抑制膜联蛋白A2表达对人胃癌细胞株裸鼠移植瘤生长的影响及机制

Effect of ANXA2 on Growth of Implanted Human Gastric Cancer in Nude Mice

[目的]通过体内研究探讨膜联蛋白A2(ANXA2)对胃癌生长的影响及相关作用机制。[方法]常规培养人胃癌细胞株BGC-823;将细胞接种到裸鼠皮下制备移植瘤模型。制模成功后随机分组,分别以脂质体/ANXA2-siRNA复合物(实验组)、脂质体/Non-siRNA复合物(对照组)或生理盐水(空白组)分别在各组肿瘤局部注射,每4d干预1次。干预4次后处死,绘制肿瘤生长曲线并比较瘤重。应用荧光定量RT-PCR及免疫组化技术检测各组ANXA2、增殖细胞核抗原(PCNA)、p27m RNA和蛋白表达。[结果]裸鼠皮下移植瘤造模成功率100.00%(15/15)。实验组肿瘤生长比其他两组缓慢,最终体积实验组为(1680.26±110.51)mm3,对照组为(2194.18±188.48)mm3,空白组(2172.37±212.36)mm3(F=9.341,P=0.004);肿瘤重量实验组(20.63±0.46)g,对照组为(22.92±0.86)g,空白组为(23.44±0.90)g(F=11.688,P=0.002)。PCR及免疫组化结果显示实验组ANXA2、PCNAm RNA和蛋白表达明显低于其他两组(P<0.05),而p27m RNA和蛋白表达明显高于其他两组(P<0.01)。[结论 ]抑制ANXA2基因表达后胃癌移植瘤的生长明显受到抑制,这可能与ANXA2基因调控PCNA、p27表达有关。

[Objective] To investigate the effect of Annexin A2（ANXA2） on the growth of gastric cancer in vivo. [Methods] Human gastric cancer BGC-823 cells were inoculated into 15 nude mice subcutaneously to establish gastric cancer model. The cancer-bearing nude mice were divided into 3 groups randomly with 5 in each group,the animals were treated with liposome/ANXA2-siRNA compound（ANXA2 group）,liposome/non-siRNA compound（control group） or normal saline（blank group）,respectively. After nude mice were sacrificed,volume and weight of tumors were measured,and the expressions of ANXA2,PCNA,p27 m RNA and protein were detected with fluorescence quantitative RT-PCR and immunohistochemistry,respectively. [Results] The tumor growth in ANXA2 group was slower than that in other 2 groups; and the final volume of tumors was（1680.26±110.51）mm3in ANXA2 group,（2194.18±188.48）mm3in control group and（2172.37±212.36）mm3in blank group（F =9.341,P =0.004）. The weight of tumors was（20.63 ±0.46）g in ANXA2 group,（22.92±0.86）g in control group and（23.44 ±0.90）g in blank group（F=11.688,P=0.002）. The expressions of ANXA2 and PCNA m RNA and protein in ANXA2 group were lower than those in other 2 groups,whereas expressions of p27 m RNA and protein in ANXA2 group were higher than those in the other 2 groups（P〈0.01）. [Conclusion] The growth of tumor can be inhibited after ANXA2 inhibition,indicating that ANXA2 may regulate expressions of PCNA and p27 in gastric cancer.