摘要
目的探讨自制的靶向IL18-EGF融合蛋白抑制人肝癌细胞株SMMC-7721增殖的作用及对人肝癌裸鼠动物模型抑瘤率的影响,为人肝癌的生物治疗提供新思路。方法注射SMMC-7721细胞至Balb/c裸鼠右侧腰部,成瘤后,将裸鼠进行随机分成IL18-EGF组、rhIL18组和NS对照组,瘤周注射相应的药物及其经X-射线处理的NK-92细胞。在治疗前后各时间点计量肿瘤体积,最后一次治疗的次日,处死全部裸鼠,切除肿瘤组织。一部分肿瘤组织制成单细胞悬液并应用流式细胞术进行细胞周期和凋亡的监测;另一部分肿瘤组织用于病理切片染色和观察。结果在治疗结束时,移植瘤的体积在IL18-EGF组、rhIL18组和NS组分别为(0.063±0.030)cm3,(0.212±0.037)cm3和(0.432±0.030)cm3,NS组与IL18-EGF组和rhIL18组有显著性差异(P<0.01),rhIL18组与IL18-EGF组有显著性差异(P<0.05);细胞周期分析表明,IL18-EGF组、rhIL18组和NS组分别有(96.11%±1.51%),(81.3%±2.02%)和(72.18%±0.46%)的细胞停滞在G1期,IL18-EGF与后两者相比差异均有显著性(P<0.05,P<0.01),rhIL18组与NS组相比差异也有显著性(P<0.05),S期细胞的比例分别为(3.69%±0.41%),(16.51%±1.25%)和(23.85%±1.36%),IL18-EGF与后两者相比差异有显著性(P<0.01,P<0.01),rhIL18组与NS组相比也有明显差异(P<0.05);肿瘤细胞自主凋亡发生率在3组中也差异有显著性,与对照组(7.10%±1.13%)相比,IL18-EGF组[(28.05%±3.89%),P<0.05]、rhIL18组[(25.48%±5.46%),P<0.05]肿瘤细胞自主凋亡差异有显著性,而IL18-EGF组和rhIL18组间差异无显著性;肿瘤标本切片观察未发现各组治疗前后病理特征有明显变化。结论裸鼠瘤周注射IL18-EGF合并NK细胞能抑制SMMC-7721移植瘤的生长表明IL18-EGF具有体内的抗肿瘤效应,可能为人肝癌治疗提供一种新的治疗策略与手段。
[Objective] To investigate the inhibition proliferation effect of the targeting IL18-EGF fused protein created by ourselves on human hepatic carcinoma cell lines SMMC-7721 cells in vivo and the inhibitory rate in nude mice model so as to seeking a new way for biological therapy of human hepatic carcinoma. [Methods] SMMC-7721 celts were subcutaneously injected into the right flank of Balb/e nude mice, The mice with established SMMC-7721 xenograft were randomized into three groups of treatment： IL18-EGF, rhlL18 and NS blank group. Tu- mor-beating mice were treated with peritumoral subcutaneously injection of NK-92 cells irradiated with X-ray fol- lowed by peritumoral subcutaneously injection of the three groups. The next morning after the last dosage, all mice were sacrificed and the tumor masses and mouse organs were completely dissected for further usage. [Results] Before the end of therapy, we found volume of xenograft tumor was （0.063±0.030） mm3 in IL18-EGF group, （0.212±0.037） mm3 in rhIL18 group, and （0.432±0.030） mm3 in control group respectively. Compared to control, SMMC- 7721 xenograft growth in Balb/c nude mice was markedly inhibited with peritumoral injection of IL18-EGF or rhIL18 （P 〈0.01）. Furthermore, the inhibitory effect of IL18-EGF on SMMC-7721 growth in vivo was more profound than that of rhIL18 （P 〈0.05）. Meanwhile, more tumor ceils were arrested in G1 phase in IL18-EGF group （96.11%±1.51%）, compared to rhIL18 group [（81.3%±2.02%）, P 〈0.05] as well as to control [（72.18%±0.46%）, P 〈0.01]. The difference on the percentage of G1 phase between rhIL18 group and control was also significant （P 〈0.05）. The pereentages of S phase in IL18-EGF group, rhlL18 group and control were （3.69%±0.41%）, （16.51%±1.25%）, and （23.85%±1.36%）, respeetively （IL18-EGF vs rhIL18： P 〈0.01; IL18-EGF vs control： P 〈0.01; IL18 vs eontrol： P 〈 0.05）. Tumor spontaneous apoptotie rates in IL18-EGF, rhIL18 and control group were （28.05%±3.89%） （P 〈0.05, eompared to control）, （25.48%±5.46%） （P 〈0.05, compared to control） and （7.10%±1.13%）, respeetively. And there was still not signifieant difference between IL18-EGF and rhIL18 group on spontaneous apoptosis induction. No sig- nificant difference was observed on pathologic features of SMMC-7721 xenograft grown in Balb/e nude mice treated with IL18-EGF, rhlL18 and control, respectively. [Conclusions] Subcutaneous injeetion of IL18-EGF and NK eells adjacent to tumor significantly inhibited the growth of SMMC-7721 transplantation tumor whieh suggested that ILl 8-EGF fused protein owned the effeet of the antineoplastie activity in vivo and can be an effeetive means in the biologieal therapy.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2009年第6期805-810,共6页
China Journal of Modern Medicine
基金
温州市科技项目(Y2006A001)
