDP联合应用具有协同抗Lewis肺癌移植瘤的作用，其机制可能与抑制Survivin基因表达、诱导肿瘤细胞凋亡有关。

Abstract Objective: To investigate the inhibitory effect of recombinant mutant human tumor necrosis factor (rmhTNF-α) combined with cisplatin (DDP) against transplanted Lewis lung carcinoma (LLC) in mice, and to explore the related mechanism. Methods: LLC cells were subcutaneously inoculated into C57BL/6 mice at the right axilla and the mice were randomly divided into 4 groups: sodium chloride (control group),DDP group, rmhTNFα group, rmhTNFα plus DDP group. On the 7th day after inoculation the diameter of the tumor reached 0.6 cm; the corresponding agents were intratumorally injected for 3 d. All mice were sacrificed 1 d later and the tumors were obtained, weighed, and the tumor inhibitory rate was calculated. Flow cytometric analysis was used to examine cells apoptosis and cell cycle. Expression of survivin mRNA was examined by RTPCR.Results: (1)The tumor inhibitory rates of the DDP plus rmhTNFα group (3661%) was significantly higher than those of the DDP group (17.12%) and rmhTNF-α group (15.83%, P＜0.05）.The q value of Jin′s formula was 1.2, indicating a good synergistic effect between rmhTNFα and DDP. (2) The cell apoptotic rate of the rmhTNFα plus DDP group (\[28.2±1.8\]%) was significantly higher than those of the DDP group (\[21.6±1.0\]%) and rmhTNFα group (\[19.3±2.0\]%, P＜0.05）; the cells were arrested G2 phase in the DDP plus rmhTNFα group. (3) RTPCR showed that the expression of survivin gene was significantly inhibited in all the 3 groups, with the inhibition in the DDP plus rmhTNFα group more potent than that in the other 2 groups (P＜0.01). Conclusion: The combination of rmhTNFα and DDP has synergic effect in treatment of transplanted Lewis lung cancer, which might be related to the inhibition of survivin gene expression and induction of tumor cell apoptosis.

河北省科技攻关计划（No.072761147)