生存素反义寡核苷酸对胃癌细胞株SGC-7901生长的影响

目的　观察生存素蛋白在胃癌细胞株SGC 790 1中的表达及转染不同浓度的生存素反义寡核苷酸(ASODN )对胃癌细胞SGC- 790 1生长的影响。方法　采用细胞免疫组织化学染色观察生存素蛋白在胃癌细胞株SGC- 790 1中的表达。将胃癌细胞株SGC 790 1分为8个组:空白对照组,脂质体转染组,2 0 0nmol/L、40 0nmol/L、60 0nmol/L无义链转染组,2 0 0nmol/L、40 0nmol/L、60 0nmol/L反义链转染组,转染48h后,采用四氮唑盐(MTT )比色法,测定各组的细胞生长抑制率,并采用流式细胞仪检测60 0nmol/L无义链转染组、60 0nmol/L反义链转染组细胞的凋亡率以及生存素蛋白表达。结果　显示生存素蛋白在胃癌细胞株SGC- 790 1的胞核和胞质中均有表达,但以胞核为主;各组生存素ASODN对胃癌细胞株SGC- 790 1有明显的抑制作用(P <0 .0 1) ,抑制作用呈浓度依赖性,60 0nmol/L反义链转染组抑制率为最高,达40 .0 % ,流式细胞仪检测表明生存素ASODN有明显的诱导细胞凋亡的作用,定量检测结果显示生存素ASODN可以下调生存素蛋白至8.8% (P <0 .0 1)。结论　不同浓度的生存素ASODN均能通过下调生存素蛋白表达来抑制胃癌细胞株SGC- 790 1的生长。

The Effect of Antisense Oligonucleotide Targeting Survivin on the Growth of the Gastric Cancer Cell Line SGC-7901

Objective To observe the expression of survivin protein on the gastric cancer cell line SGC-7901 and the effect of different concentration antisense oligonucleotide(ASODN) targeting survivin on the growth of the gastric cancer cell line SGC-7901.Methods Cellular immunohistochemistry was used to observe the expression of survivin protein in the gastric cancer cell line SGC-7901.The cultured cells were divided into 8 groups:control group,lipofectin group,200 nmol/L none sense oligonucleotide(NSODN) group,400 nmol/L NSODN group,600 nmol/L NSODN group,200 nmol/L ASODN group,400 nmol/L ASODN group,600 nmol/L ASODN group.The rate of cell growth inhibition(IR) of all groups were detected by MTT colorimetry after transfected 48 hours.The rate of cells apoptosis with 600 nmol/L NSODN group and 600 nmol/L ASODN group and the expression of survivin protein were detected by flow cytometry on the gastric cancer cell line SGC-7901 quantitively.Results Survivin protein was expressed in karyon and cytoplasm,mainly expressed in karyon.After transfected for 48 hours,the IR of ASODN groups were significantly higher than control group,lipofectin group and oligonucleotide groups(P<0.01).The intensity of inhibition depended with concentration.The inhibiting rate of 600 nmol/L groups reached 40.0%.Flow cytometry demonstrated that ASODN targeting survivin induce apoptosis,survivin protein was 8.8% of control groups(P<0.01).Conclusion ASODN targeting survivin with different concentration can inhibit the growth of the gastric cancer cell line SGC-7901 by decreasing the expression of survivin gene protein.