青藤碱对肺癌模型大鼠抑癌基因P16和P53的影响  

Effects of Sinomenine on Tumor Suppressor Gene P16 And P53 in Rats with Lung Cancer

作　　者：郑化军 王贤和 柯昌斌[2] 李莉[3] 

机构地区：[1]湖北医药学院附属人民医院,十堰442000 [2]十堰市太和医院,十堰442000 [3]湖北医药学院,十堰442000

出　　处：《世界科学技术-中医药现代化》2017年第10期1702-1706,共5页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：湖北省卫生厅青年人才攻关项目(WJ2015Q040):骨癌痛大鼠痛觉调控核团特导性基因与蛋白表达谱研究;负责人:柯昌斌

摘　　要：目的:本研究旨在探讨青藤碱对肺癌模型大鼠抑癌基因P16和P53的影响机制。方法:雄性SD大鼠40只,均采用左肺注射WALKER-256癌细胞悬液方法建立大鼠肺癌移植性模型,3周后筛选成瘤的大鼠30只,随机分为模型组、环磷酰胺组和青藤碱治疗组,另取健康SD大鼠10只设为正常对照组。青藤碱治疗组背皮下注射10%青藤碱治疗10周,环磷酰胺组注射环磷酰胺作为阳性对照,同时正常对照组和模型组注射生理盐水。治疗10周后取各组肺肿瘤测量瘤体积、瘤质量并计算抑瘤率;取肿瘤组织匀浆,流式细胞仪检测瘤体组织WALKER-256癌细胞在G1、G2、M和S四周期细胞比例;采用免疫组化法检测P16和P53蛋白阳性表达率,逆转录聚合酶链反应法(RT-PCR)测定P16mRNA和P53mRNA表达。结果:与模型组比较,青藤碱组抑瘤率达30.15%,P16和P53蛋白阳性表达率明显降低,P16mRNA和P53mRNA低表达,S期细胞比值、瘤体积和瘤质量明显降低、G1和G2期细胞比值提高(P<0.05)。结论:青藤碱可能通过影响抑癌基因P16和P53表达,调节G1和G2和S期细胞比值而起到抑制大鼠WALKER-256移植性肺癌肿瘤细胞的分化和增殖。This study was aimed to investigate the effect of sinomenine on the expression of tumor suppressor gene P16 and P53 in rats with lung cancer.A total of 40 male SD rats were treated by left-lung vein injection of WALKER-256 cell suspension to establish transplanted lung cancer model.After 3 weeks,30 rats screened of tumor were randomly divided into the model group,cyclophosphamide（CP） group and the sinomenine treatment group.Another 10 healthy SD rats were set as the normal control group.Sinomenine treatment group was treated with the subcutaneous injection of 10%sinomenine hydrochloride for 10 weeks.CP was injected in the CP group as positive control.The same amount of normal saline was injected in the normal control group and the model control group.After 10 weeks of treatments,lung tumors of each group were removed to measure the tumor volume and weight.And the tumor inhibition rate was calculated.Then,flow cytometry was used to detect the proportion of WALKER-256 cells in tumor tissues in G1,G2,M and S around four cycles.Immunohistochemistry was adopted to detect positive expression rates of P16 and P53 protein.Reverse transcription polymerase chain reaction（RT-PCR） were used to detect expression of P16 mRNA and P53 mRNA.The results showed that compared with the model control group,the inhibition rate of sinomenine group was 30.15%;the positive expression rate of P16 mRNA and P53 mRNA protein were significantly decreased;expressions of P16 mRNA and P53 mRNA were lower;tumor volume and tumor weight in S period got down significantly.The rates of cells in G1 and G2 periods got higher（P0.05）.It was concluded that sinomenine may inhibit the differentiation and proliferation of WALKER-256 transplanted lung cancer cells in rats by regulating the expression of tumor suppressor gene P16 and P53,regulating the ratio of cells in G1,G2 and S periods.

关 键 词：青藤碱 P16基因 P53基因 移植性肺癌 细胞周期 抑瘤率 

分 类 号：R96[医药卫生—药理学]