SNU-16胃癌原位模型建立和药物A药效验证毕业论文
2022-06-04 22:58:02
论文总字数:14905字
摘 要
动物模型在新药筛选中起到了重要作用,本研究尝试通过SNU-16-Luc细胞株建立良好的胃癌原位的动物模型,评估出紫杉醇对胃癌的药效。首先,在体外扩增SNU-16-Luc胃癌细胞至对数生长期,收集细胞并按1×107 细胞量/只动物皮下异位接种在BALB/c裸鼠上,定期观测肿瘤生长,并测定肿瘤活体荧光情况来判定肿瘤细胞的活力情况,待肿瘤体积生长至300-500mm3时,进行瘤块转接种至BALB/c裸鼠的胃大弯处,之后定期通过活体成像来确定肿瘤生长情况,接种后7天后,进行荧光测定,并根据荧光测定的结果进行随机分组进行药效实验,每组6只动物,第一组:给生理盐水,尾静脉注射(i.v.), 一周两次给药(biw), 给药3周时间;第二给:给紫杉醇,尾静脉注射(i.v.),一周两次给药(biw),给药3周时间.每天观察并记录动物的状态,每周2次动物的体重,给后药第0天、第7天、第14天、第21天进和荧光成像,根据数据形成小鼠体重变化曲线、荧光增长曲线,以确定药效情况。实验在给药后21天结束,结束取小鼠的胃和肿瘤,并称重,拍照;并剥离下肿瘤后,称瘤重,拍照。并把取得的肿瘤进行速冻和固定,用于后期的蛋白表达分析和病理分析。根据瘤重数据,计算肿瘤抑制率。药效学研究表明,受试药物紫衫醇能抑制小鼠体内SNU-16-Luc肿瘤的生长,同时对小鼠无明显毒副作用,表明受试药物紫杉醇对该类型肿瘤有一定的治疗效果。
关键词: SNU-16-Luc 胃癌 BALB/c裸鼠 活体成像
Abstract
Animal model plays an important role in new drug screening, this study try to through the SNU - 16 - Luc good subcutaneous inoculation of gastric cancer cell lines to establish animal model of evaluation of paclitaxel on the efficacy of gastric cancer. First of all, in vitro amplification SNU - 16 - Luc gastric cancer cells to the logarithmic phase, collecting cells and according to the quantity of 1 x 107 cells/animal subcutaneous ectopic vaccination in BALB/c nude mice, regular observation of tumor growth, and determination of tumor fluorescent living conditions to determine the activity of tumor cells was, for tumor volume growth to 300-500 mm3, turn to carry on the plaque vaccination to BALB/c nude mouse stomach big bend, after tumor growth by in vivo imaging on a regular basis to determine the situation, in 7 days after inoculation, fluorescence determination, and according to the fluorescence determination results of randomized efficacy experiments, each group of six animals, the first group, physiological saline, caudal vein injection (i.v.), twice a week (biw), for 3 weeks; To 2: Z to drugs, tail intravenous injection (i.v.), twice a week (biw), for three weeks. A day to observe and record the state of the animal, the animal's weight, 2 times a week to 0, 7 days, after 14 days, 21 days and fluorescence imaging, according to the data form curves of body weight in mice, fluorescent growth curve, to determine the efficacy. Experiment at the end of the 21 days after the treatment, end of stomach and tumor in mice, and weighing, taking pictures; And stripping off after tumor, tumor weight, taking pictures. And the tumors were frozen and fixed, for later analysis of the protein expression and the pathological analysis. According to the tumor weight data, calculate the tumor inhibition rate. Pharmacodynamic studies have shown that a drug to purple shirts alcohol can inhibit SNU - 16 - Luc tumor growth in mice, and no obvious side effects in mice, show that the tested drug taxol on this type of tumor has a certain therapeutic effect.
Keywords: SNU - 16 - Luc ;bare stomach ;BALB/c mice ;in vivo imaging
目录
摘要 I
Abstract II
第一章 文献综述 1
1.1 前言 1
1.2 文献综述 1
第二章 实验材料与方法 4
2.1 实验材料 4
2.1.1 细胞株及细胞准备 4
2.1.2 实验动物 4
2.1.3 试剂和药物 4
2.1.4 主要仪器及设备 5
2.2 实验方法 5
2.2.1 细胞培养方法 5
2.2.2 细胞接种方法 6
2.2.3 瘤块转接种 7
2.2.4 动物观察与测量 8
2.2.5分组与给药 8
2.2.6 荧光测量 9
2.2.7 结束实验及取材 9
2.2.8 数据分析及统计 9
第三章 实验结果与讨论 10
3.1 受试药物对肿瘤细胞在小鼠胃部生长的影响 10
3.2 受试药物对于荷瘤小鼠肿瘤重量的影响 12
3.3 受试药物对于荷瘤小鼠体重的影响 13
3.4 荷瘤小鼠生存及死亡情况 14
3.5 结论 14
第四章 展望 14
参考文献 15
致谢 17
第一章 文献综述
1.1 前言
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