膜联蛋白Ⅱ反义寡核苷酸对人膀胱癌BIU-87细胞膜联蛋白Ⅱ表达及细胞侵袭力的影响

Effect of AnnexinⅡ ASODN on the expression of AnnexinⅡgene and invasion of human bladder cancer BIU-87 cell

目的探讨膜联蛋白Ⅱ(AnnexinⅡ)反义寡核苷酸(ASODN)对膀胱癌BIU-87细胞AnnexinⅡ基因表达及侵袭力的影响。方法设计合成AnnexinⅡASODN,采用脂质体将低、中及高浓度(6、12及18μmol/L)的AnnexinⅡASODNs分别转染BIU-87细胞24、48及72 h,同时设空白对照组(不转染)。采用免疫细胞化学与原位杂交法分别检测各组细胞中AnnexinⅡ蛋白与m RNA的表达;采用MTT法检测细胞增殖情况,计算细胞生长抑制率,采用Boyden chamber侵袭实验检测高浓度转染72 h细胞侵袭力情况。结果与空白对照组相比,各转染组细胞AnnexinⅡ蛋白和m RNA的表达均减弱,且差异均有统计学意义(P<0.05),且AnnexinⅡ蛋白及m RNA的表达随AnnexinⅡASODN浓度的增加及转染时间的延而减弱(P<0.05);AnnexinⅡASODN转染组细胞生长抑制率明显增加(P<0.05);细胞穿越Matrigel的细胞数交空白组减少,且差异均有统计学意义(P<0.05)。结论 AnnexinⅡASODN可有效抑制膀胱癌BIU-87细胞AnnexinⅡ蛋白及m RNA的表达,抑制BIU-87细胞的生长及侵袭力。

[Objective]To explore the effect of AnnexinⅡ ASODN on the expression of AnnexinⅡ gene and invasion in human bladder cancer BIU-87 cells.[Methods]The oligonucleotide of AnnexinⅡASODN were designed firstly. Three different concentrations of Annexin Ⅱ ASODN（6, 12 and 18 μmol/L was used to transfect BIU-87 cells with different time exposure（24, 48 and 72 h）, and the control group were established accordingly by no transfection. The level of m RNA and protein of Annexin Ⅱ in the 6 groups were measured by insitu hybridization and immunocytochemistry. MTT method was used to detect the cell prolife ration and Boyden chamber invasion was used to detect t the BIU-87 cell invasion. [Results]Compared with the control groups, there were significant differences in the expressions of AnnexinⅡ m RNA and protein of BIU-87 cells among the 3 groups at 24, 48, and 72 h（P〈0.05）, and increased with the increase of si RNA concentration and time exposure（P〈0.05）. Compared with the control groups, the growth inhibition rate of BIU-87 cells transfected with AnnexinⅡASODN for different time exposure was higher（P〈0.05）, and increased with the increase of ASODN concentration and time exposure（P〈0.05）. Compared with the control group, AnnexinⅡ ASODN group can inhibit the invasion force of bladder cancer BIU-87 cell,and the differences were statistically significant（P〈0.05）.[Conclusion]AnnexinⅡASODN could effectively knock down the expression of Annexin Ⅱ, which could significantly inhibit the invasion force of bladder cancer BIU-87 cell.