环磷酰胺暴露后Piwil2在人神经母细胞瘤移植瘤中的表达  

Expression of Piwil2 in xenograft tissue with human neuroblastoma after exposure to cyclophosphamide

作　　者：王城[1,2] 何文飞[1,2] 张红梅[3] 赵丹[1,2] 杨金凤[1,2] 廖君左[1,2] 周柯均[1,2] 

机构地区：[1]川北医学院附属医院儿外科国家卫生部内镜诊疗技术培训基地 [2]川北医学院肝胆胰肠疾病研究所,四川南充637000 [3]川北医学院附属医院肿瘤科,四川南充637000

出　　处：《中国生物制品学杂志》2014年第11期1412-1415,共4页Chinese Journal of Biologicals

基　　金：四川省教育厅基金资助项目(13ZB0245;12ZA050)

摘　　要：目的探讨环磷酰胺（cyclophosphamide,CP）暴露后Piwil2在人神经母细胞瘤（neuroblastoma,NB）移植瘤中的表达。方法建立免疫活性C57BL/6j系小鼠的人NB SK-N-SH细胞株荷瘤模型,选择瘤体积在100~200 mm3之间的20只荷瘤小鼠,随机分为生理盐水对照组（NS组）和环磷酰胺组（CP组）,每组10只,CP组每日经腹腔注射CP75 mg/kg,2次/周,连续2周;NS组每日经腹腔注射NS 2 ml。每2天测量肿瘤体积,计算相对肿瘤体积（RTV）;停药后次日处死小鼠,称瘤重,计算抑瘤率（IR）;并采用RT-PCR法检测各组移植瘤组织中Piwil2基因m RNA的转录水平,免疫组化法和Western blot法检测各组移植瘤组织中Piwil2蛋白的分布及表达水平。结果与NS组比较,CP组移植瘤的RTV和瘤重均显著降低（P〈0.05）,CP组的平均抑瘤率为83.56%。两组移植瘤组织中Piwil2基因m RNA的转录水平差异无统计学意义（P〉0.05）。NS组移植瘤组织中Piwil2蛋白高表达于胞浆,而CP组大部分肿瘤细胞坏死,Piwil2蛋白表达下降,细胞核稀疏,核浓缩,细胞体积变小;CP组移植瘤组织中Piwil2蛋白的表达水平明显低于NS组（P〈0.05）。结论 CP干预后,移植瘤缩小,移植瘤组织中Piwil2基因m RNA的转录水平不变,但蛋白表达水平有改变,可能影响人NB抵抗性。Objective To analyze the expression of Piwil2 in xenograft with neuroblastoma（NB） after exposure to cyclophosphamide（CP）. Methods The 3-week-old immunocompetent C57 BL / 6j mice were transplanted with SK-N-SH cells. Twenty mice bearing tumors at sizes of 100 ~ 200 mm3 were divided into two groups randomly,i.e. physiological saline（NS）control and CP groups,ten for each. The mice in CP group were injected i.p. with CP at a dosage of 75 mg / kg,twice a week for 2 weeks,while those in NS group with 2 ml of NS. The tumor size was measured once 2 d,based on which the relative tumor volume（RTV） was calculated. The mice were killed on the second day after withdrawing of administration,of which the tumors were weighed,and the tumor inhibiting rate（IR） was calculated. The transcription levels of Piwil2 m RNA in the transplanted tumors in two groups were determined by RT-PCR,while the expression level of Piwil2 protein by immunohistochemical assay and Western blot. Results The RTV and tumor weight in CP group decreased significantly as compared with those in NS group（P 〈 0. 05）,while the mean tumor inhibiting rate was 83. 56%.The transcription levels of Piwil2 m RNAs in two groups showed no significant difference（P 〉 0. 05）. The expression level of Piwil2 protein in xenograft tissue was significantly higher than that in cytoplasm in NS group. However,necrosis of most of tumor tissue was observed in CP group,while the expression level of Piwil2 protein decreased,the nuclei were sparse in distribution and concentrated,and the cell volume was small. However,The expression level of Piwil2 proteinin xenograft tissue of CP group was significantly lower than that of NS group（P 〈 0. 05）. Conclusion After treatment with CP,the xenograft was shrunk,and the transcription level of Piwil2 m RNA showed no significant change,while the protein expression level changed significantly,which might be associated with the resistance induced by neuroblastoma.

关 键 词：Piwil2 神经母细胞瘤 移植瘤 环磷酰胺 

分 类 号：R739.42[医药卫生—肿瘤]