ADAM17-shRNA 对MCF-7 乳腺癌细胞裸鼠移植瘤的抑制作用研究

In vivo study on MCF-7 cells transfected with ADAM17-shRNA implanted in nude mice

摘要:

目的 观察转染解聚素- 金属蛋白酶17- 短发夹RNA（ADAM17-shRNA）的MCF-7 乳腺癌 细胞在裸鼠体内的生长效果并探索其作用机制。方法 将30 只裸鼠随机分为转染组（接种转染ADAM17- shRNA 的MCF-7 乳腺癌细胞）、空载体组（接种转染shNC 的MCF-7 乳腺癌细胞）及对照组（接种正常 MCF-7 细胞）。分别在各组裸鼠右侧鼷部皮下种植细胞悬液0.2 ml/ 只，观察各组荷瘤裸鼠的生长状态、饮 食及体重变化。第28 天处死裸鼠并取出移植瘤。采用HE 染色观察组织学形态；Western blot 检测肿瘤局 部ADAM17、磷酸化表皮细胞生长因子受体(p-EGFR)、表皮细胞生长因子受体(EGFR)、磷酸化的蛋白激 酶B(p-Akt)、蛋白激酶B(Akt)、磷酸化的细胞外调节蛋白激酶(p-ERK) 及细胞外调节蛋白激酶(ERK) 的 表达。结果 对照组、空载体组及转染组移植瘤最终体积分别为（639.821±15.429）、（643.350±15.543） 和（240.233±10.536）mm3，3 组不同时间点瘤体体积比较有差异（P <0.05）；不同组别瘤体体积比较有差 异（P <0.05）。HE 染色结果显示，移植瘤组织呈乳腺浸润性导管癌特征，对照组与空载体组无明显差异，转 染组较对照组、空载体组坏死多。转染组肿瘤组织中ADAM17、EGFR、p-EGFR、Akt、p-Akt、ERK 及 p-ERK 的蛋白表达水平低于对照组和空载体组（P <0.05）。结论 ADAM17-shRNA 可有效抑制MCF-7 乳 腺癌细胞裸鼠移植瘤的生长。EGFR-PI3K-Akt 和EGFR-MEK-ERK 信号传导通路参与ADAM17-shRNA 的抑制作用。

Abstract:

Objective To explore the effect of ADAM17-shRNA on implanted tumor of MCF-7 breast cancer cells in nude mice. Methods Thirty nude mice were randomly divided into transfection group (MCF-7-ADAM17- shRNA), vector group (MCF-7-shNC) and control group (MCF-7). Three groups of cell suspension were injected subcutaneously in the right side of the footpath of the corresponding nude mice. The general condition of the nude mice was observed every day. When the tumor nodules were measured, the tumor sizes were measured every 4 days. All nude mice were sacrificed by cervical dislocation on the 28th day after implantation. The morphological structures were observed after HE staining, and the expressions of ADAM17, p-EGFR, EGFR, p-AKT, AKT,p-ERK and ERK in three groups were assayed by Western blot. Results The tumor sizes of the control group, the vector group and the transfection group were (639.821 ± 15.429) mm3, (643.350 ± 15.543) mm3 and (240.233 ± 10.536) mm3 respectively. There were differences in tumor volume among different time points in the three groups (P < 0.05). There were differences in tumor volume among the transfection group, the control group and the vector group (P < 0.05). Compared with the transfection group, the necrosis areas in the vector and control groups were lager. The data of Western blot showed that the expressions of ADAM17, p-EGFR, EGFR, p-AKT, AKT, p-ERK and ERK in the transfection group were significantly lower than those in the control and vector groups (P < 0.05). Conclusions ADAM17-shRNA can effectively inhibit the growth of transplanted tumor of human breast cancer MCF-7 cells in nude mice. EGFR-PI3K-AKT and EGFR-MEK-ERK signaling pathways are involved in the process.