贝伐单抗联合培美曲塞对非小细胞肺癌移植瘤裸鼠体内IL-6、IL-17、IL-33及TNF-α的影响

Effect of Bevacizumab combined with Pemetrexed on expressions of IL-6, IL-17, IL-33 and TNF-α in non-small cell lung cancer xenografts of nude mice

目的通过建立非小细胞肺癌(NSCLC)移植瘤裸鼠模型,观察贝伐单抗联合培美曲塞对NSCLC移植瘤裸鼠肿瘤组织中白细胞介素(IL)-6、IL-17、IL-33及肿瘤坏死因子-α(TNF-α)表达水平的影响,进而探讨其可能机制。方法选择BALB/C雄性裸鼠60只,鼠龄3~4周,体质量13~14 g。按随机数字表法分为3组,即空白对照组、对照组及治疗组,每组20只。制备A549单细胞悬液,接种于对照组及治疗组裸鼠右侧近腋部皮下,空白对照组用0.9%氯化钠溶液(生理盐水)代替单细胞悬液。裸鼠全部成瘤后,治疗组裸鼠腹腔注射给药贝伐单抗和培美曲塞,空白对照组及对照组于同一时间采用等量0.9%氯化钠溶液代替。接种4周后,断颈处死裸鼠,采用酶联免疫吸附分析(ELISA)法和实时荧光定量聚合酶链反应(RT-PCR)法检测3组小鼠肿瘤组织中IL-6 mRNA、IL-17 mRNA、IL-33 mRNA及TNF-αmRNA和其蛋白的表达。结果对照组及治疗组40只裸鼠100%成瘤,且全部存活,移植瘤体积差异无统计学意义(P>0.05);与对照组相比,给药后,治疗组移植瘤体积生长开始受抑制,且在接种后27 d显著受到抑制(t=-34.42,P<0.05)。与空白对照组比较,对照组肿瘤组织中IL-6、IL-17、IL-33及TNF-α的mRNA和蛋白表达水平显著升高(P<0.05);经过贝伐单抗联合培美曲塞治疗后,与对照组比较,治疗组IL-6、IL-17、IL-33及TNF-α的表达mRNA和蛋白水平显著降低(P<0.05)。结论贝伐单抗联合培美曲塞可显著降低NSCLC移植瘤裸鼠肿瘤组织中IL-6、IL-17、IL-33及TNF-α的表达水平。

Objective To explore the molecular mechanism and effects of Bevacizumab combined with Pemetrexed on the expressions of interleukin（IL）-6, IL-17, IL-33 and tumor necrosis factor-α（TNF-α） in non-small cell lung cancer（NSCLC） model of nude mice. Methods Sixty BALB/C nude mice aged 3-4 weeks with body weight of 13-14 g were randomly divided into 3 groups（n = 20）： blank control group, control group and treatment group. The A549 cells were injected into right side of axillary subcutaneous of BALB/C nude mice in control group and treatment group, and blank control group was given the same equivalent 0.9 % Na Cl. After NSCLC model was successfully, the nude mice of treatment group were treated with Bevacizumab combined with Pemetrexed, the blank control group and control group were given the same equivalent 0.9 % Na Cl. Nude mice were sacrificed after A549 cells injection 4-week, then mRNA and protein expressions of IL-6, IL-17, IL-33 and TNF-α in3 groups were detected by enzyme linked immunosorbent assay（ELISA） and quantitative real-time PCR（RT-PCR） methods.Results Forty nude mice xenografts of control group and treatment group were successfully established and survived, the tu-mor volume between control group and treatment group was no significant difference（ P〉0.05）; Compared with control group,the drug inhibited tumor volume in treatment group, and was significantly inhibited at 27-day（ t =-34.42, P〈0.05）. Compared with blank control group, the mRNA and protein expression levels of IL-6, IL-17, IL-33 and TNF-α were significantly increased in control group（P〈0.05）. Compared with control group, after treatment of Bevacizumab combined with Pemetrexed,the mRNA and protein expression levels of IL-6, IL-17, IL-33 and TNF-α were significantly decreased in treatment group（P〈0.05）. Conclusion It is demonstrated that Bevacizumab combined with Pemetrexed could significantly decrease the expression levels of IL-6, IL-17, IL-33 and TNF-α in NSCLC xenografts of nude mice.