【作者】 陶慧；

【导师】 刘淑萍； 王文瑞；

【作者基本信息】 内蒙古医学院 ， 免疫学， 2008， 硕士

【摘要】 目的通过研究在不同浓度蒙药巴特日作用下,胃癌MGC-803细胞内端粒酶表达变化,探讨蒙药巴特日抑制肿瘤生长是否与端粒酶活性有关。为其临床应用提供一定的实验依据。方法将蒙药巴特日以细胞培养液稀释至终浓度,分别处理体外培养的人胃癌MGC-803细胞株,采用细胞毒敏感性实验MTT法检测蒙药巴特日对人胃癌MGC-803细胞的生长抑制作用;采用端粒重复扩增-银染法（TRAP-银染法）测定端粒酶活性;采用逆转录-聚合酶链式反应（RT-PCR）法观察端粒酶逆转录酶hTERT基因的表达。结果（1）MTT结果显示,蒙药巴特日可明显抑制胃癌MGC-803细胞的生长,与对照组相比其生长抑制率有显著性差异（P＜0.05）其IC₅₀为1mg/ml。（2）胃癌MGC-803细胞分别给予蒙药巴特日浓度为0.3 mg/ml（a组）,0.5mg/ml（b组）,1mg/ml（c组）,1.25mg/ml（d组）,2.5mg/ml（e组）,空白对照（f组）作用48小时后,随着药物浓度的增加,端粒酶活性阳性表达呈下降趋势。（3）RT-PCR结果显示,不同浓度蒙药巴特日处理细胞48h后,hTERTmRNA表达与对照组相比明显降低（P＜0.05）。结论1.蒙药巴特日可明显抑制胃癌MGC-803细胞的生长,其IC₅₀为1mg/ml。2.蒙药巴特日能下调胃癌MGC-803细胞端粒酶活性,且有剂量依赖性。3.蒙药巴特日能下调胃癌MGC-803细胞hTERTmRNA表达,且有剂量依赖性。更多还原

【Abstract】 Objective The purpose for our test is to study the effect of telomerase activity in MGC -803 human gastric cancer cell line by BaTeRi after incubation in different concentrations and to explore its anti - neoplasm mechanism.The important basis for applying BaTeRi in testing gastric cancer was also provided.Method MTT assay was used to determine the inhibition of MGC - 803 cell growth after incubation in different concentrations of BaTeRi. Telomerase activity was examined by Telomerase Regulation- associated Protein- polymerase Chain Reaction（TRAP- PCR）-silver staining.Expression of hTERTmRNA in MGC- 803 cells were detected by RT- PCR.Result（1）BaTeRi inhibited significantly the growth of MGC-803 human gastric cancer cell line and elevated the inhibiting rate（P＜0. 05）as compared with control groups.The IC₅₀was 1mg/ml.（2）MGC - 803 human gastric cancer cell line was divided into 5 groups and treated with BaTeRi at the concentration of 0. 3mg/ml（group a）,0.5mg/ml（group b）,1mg/ml（group c）,1.25mg/ml（group d）,2.5mg/ ml（group e）,control group（group f）,respectively.After treated 48 hours,the telomerase activation showed the tendency of declining with the increase of BaTeRi concentration.（3） Compared to control group,BaTeRi could down-regulate the expression of hTERTmRNA in MGC- 803 cells significantly（P＜0.05）after treated with different concentrations of BaTeRi for 48 hours.Conclusion（1）BaTeRi inhibited significantly the growth of MGC-803 human gastric cancer cell line.The IC₅₀was 1mg/ml.（2）BaTeRi could down- regulate telomerase activity of MGC- 803 human gastric cancer cell line and was dependant on the concentration of BaTeRi.（3）BaTeRi could down- regulate expression of hTERTmRNA of MGC - 803 human gastric cancer cell line and was dependant on the concentration of BaTeRi更多还原