丹参酮ⅡA联合5-FU对低氧下人胃癌SGC7901细胞增殖、凋亡的影响及与HIF-1α和突变型P53表达的关系  

Effects of combined Tan ⅡA and 5-FU on cell proliferation,apoptosis,and the expression of HIF-1α and mutant P53 in human gastric cancer cell line SGC7901 under hypoxia

作　　者：邢国辉[1] 冯玉光[2] 宗绪山[1] 吴美英[1] 朱芸[1] 

机构地区：[1]潍坊医学院,山东省潍坊市261042 [2]潍坊医学院附属医院消化内科,山东省潍坊市261042

出　　处：《世界华人消化杂志》2010年第3期222-228,共7页World Chinese Journal of Digestology

基　　金：山东省中医管理局中医药科技发展计划基金资助项目;No.2007-156~~

摘　　要：目的:观察低氧环境下不同浓度的丹参酮ⅡA(TanⅡA)联合5-氟尿嘧啶(5-FU)对人胃癌SGC7901细胞增殖、凋亡的影响及与HIF-1α和突变型P53的表达.方法:用氯化钴(CoCl2)创建低氧模型,采用0、0.5、1.0、2.0、5.0、10.0mg/L的TanⅡA联合10.0mg/L的5-FU分别作用于低氧SGC7901细胞24、48和72h,MTT法检测细胞活力;用上述同样方式作用于低氧SGC7901细胞24、48和72h后,Hoechst染色法检测细胞凋亡;TanⅡA(0、0.5、2.0、10.0mg/L)联合10.0mg/L的5-FU作用于低氧SGC7901细胞48h后,免疫细胞化学二步法检测HIF-1α及突变型P53的表达.结果:低氧环境下,不同浓度的TanⅡA联合10.0mg/L5-FU呈时间、剂量依赖性地抑制SGC7901细胞的增殖(均P<0.01),10.0mg/LTanⅡA联合5-FU作用细胞72h后,其抑制率为67.46%.0.5-10.0mg/LTanⅡA联合5-FU作用细胞24、48、72h,TanⅡA呈时间、剂量依赖性地促进SGC7901细胞凋亡(均P<0.01).HIF-1α及突变型P53表达明显高于常氧组(t=22.786,13.914,均P<0.01),不同浓度的TanⅡA联合10.0mg/L5-FU作用细胞48h后,HIF-1α、突变型P53蛋白表达明显降低(F=182.234,130.062,均P<0.01),且二者呈高度正相关(n=5,r=0.995,P<0.01).结论:在低氧环境下,TanⅡA可能通过抑制HIF-1α及突变型P53蛋白表达从而增强5-FU抑制SGC7901细胞增殖及诱导凋亡作用.AIM：To investigate the effects of combined tanshinone ⅡA（Tan ⅡA） and 5-fluorouracil（5-FU） on cell proliferation,apoptosis,and the expressions of hypoxia-inducible factor-1 alpha（HIF-1α） and mutant P53（mt P53） in human gastric cancer cell line SGC7901 under hypoxia.METHODS：Hypoxia was induced in SGC7901 cells by cobalt dichloride treatment.SGC7901 cells under hypoxia were treated with different concentrations of Tan ⅡA in combination with 10.0 mg/L of 5-FU for 24,48 and 72 hours.Cell proliferation was detected by methyl thiazolyl tetrazolium（MTT） assay.Cell apoptosis was detected by Hoechst staining.The expression of HIF-1α and mt P53 proteins was detected by immunocytochemistry.RESULTS：Combined Tan ⅡA and 5-FU signif icantly inhibited the proliferation of SGC7901 cells（all P 0.01） in a dose-and time-dependent manner under hypoxia.The reduced proliferation rate of cells incubated with Tan ⅡA at a concentration of 10 mg/L and 5-FU for 72 hours was 67.46%.Hoechst staining showed that Tan ⅡA in combination with 5-FU promoted the apoptosis of SGC7901 cells in a dose-and time-dependent manner under hypoxia（all P 0.01）.Immunocytochemical staining revealed that the expression levels of HIF-1α and mt P53 proteins in SGC7901 cells under hypoxia were evidently higher than those in SGC7901 cells under normal conditions（t=22.786 and 13.914,respectively;both P 0.01）.However,Tan ⅡA in combination with 5-FU signif icantly downregulated the expression of HIF1α and mt P53 proteins in SGC7901 cells under hypoxia（F=182.234 and 130.062,respectively;both P 0.01）.A signif icant positive correlation was noted between the expression of HIF-1α and mt P53 in SGC7901 cells（n=5,r=0.995,P 0.01）.CONCLUSION：Tan ⅡA can significantly enhance 5-FU-mediated growth inhibition and apoptosis induction in SGC7901 cells under hypoxia perhaps by downregulating HIF-1α and mt P53 protein expression.

关 键 词：丹参酮ⅡA 5-氟尿嘧啶 低氧 胃癌细胞 凋亡 P53 低氧诱导因子-1Α 

分 类 号：R735.2[医药卫生—肿瘤] Q75[医药卫生—临床医学]