复方苦参注射液对奥沙利铂耐药直肠癌裸鼠移植瘤的影响及与NOX1通路的关系

Effects of Compound Kushen Injection on Oxaliplatin-Resistant SW1463 Rectal Cancer Tumor-bearing Nude Mice and NOX1 pathway

目的:观察复方苦参注射液(Compound Kushen Injection,CKI)对奥沙利铂耐药SW1463直肠癌裸鼠移植瘤的抑瘤作用,并进一步探讨其作用机制与NOX1信号通路的关系。方法:诱导SW1463直肠癌细胞对奥沙利铂耐药后,建立SW1463耐药裸鼠移植瘤模型,将40只荷瘤鼠随机分为对照组、CKI低剂量组[1.0 mL/(kg·d)]、CKI中剂量组[2.0 mL/(kg·d)]和CKI高剂量组[4.0 mL/(kg·d)],记录荷瘤体积,持续干预28 d后处死动物取材,ELISA法检测移植瘤中活性氧(ROS)含量水平,qPCR法检测移植瘤中NADPH氧化酶(NOX)1和NOXO1的mRNA转录水平,ELISA法检测移植瘤中NOX1和NOXO1蛋白表达水平。结果:不同干预剂量CKI均能够显著抑制SW1463耐药裸鼠移植瘤的生长,降低移植瘤质量分数(P<0.05 vs.对照组),并呈一定程度的剂量依赖性,其中CKI高剂量组抑瘤作用最佳(P<0.05 vs.CKI低、中剂量组);不同干预剂量CKI均能够显著抑制SW1463耐药裸鼠移植瘤中ROS生成(P<0.05 vs.对照组),其中CKI高剂量组ROS水平最低(P<0.05 vs.CKI低、中剂量组);不同干预剂量CKI均能够显著抑制SW1463耐药裸鼠移植瘤中NOX1和NOXO1的mRNA相对转录水平和蛋白表达水平(P<0.05 vs.对照组),其中CKI高剂量组抑制作用最强(P<0.05 vs.CKI低、中剂量组)。结论:复方苦参注射液能够显著抑制SW1463耐药裸鼠移植瘤的生长,其作用机制可能是通过NOX1信号通路调控ROS水平从而发挥抗奥沙利铂耐药直肠癌作用的。

Objective:To observe the anti-tumor effect of Compound Kushen Injection(CKI)on oxaliplatin-resistant SW1463 rectal cancer xenografts in nude mice,and further explore the relationship between its mechanism and NOX1 signaling pathway.Methods:After inducing the resistance of SW1463 cells to oxaliplatin,the xenograft model of SW1463 resistant nude mice was established.Forty tumor-bearing mice were randomly divided into control group,low-dose CKI group[1.0 mL/(kg·d)],medium-dose CKI group[2.0 mL/(kg·d)]and high-dose CKI group[4.0 mL/(kg·d)].The tumor-bearing volume was recorded and the animals were killed after 28 days of continuous intervention.The levels of ROS was detected by ELISA.NOX1 and NOXO1 transcriptions were detected by qPCR and NOXO1 protein expression was detected by ELISA.Results:CKI with different intervention doses could significantly inhibit the growth of transplanted tumors and reduce the mass fraction of transplanted tumors in SW1463 resistant nude mice(P<0.05 vs.control group)in a dose-dependent manner.CKI with high dose had the best anti-tumor effect(P<0.05 vs.CKI with low and medium dose).CKI with different doses could significantly inhibit ROS production in SW1463-resistant nude mice xenografts(P<0.05 vs.control group),with the lowest ROS level in CKI high dose group(P<0.05 vs.CKI low and medium dose group).CKI with different intervention doses could significantly inhibit the relative transcriptional and protein levels of NOX1 and NOXO1 in SW1463-resistant nude mice transplanted tumors(P<0.05 vs.control group),and CKI with high dose had the strongest inhibitory effect(P<0.05 vs.CKI with low and medium dose).Conclusion:CKI could significantly inhibit the growth of SW1463-resistant transplanted tumors in nude mice.Its mechanism may be that it regulates ROS level through NOX1 signaling pathway to play an anti-oxaliplatin-resistant role in colorectal cancer.