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1、复方肠泰抑制大肠癌的作用及机理初探 【毕业论文标题】 复方肠泰抑制大肠癌的作用及机理初探【英文标题】 Primary Study of Fu-Fang-Chang-Tai on the Inhibition to Colon Cancer【中文摘要】 大肠癌(Colorectal Cancer,CRC)是世界范围内最常见的恶性肿瘤之一。本 文通过整体动物和体外细胞生物学实验,从整体动物、组织器官、细胞和分子生物学的不同 水平,观察复方肠泰(FFCT)对人结肠癌细胞 SW480 裸鼠种植瘤的抑制作用;对人结肠癌细胞 SW480 细胞增殖的抑制作用,对其细胞周期、凋亡以及凋亡作用途径进行初步探索;
2、并探索 了对 H22 荷瘤小鼠免疫功能的影响。本文的实验方法如下:通过 FFCT 对人结肠癌细胞 SW480 体外生长的影响以及裸鼠接种 SW480 瘤株后的体内抑瘤作用,进行体内外的抗肿瘤 药效作用研究,并对 FFCT 的临床处方进行筛选;采用噻唑蓝(MTT)比色法测定不同浓度 FFCT 在不同作用时间内对体外培养的 SW480 细胞增殖的影响,同时应用流式细胞术检测其 对细胞增殖周期及凋亡的影响;检测经不同浓度的 FFCT 作用后 SW480 结肠癌细胞内 caspase-3 的活性,比较经 FFCT 作用以及同时经 FFCT 和 caspase-3 抑制剂 Ac-DEVD-CHO 作用后
3、,体外培养的人结肠癌 SW480 细胞凋亡的影响;检测 FFCT 对 H22 荷瘤小鼠瘤重、免 疫器官湿重以及溶血素抗体生成的影响。实验结果表明:“五号处方”的 FFCT 能够显著抑制 人结肠癌细胞 SW480 的增殖;抑制 SW480 裸鼠种植瘤的瘤重,且裸鼠的体重均未见明显减轻,未 见化学药常见的毒性作用。FFCT“处方五”的体内外抑瘤作用优于其它 4 个处方。FFTC 可 在 G0/G1 期以及 G2/M 期阻滞人结肠癌细胞 SW480 的增殖,使 S 期的活细胞比率降低;在一 定范围内,FFCT 的浓度越高、作用时间越长,对肿瘤细胞生长的抑制作用越强,凋亡率也越高。 经 FFCT 培养
4、液分别作用不同时间后的 caspase-3 酶活性,与对照组相比均有显著性变化,呈 现出先增高活性,随后逐渐降低的趋势;加入 caspase-3 抑制剂 Ac-DEVD-CHO 的 FFCT 各浓 度、时间组结肠癌细胞 SW480 的早期凋亡率均有所增加,细胞的坏死率与晚期凋亡率的总 和也有所增加,与空白组比较且呈一定的量-效、时-效关系,但是与没有加入 Ac-DEVD-CHO 时相比,各浓度、时间组的 SW480 早期凋亡率均有所下降,细胞的坏死率与晚期凋亡率的总 和没有显著性变化。FFCT 与抗肿瘤药物环磷酰胺(Cy)有一定的协同作用,抑制瘤重增长;能 明显改善 Cy 化疗后 H22 荷瘤
5、小鼠的脾脏、胸腺等脏器的萎缩,提高胸腺和脾脏的脏器系数, 使因化疗引起的机体免疫功能低下有一定的恢复,并且能够升高 H22 荷瘤小鼠血清溶血素水 平,使 Cy 所致的血清溶血素含量下降有明显恢复,从而可提高 H22 荷瘤小鼠的体液免疫能力。 本文得出如下的结论:FFCT 可抑制大肠癌荷瘤裸鼠的瘤重,并可恢复和增强荷瘤小鼠的免疫 力;FFCT 可抑制体外培养的人结肠癌细胞 SW480 的增殖,其作用机理可能与阻止 G0/G1 期 以及 G2/M 周期的细胞生长、促进 SW480 细胞凋亡相关联。实验结果还提示 caspase-3 途 径可能是 FFCT 促进人结肠癌细胞 SW480 凋亡的重要途
6、径之一,但可能还存在其它作用途 径,有待今后作进一步的深入研究。【英文摘要】 Colorectal cancer is one of the most common cancers in both men and women.To observe the effects of Fu-Fang-Chang-Tai(FFCT) on the growth of SW480 human colon cancer xenografted in athymic mice,and on cell growth,cell generation cycle and apoptosis of human col
7、on carcinoma cell SW480,and on the immune function of mice with carcinoma through in vivo animal experiments and in vitro cell experiments.The mechanisms were explored at levels from whole animal and histological to cytobiological and molecule-biological.The methods were used as follows:The best one
8、 was sieved from several prescriptions of FFCT through the experimental effects on the growth of human colon cancer cells SW480 and SW480 xenografted in athymic mice.The effects of different saturation of FFCT in different action time on cell multiplication of SW480 in vitro culture were determined
9、with methyl thiazolyl tetrazolium(MTT) chromatometry.At the same time the changes of cell generation cycle and apoptosis rate were detected with flow cytometry.Caspase-3 enzyme activities in FFCT-treated SW480 cells were determined. The comparison was made between apoptosis rates in FFCT- treated ce
10、lls and Ac-DEVD-CHO with FFCT-treated cells.The effects of FFCT acting on the tumors, immune organ weight and the formation of hemolysin antibody of the mice with hepatocarcinoma H22 were studied.The results came out as follows:The growth of human colon carcinoma cell SW480 and SW480 xenografted in
11、athymic mice could be significantly inhibited by 5th FFCT, while the body weights of the mice treated by 5th FFCT were not light which compared with the control group.The effectiveness of 5th FFCT was better than other prescriptions. The generation of colon carcinoma cell SW480 in the stage of G0/G1
12、 and G2/M could be interrupted by FFCT.To some extent,the more saturation and time of FFCT acted,the more of inhibitory action on tumor cell growth and apoptosis rate showed.Caspase-3 enzyme activity in FFCT-treated SW480 cells increased at first and then lysised with time went compared with the con
13、trol group.Both apoptosis rates and death rates in Ac-DEVD-CHO with FFCT-treated cells increased significantly compared with the control group.Apoptosis rates in Ac-DEVD-CHO- with FFCT-treated groups were lower than in corresponding FFCT-treated groups while death rates were similar with them. There
14、 were some synergistic actions in Cyclophosphamide united with FFCT to inhibit tumor growth.It could improve the situation of organ atrophy and low formation of hemolysin antibody of the tumor-beating mice caused by chemotherapy in that organism immunity.The humoral immunity could be improved.The co
15、nclusion indicated that:Colon carcinoma could be cured by FFCT through the growth of tumor weight inhibition and the immune function improvement.The growth cycle of tumer cells could be interrupted and cell apoptosis could be induced to inhibit the growth of human colon carcinoma cell SW480 on the e
16、ffect of FFCT.Caspase-3 way was an important but not the only way in the process of SW480 apoptosis.【中文关键词】 复方肠泰; 裸鼠种植瘤; 人结肠癌细胞 SW480; 细胞周期; 细胞凋亡; Caspase-3; 荷瘤小鼠; 免疫 【英文关键词】 Fu-Fang-Chang-Tai; xenografted human colon carcinoma cell; SW480; apoptosis ; caspase-3 ; tumor-bearing mice ; immune function 【论文目录】摘要 5-7 ABSTRACT 7-8 第一章 绪论 14-32 1.1 大肠癌的机理研究 14-19 1.1.1 大肠癌的现代医学发病机理 14-16 1.1.2 中医对大肠癌病因病机的认识 16-19 1.2 中西医药对大肠癌治疗的研究进展 19-23 1.2.1 现代医学研究进展 19-21 1.2.2 中医药
