索拉非尼联合贝伐单抗对小鼠宫颈癌细胞株U14移植瘤抑制作用的机制研究  

Study on the mechanism of Sorafenib combined with Bevacizumab on the tumor inhibition of cervical cancer cell line U14 transplantation in mice

作　　者：刘志英[1] 袁淑芳[2] Liu Zhiying;Yuan Shufang(Department of Pathology, Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830001, China;Department of Infection, Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830001, China)

机构地区：[1]新疆医科大学第五附属医院病理科,乌鲁木齐830001 [2]新疆医科大学第五附属医院感染科,乌鲁木齐830001

出　　处：《国际外科学杂志》2019年第5期315-320,F0004,共7页International Journal of Surgery

基　　金：新疆维吾尔自治区自然科学基金(2015211C171).

摘　　要：目的通过索拉非尼联合贝伐单抗对小鼠宫颈癌细胞株U14的作用,观察细胞株U14的移植瘤细胞结构的变化及对Bcl2、Bax蛋白表达的调节作用。方法健康近交系雌性615小鼠60只,4~6周龄;平均体重20.4 g,范围18~25 g。选取10只小鼠作为空白对照组(A组),另外通过接种小鼠宫颈癌细胞株U14建立荷瘤小鼠模型,在皮下接种U14的第6天,挑取肿瘤直径≥5 mm的荷瘤鼠40只,按照完全随机化分组的方法将小鼠分为5组,分别为模型组(B组)、索拉非尼组(C组)、低剂量贝伐单抗联合索拉非尼组(D组)、中剂量贝伐单抗联合索拉非尼组(E组)及高剂量贝伐单抗联合索拉非尼组(F组),每组各8只。治疗方案为:A组及B组小鼠均给予1 ml 0.9%氯化钠溶液,C组小鼠给予索拉非尼12 mg/kg,D组小鼠给予贝伐单抗2.5 mg/kg+索拉非尼12 mg/kg,E组小鼠给予贝伐单抗5 mg/kg+索拉非尼12 mg/kg,F组小鼠给予贝伐单抗10 mg/kg+索拉非尼12 mg/kg。均采用腹腔注射,用药24 d后,处死小鼠。每6 d用天平称瘤重(g),观察各组小鼠的体重变化,并计算各组小鼠的抑瘤率;通过苏木精-伊红染色光镜下观察各组小鼠的移植瘤组织病理变化;通过电镜观察各组小鼠的移植瘤组织细胞形态变化;通过免疫组化法光镜下观察各组小鼠的Bcl-2及Bax蛋白表达情况。计量资料以均数±标准差(Mean±SD)表示,组间比较采用单因素方差分析,两两比较采用LSD法。结果在药物腹腔注射开始至最后1 d,小鼠体重开始缓慢持续增加,各组小鼠趋势基本一致。C组小鼠抑瘤率为8.02%,D组为4.92%,E组为11.10%,F组为5.42%。E组抑瘤率最高,效果相对最好。A组与其余各组瘤重质量比较,差异具有统计学意义(P<0.05)。电镜结果显示,C、D、E、F组细胞结构变化相似,均在处理后出现了肿瘤细胞的凋亡,E组凋亡情况最好;在C、D、E、F组凋亡相关蛋白Bax均出现表达上调,E组阳性数量最多;C、D、E、F组原癌Objective According to effect of Sorafenib combined Bevacizumab in mice U14 cervical cancer cell lines, observe the changes of cell lines of transplanted tumor U14 cell structure and adjustment effect of Bcl2 and Bax protein expression. Methods Sixty female 615 mice of healthy inbreeding line, aged from 4 to 6 weeks, average weight is 20.4 g, range from 18 to 25 g. The animals were divided into blank control group (group A)with 10 mice, and U14 cervical cancer cell lines by vaccinating mice a tumorburdened mice model was established, on the 6th day after subcutaneous inoculation of U14, 40 tumorburdened mice with tumor diameter ≥ 5 mm were selected. The mice were divided into 5 groups according to the completely randomized grouping method: model group (group B), Sorafenib group (group C), lowdose Bevacizumab combined with Sorafenib group (group D), the middle dose Bevacizumab combined with Sorafenib group (group E), and the high dose Bevacizumab combined with Sorafenib group (group F), 8 mice in each group. The treatment regimen consisted of 1 ml of 0.9% sodium chloride solution in group A and group B, Sorafenib 12 mg/kg in group C, and Bevacizumab 2.5 mg/kg + Sorafenib 12 mg/kg in group D, Bevacizumab 5 mg/kg+Sorafenib 12 mg/kg in group E, and Bevacizumab 10 mg/kg + Sorafenib 12 mg/kg in group F. All mice were injected intraperitoneally and sacrificed by dislocation 24 days later. The tumor weight (g) was measured every 6 days, and the body weight of each group was observed, and calculated the tumor inhibition rate of each group of mice. The histopathological changes of the transplanted mice were observed by hematoxylin-eosin staining. The transplantation of each group of mice was observed by electron microscope. Morphological changes of tumor tissue;the expression of Bcl-2 and Bax protein in each group were observed by immunohistochemistry. The measurement data were expressed as mean standard deviation(Mean±SD), univariate analysis of variance was used for inter-group comparison, and LSD method w

关 键 词：宫颈肿瘤 抗肿瘤联合化疗方案 药物疗法 联合 基因  bcl-2 BAX蛋白 小鼠 

分 类 号：R737.33[医药卫生—肿瘤]