兰州大学机构库 >生命科学学院
6-(对氯苯基-3-1-对氯苯基-5-甲基-1,2,3-三唑-4-基)-s-三唑[3,4-b]-1,3,4-噻二唑抗肿瘤活性及机理的研究
Alternative TitleStudy on antitumor activities and mechanisms of 6-(p-chlorophenyl)-3-[1-(p-chlorophenyl)-5-methyl-1H-1,2,3-triazol-4-yl]-1,2,4-triazolo[3,4-b]-1,3,4-thiadiazole
张琪
Thesis Advisor王勤
2004-05-10
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keywords-三唑[3,4-b]-1,3,4-噻二唑 抗肿瘤活性 再分化
Abstract目的:从体内外研究6-(对氯苯基-3-1-对氯苯基-5-甲基-1,2,3-三唑-4-基)-s-三唑[3,4-b]-1,3,4-噻二唑(Compd. I)的抗肿瘤活性,深入探讨其作用机理。方法:通过MTT法进行Compd. I体外抗癌活性初探,测定IC50值;台盼蓝拒染法绘制生长曲线;扫描电镜、细胞电泳迁移率、甲胎蛋白(AFP)和H3-TdR掺入法研究对人肝癌细胞SMMC-7721增殖的再分化机理;荧光双染、流式细胞术测定对细胞凋亡形态及细胞周期;单细胞电泳法测定对细胞DNA的损伤。建立小鼠体内S180移植性肿瘤模型进行体内抗癌活性的研究。腹腔注射不同剂量的Compd. I (100 mgּkg-1、45 mgּkg-1、10 mgּkg-1),连续注射8d后测定其抑瘤率,荷瘤小鼠免疫器官和免疫功能的影响:荷瘤小鼠脾淋巴细胞增殖的变化;自然杀伤细胞(NK)杀伤功能和血清蛋白含量的变化。在组织学水平观察对实体瘤的作用。结果:MTT法测得Compd. I在72 h SMMC-7721的IC50 为35.22 μgּmL-1;扫描电镜,细胞电泳和H3-TdR掺入法,发现Compd. I在低浓度(8 μgּmL-1、16 μgּmL-1)时,可明显抑制肿瘤细胞的增殖,使细胞表面微绒毛脱落,细胞表达AFP水平降低,电泳迁移率降低,DNA复制减慢,说明癌细胞恶性程度降低,促使SMMC-7721细胞再分化;较高浓度(20 μgּmL-1、40 μgּmL-1)的Compd. I处理后,在荧光显微镜下可观察到染色体凝聚,凋亡小体和凋亡细胞的出现;流式细胞术结果显示浓度为80 μgּmL-1时,出现凋亡峰,凋亡率达到4.2%,单细胞凝胶电泳图谱显示细胞随着浓度的增加尾迹增长。这些结果均提示在较高浓度Compd. I处理后细胞出现了凋亡,并且对SMMC-7721细胞的DNA产生了损伤。以上结果表明:Compd. I抑癌的机制是低浓度诱导SMMC-7721细胞再分化进而在高浓度下促使癌细胞凋亡。Compd. I在体内对S180肿瘤的生长有明显的抑制作用,且具有剂量效应关系,但敏感性不同,剂量为100 mgּkg-1时,抑瘤率可达到69.08 %,病理切片显示Compd. I可在一定程度上促进肿瘤坏死;对荷瘤小鼠免疫功能的测定显示:Compd. I对荷瘤小鼠的脾脏指数和胸腺指数影响较小,高浓度(100 mg.kg-1)时还可提高脾指数,45mgּkg-1时可提高NK细胞杀伤功能,至正常的95.4%。结论:Compd. I具有明显的抗肿瘤活性,体外可以诱导人肝癌细胞SMMC-7721的再分化,高浓度下可产生部分凋亡;体内抑瘤作用明显,能提高机体免疫细胞对肿瘤的杀伤活性。
Other AbstractThe antitumor activity and its mechanism of one new synthesized compound (Compd. I), derivative of thiadiazol, was studied. In vitro, the inhibition of Compd. I on SMMC-7721 cells were tested by MTT colorimetric assay and trypan blue exclusion test, and the effect of Compd. I on redifferentiation of cancer cells was analyzed by scanning electronic microscope, cell electrophoresis, AFP and isotope labeling technique (H3-TdR). Our results showed that Compd. I could inhibit the proliferation of human hepatoma cells SMMC-7721 in the dose and time-dependent manners. Treatment with Compd. I at the concentration of 8 μgּmL-1and 16 μgּmL-1, the average of AFP decreased from 205.14±6.41 ng.mg-1Pr to 115.68±3.47 and 78.57±2.35 ng.mg-1Pr; electrophoresis rate of cells reduced from 2.14 μmּs-1ּV-1ּcm-1 of control to 1.54 μmּs-1ּV-1ּcm-1, DNA replication decreased and the microvilli on the surface of SMMC-7721 cells reduced obviously. These results indicate that Compd. I may inhibit proliferation of cancer cells by reversing SMMC-7721 cells malignant phenotypic characteristics and inducing redifferentiation. Observing with fluorescence microscope, we found that treatment with Compd. I at the concentration of 20 μgּmL-1 and 40 μgּmL-1, DNA agglomerated and apoptotic bodies appeared. The apoptotic rate of SMMC-7721 cells was assayed by flow cytometry and the result indicated the apoptotic rate was 4.2 % when SMMC-7721 cells were treated with 80 μgּmL-1 Compd. I. Comet electrophoresis results showed a comet feature. These results indicated that high concentration of Compd. I (20 μgּmL-1, 40 μgּmL-1 and 80 μgּmL-1) can induce DNA damage and apoptosis in SMMC-7721 cells. In vivo, the inhibition rate of Compd. I on S180 tumor in the animal models was measured, and the influences of Compd. I on immune organs and immune functions in tumor-bearing mice were determined. Detection of S180 tumor growth inhibition showed that Compd. I reduced the tumor size, and the inhibition reached 69.08 %. Pathological slices showed the necrosis, invasion and inflammation in S180 tumor, and after treatment with Compd. I, necrosis of tumor organ increased. Moreover, Compd. I could protect and activate immune system of tumor-bearing mice, such as increase the splenetic index, activate NK cell and T cell. All these results indicated that Compd. I could inhibit the proliferation of tumor in vitro a...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222086
Collection生命科学学院
Recommended Citation
GB/T 7714
张琪. 6-(对氯苯基-3-1-对氯苯基-5-甲基-1,2,3-三唑-4-基)-s-三唑[3,4-b]-1,3,4-噻二唑抗肿瘤活性及机理的研究[D]. 兰州. 兰州大学,2004.
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