氟尿嘧啶注射液及氟尿嘧啶植入剂对食管癌细胞的抑制效果和临床应用研究

发布时间：2018-06-08 07:11

本文选题：5-氟尿嘧啶 + 氟尿嘧啶植入剂　；参考：《西南医科大学》2017年硕士论文

【摘要】：目的:食管癌是发病率较高的常见恶性肿瘤,在恶性肿瘤的死亡原因中排在前列,氟尿嘧啶作为消化道恶性肿瘤化疗的传统经典药物,在消化道恶性肿瘤的治疗中发挥重要作用。本实验为进一步探讨抗肿瘤药物5-氟尿嘧啶(5-Fu)类药物在食管癌治疗中的应用潜力,我们通过体外实验设计验证了5-氟尿嘧啶及氟尿嘧啶植入剂对人食管癌TE-1细胞株生长的杀伤作用和诱导凋亡的能力,探寻有效的诱导肿瘤细胞凋亡的浓度和作用时间,为临床制定合理的化疗方案提供实验室依据。通过临床应用研究进一步验证了氟尿嘧啶植入剂治疗食管癌的安全性及有效性。方法:第一部分:以体外培养的人食管癌TE-1细胞株为实验研究对象,分别用5-氟尿嘧啶和氟尿嘧啶植入剂作用于TE-1,分别在不同时间点进行细胞计数,找出凋亡明显的药物浓度及作用时间,并利用细胞凋亡检测试剂盒进一步检测不同质量浓度及不同时间5-氟尿嘧啶及氟尿嘧啶植入剂对TE-1细胞生长的影响,以期望筛选出最适合的药物质量浓度和作用时间。具体的实验设计包括以下两个方面,(1)5-氟尿嘧啶对TE-1细胞增殖的抑制作用:根据文献数据和预实验将5-氟尿嘧啶组设四个浓度组0μg/ml、10μg/ml、50μg/ml、100μg/ml,分别处理TE-1细胞24h、48h、72h、96h后进行细胞计数,找出凋亡明显的药物浓度及作用时间,并且利用细胞凋亡检测试剂盒进行凋亡检测。(2)氟尿嘧啶植入剂对TE-1细胞增殖的抑制作用:用氟尿嘧啶为内标物,利用高效液相色谱法(HPLC)检测,绘制氟尿嘧啶标准曲线,测量不同时间氟尿嘧啶植入剂的释放浓度,依次在24h、48h、72h、96h处理te-1细胞,找出凋亡明显的药物浓度及作用时间,并进行细胞的凋亡检测。第二部分:收集本院住院患者中行氟尿嘧啶植入剂治疗食管癌的病例,在内镜直视下,将病灶以1.0cm的层间距分为若干层,每层内以每间隔1.0cm种植一枚缓释化疗粒子;植入时每点植入10mg,依病情需要,每4周植入1次,种植3次为1个疗程。此项研究中全部病患均顺利完成治疗。通过比对研究病患在治疗前、治疗后3个月及治疗后6个月的临床症状缓解情况、病灶面积变化情况、吞咽困难改善情况、不良反应、生活质量等。结果:(1)5-氟尿嘧啶对人食管癌细胞株te-1细胞周期的抑制与药物处理的时间具有很强的相关性,在浓度为10μg/ml时对te-1作用明显,增加药物浓度并不能够明显地增加药物作用的效果,且作用96小时时开始产生明显差异,但进行凋亡检测未见明显的凋亡细胞。(2)使用高效液相色谱(hplc)测定氟尿嘧啶植入剂释放量,发现缓释化疗粒子释放的药物浓度随着时间的延长逐渐增加,到了48h以后基本维持平衡了,而且后面的浓度反而有点下降,氟尿嘧啶植入剂处理te-1细胞,发现不同时间有不同作用影响,当48h-72h时间段细胞数减少的最明显,作用浓度约为282μg/ml,96小时内作用时间越长作用越明显,并且能够明显地检测到凋亡细胞(3)治疗前分别与治疗3、6个月比较,患者病变面积、吞咽困难分级变化差异有统计学意义(p0.05);但生活质量改善情况差异无统计学意义(p0.05,原因可能是随访时间较短及样本量较少,未体现出个体之间差异);另外治疗前与治疗3个月白细胞(wbc)差异有统计学意义(p0.05);治疗前、治疗3和6个月并发症情况发生率差异无统计学意义。结论:1.氟尿嘧啶注射液及氟尿嘧啶植入剂对人食管癌细胞株均有抑制其增殖的作用。2.氟尿嘧啶植入剂释放浓度稳定,有能促进人食管癌细胞株TE-1凋亡的作用,且96小时内时间越长作用越明显。3.氟尿嘧啶植入剂对食管癌细胞株的抑制效果优于氟尿嘧啶注射液。4.内镜下植入5-氟尿嘧啶(5-FU)缓释粒子治疗晚期食管癌,是一种安全有效、毒副作用小、耐受性好的姑息治疗方法。
[Abstract]:Objective: esophageal cancer is a common malignant tumor with high incidence of malignant tumor, and it is the leading cause of the death of malignant tumors. Fluorouracil plays an important role in the treatment of digestive malignant tumors. This experiment is a further study of the antitumor drug 5- fluorouracil (5-Fu). In the treatment of esophageal cancer, we verified the killing and apoptosis ability of 5- fluorouracil and Fluorouracil Implants on the growth of human esophageal cancer TE-1 cell lines by experimental design in vitro, to explore the effective induction of the concentration and time of the apoptosis of tumor cells, and to provide an experiment for the clinical formulation of a rational chemotherapy scheme. The safety and effectiveness of Fluorouracil Implants for the treatment of esophageal cancer was further verified by clinical application. Methods: Part 1: the TE-1 cell line of human esophageal cancer cells cultured in vitro was used as the experimental object, and 5- fluorouracil and Fluorouracil Implants were used for TE-1 respectively, and the cell count at different time points was carried out respectively. To find out the obvious drug concentration and action time of apoptosis, and to further detect the effect of 5- fluorouracil and Fluorouracil Implants on the growth of TE-1 cells at different mass concentration and different time by cell apoptosis detection kit, in order to screen out the most suitable drug quality concentration and time. Specific experimental designs include The next two aspects, (1) the inhibitory effect of 5- fluorouracil on the proliferation of TE-1 cells: according to the literature data and pre experiment, four concentration groups of 5- fluorouracil group were set up 0 mu g/ml, 10 mu g/ml, 50 mu g/ml, 100 mu g/ml, respectively, to deal with TE-1 cell 24h, 48h, 72h, after 96h, the cell count was carried out to find apoptotic drug concentration and action time, and the use of cells, and the use of cells Apoptosis detection kit for apoptosis detection. (2) the inhibitory effect of Fluorouracil Implants on the proliferation of TE-1 cells: using fluorouracil as the internal standard, using high performance liquid chromatography (HPLC) detection, draw the standard curve of fluorouracil, measure the release concentration of Fluorouracil Implants at different time, and in sequence in 24h, 48h, 72h, 96h to treat TE-1 cells. The concentration and time of the apoptotic drugs and the time of action were detected and the cell apoptosis was detected. The second part: collect the cases of the treatment of esophageal cancer by Fluorouracil Implants in the hospitalized patients in our hospital. Under the direct vision of the endoscopy, the focus is divided into several layers with the interval of 1.0cm in each layer, and one sustained release chemotherapy particle is planted at each interval in 1.0cm. 10mg was implanted at the point of disease, implanted 1 times every 4 weeks and 3 times for 1 courses. All patients were successfully treated in this study. The clinical symptoms remission of the patients before treatment, 3 months after treatment, 3 months after treatment and 6 months after treatment, the change of the lesion area, the improvement of dysphagia, adverse reactions, quality of life, etc. Results: (1) the inhibitory effect of 5- fluorouracil on the cell cycle of human esophageal cancer cell line TE-1 was strongly correlated with the time of drug treatment. The effect on TE-1 was obvious when the concentration was 10 g/ml. The increase of drug concentration could not significantly increase the effect of the drug effect, and the effect of the drug was obviously different when the effect was 96 hours, but the apoptosis was detected. No obvious apoptotic cells were found. (2) the release of Fluorouracil Implants was measured by high performance liquid chromatography (HPLC). It was found that the concentration of the drug released by the slow-release chemotherapy particles increased gradually with the prolongation of the time. After the 48h, the concentration of the drug was basically maintained, and the following concentration decreased slightly. The Fluorouracil Implants treatment of TE-1 cells was found. Different time has different effects. When the number of cells in 48h-72h time section decreases most obviously, the action concentration is about 282 mu g/ml, the longer the action time is more obvious in 96 hours, and it can be clearly detected before the treatment of apoptotic cells (3), compared with the treatment 3,6 months, the difference of the changes of the lesion area and dysphagia classification of the patients is statistically significant. There was no significant difference in the quality of life (P0.05), but there was no significant difference in the quality of life (P0.05, the reason may be that the follow-up time was shorter and the sample was less, not the individual difference), and the difference between the 3 months of white blood cells (WBC) before treatment was statistically significant (P0.05); before treatment, the incidence of complications in the treatment of 3 and 6 months was not unified. Conclusion: 1. Fluorouracil Injection and Fluorouracil Implants can inhibit the proliferation of human esophageal cancer cell lines. The release of.2. Fluorouracil Implants is stable and can promote the apoptosis of human esophageal cancer cell line TE-1, and the longer the time within 96 hours the more obvious the.3. Fluorouracil Implants is fine for esophageal cancer. The inhibitory effect of the cell line is better than that of the Fluorouracil Injection.4. endoscopy with 5- fluorouracil (5-FU) sustained-release particles for the treatment of advanced esophageal cancer. It is a safe, effective, less toxic side effect and a tolerable palliative treatment.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.1

【参考文献】