【作者】 王洪；

【导师】 金先庆；

【作者基本信息】 重庆医科大学 ， 儿科学， 2008， 硕士

【摘要】 第一部分2,6-二异丙基苯酚衍生物对人乳癌细胞的影响目的:观察2,6-二异丙基苯酚衍生物对人乳癌细胞增殖及凋亡的影响方法:体外培养人乳癌细胞株MDA-MB-231,实验分为8组:①正常对照组,②空白对照组,③紫杉醇对照组,2,6-二异丙基苯酚衍生物不同浓度干预组,④50μM,⑤100μM,⑥200μM,⑦400μM,⑧600μM。各实验组干预后继续培养24、48、72小时后待查。采用MTT法选择最佳细胞接种浓度、培养时间及最适抑制浓度,计算其IC50值。观察各组瘤细胞形态变化及增殖情况,采用流式细胞仪观察各组瘤细胞凋亡情况。结果:①MTT法发现人乳癌细胞株MDA-MB-231最佳接种浓度为5.0×10~4/ml,加药时间为接种细胞后24小时。②2,6-二异丙基苯酚衍生物最适抑制浓度为400ug/ml,IC50为390.3 ug/ml。③正常对照组及空白对照组瘤细胞形态及生长增殖情况在各时间段前后无变化。④紫杉醇对照组在48小时后有大量坏死细胞,与2,6-二异丙基苯酚衍生物400μM及600μM浓度组比较,差异不显著(P＞0.05)。⑤各浓度干预组在24小时前后瘤细胞形态及生长增殖情况无变化;200μM浓度组在48小时后可见部分死亡瘤细胞,细胞生长受到抑制,与50μM、100μM组比较,差异显著(P＜0.05);400μM及600μM浓度组在48小时后可见大量死亡瘤细胞,细胞生长受到明显抑制,与50μM、100μM、200μM组间比较,差异非常显著(P＜0.01),400μM与600μM组间差异不显著。结论:2,6-二异丙基苯酚衍生物具有抗人乳癌细胞株MDA-MB-231活性。第二部分2,6-二异丙基苯酚衍生物对人神经母细胞瘤细胞及肝癌细胞的影响目的:观察2,6-二异丙基苯酚衍生物对人神经母细胞瘤及肝癌细胞的影响方法:体外分别培养人神经母细胞瘤细胞株和肝癌细胞株。把第一部分紫杉醇对照组换为羟基喜树碱对照组,其余实验分组方法、对照组及干预组浓度设计、时段设置和观察指标同第一部分。结果:①MTT法发现人神经母细胞瘤细胞株及肝癌细胞株最佳接种浓度为5.0×10~4/ml,加药时间为接种细胞后24小时。②神经母细胞瘤最适抑制浓度为200ug/ml,IC50为268.2 ug/ml。肝癌细胞最适抑制浓度为200ug/ml,IC50为185.5ug/ml。③正常对照组及空白对照组瘤细胞形态及生长增殖情况在各时间段两种肿瘤细胞均无变化。④神经母细胞瘤细胞组48小时后,200μM、400μM、600μM组吸光度值与50μM、100μM组比较有非常显著性差异(P＜0.01)。200μM与400μM组比较无显著性差异(P＞0.05),400μM与600μM组比较有显著性差异(P＜0.05)。羟基喜树碱对照组与200μM与400μM组比较无显著性差异(P＞0.05),与600μM组比较有非常显著性差异(P＜0.01)。⑤肝癌细胞组48小时后,50μM组与正常对照组和空白对照组吸光度值比较无显著性差异(P＞0.05)。100μM、200μM、400μM、600μM组吸光度值与50μM组、对照组比较有非常显著性差异(P＜0.01)。100μM、200μM、400μM组与600μM组比较有非常显著性差异(P＜0.01)。羟基喜树碱对照组与100μM与200μM组比较无显著性差异(P＞0.05),与400μM组比较有显著性差异(P＜0.05),与600μM组比较有非常显著性差异(P＜0.01)。⑥两种肿瘤细胞在100μM浓度组间比较,差异非常显著(P＜0.01)。结论:2,6-二异丙基苯酚衍生物具有抗人神经母细胞瘤和肝癌细胞活性,但对抗肝癌细胞表现更佳。更多还原

【Abstract】 PART 1 EFFECTS OF 2,6-DIISOPROPYL PHENOL DERIVATIVES ON HUMAN BREAST CANCER CELLSObjective:To observe the effects of 2,6-diisopropyl phenol derivatives on proliferation and apoptosis of human breast cancer cells.Methods:Breast cancer cell lines MDA-MB-231 were cultured in vitro.There were 8 groups in the experiment:cell control group,solvent control group(0.5%DMSO),paclitaxel control group,different concentration experimental group of 2,6-diisopropyl phenol derivatives at 50μM,100μM,200μM,400μM and 600μM.Each experimental group continued to be cultured for 24 hours after intervention in order to be investigated.Optimal cell innovulation concentration and culture time were chosen with the application of MTT assay;detecting morphologic change and proliferation condition of tumor cells in each group and then observing every group’s apoptosis of tumor cells with flow cytometer.Results:(1)The optimal innoculation concentration of human breast cancer cell line MDA-MB-231 was found to be 5.0×10~4/ml and the administration timing of drug should be 24 hours after inoculating cells with MTT assay. (2)There was no morphologic change or proliferation condition change of tumor cells in cell control and solvent control groups within 24 hours.(3)Lots of necrotic cells were observed in hydrocamptothecin control group,compared with 2,6-diisopropyl phenol derivatives at 400μM and 600μM concentration groups,the difference was not significant (P＞0.05).(4)No morphologic change or proliferation condition change of tumor cells was found in 2,6-diisopropylphenol derivatives at 50μM and 100μM concentration groups within 24 hours.Part of dead tumor cells were observed and some cells underwent apoptosis in 200μM concentration group,compared with 50μM and 100μM groups,the difference was remarkable(P＜0.05);In 400μM and 600μM concentration groups,there were a big amount of dead tumor cells after 24 hours,lots of tumor cells underwent apoptosis,compared with 50μM,100μM and 200μM groups,the differences were quite significant(P＜0.01),group comparsion between 400μM and 600μM showed that the difference was not significant.Conclusion:2,6-diisopropylphenol derivatives have the activity to resist human breast cancer cell line MDA-MB-231. PART 2 EFFECTS OF 2,6-DIISOPROPYLPHENOL DERIVATIVES ON HUMAN NEUROBLASTOMA CELLS AND HEPATOMA CARCINOMA CELLSObjective:To detect effects of 2,6—diisopropylphenol derivatives on human neuroblastoma cells and hepatoma carcinoma cells.Methods:Human neuroblastoma cell lines and hepatoma carcinoma cell lines were cultured in vitro sepreately.Paclitaxel control group in part 1 changed to hydroxycamptothecin control group in part 2.The other of experimental groups,concentration and time controlled,and detecting part same as part 1.Results:(1)The optimal innoculation concentration of human neuroblastoma cell lines and hepatoma carcinoma cell lines was found to be 5.0×10~4/ml and the administration timing of drug should be 24 hours after inoculating cells with MTT assay.(2)There was no morphologic change or proliferation condition change of tumor cells in cell control and solvent control groups within 24 hours.(3)Absorbance values of neuroblastoma cell group,200μM,400μM and 600μM groups had significant difference with that of 50μM and 100μM groups(P＜0.01).Group comparsion between 200μM and 400μM groups showed no sifnificant difference(P＞0.05).Comparsion between 400μM and 600μM groups had obvious difference.Compared with 200μM and 400μM groups,hydroxycamptothecin control group had no remarkable difference(P＞0.05),however,when compared with 600μM group,the difference was significant(P＜0.01)(4)Hepatoma carcinoma cell group and 50μM group had no significant difference with cell control group and solvent control group in absorbance values(P＞0.05).Absorbance values of 100μM,200μM,400μM and 600μM groups had very significant difference with that of 50μM group and contol group(P＜0.01).There was significant difference between 600μM group and 100μM,200μM,400μM groups(P＜0.01).Compared with 100μM and 200μM groups,hydroxycamptothecin control group had no obvious difference(P＞0.05),when compared with 400μM group,the difference was significant(P＜0.05),as compared with 600μM group,the difference was quite remarkable(P＜0.01)(5)Group comparsion between two kinds of tumor cells at 100μM concentration showed significant difference(P＜0.01)Conclusion:2,6-diisopropyl phenol derivatives have the activity to resist human neuroblastoma cells and hepatoma carcinoma cells,but the effect of anti-hepatoma carcinoma cell is better.更多还原