RNA干扰AKT2的表达对胶质瘤裸鼠移植瘤替莫唑胺敏感性的影响

AKT2-shRNA affects the sensitivity of human glioma cell line U251 xenograft in nude mice to TMZ

目的研究RNA干扰丝/苏氨酸蛋白激酶2(AKT2)对脑胶质瘤移植瘤的替莫唑胺疗效的改变。方法构建胶质瘤裸鼠成瘤模型,瘤内注射和腹腔内给替莫唑胺(TMZ)药物,以替莫唑胺化疗药物和AKT2短发夹结构RNA(AKT2-shRNA)表达载体对成瘤后的裸鼠瘤体进行联合干预治疗,进而观察并测量各组裸鼠肿瘤体积大小。DNA断裂的原位末端标记法(TUNEL)测定并计算分析各组裸鼠成瘤后肿瘤组织细胞的凋亡的变化。结果空白对照组、替莫唑胺化疗组、TMZ+阴性对照组、TMZ+AKT2干扰组裸鼠瘤体体积分别为:(669.34±98.73)mm3、(399.86±55.26)mm3、(383.81±34.01)mm3、(297.72±41.49)mm3;瘤体质量分别为:(1.25±0.26)g、(0.72±0.11)g、(0.69±0.07)g、(0.52±0.07)g。TMZ+AKT2干扰组其肿瘤体积及瘤体质量都明显小于空白对照组、替莫唑胺组和TMZ+阴性对照组,有显著性差异(P<0.05)。TUNEL实验结果显示:空白对照组、替莫唑胺化疗组、TMZ+阴性对照组、TMZ+AKT2干扰组裸鼠瘤体凋亡指数分别为:7.15%±1.04%、25.26%±2.71%、26.63%±3.46%、42.81%±5.97%。TMZ+AKT2干扰组其肿瘤凋亡情况明显高于空白对照组、替莫唑胺组和TMZ+阴性对照组,结果有显著性差异(P<0.05)。结论 RNA干扰AKT2能够显著提高裸鼠胶质母细胞瘤对TMZ化疗的敏感性。

Objective The effects of the expression of AKT2 on sensitivity of glioma cell line U251 to Temozolomide （TMZ） are discussed.MethodsU251 cell implanted tumor model in nude mice was established. By the intratumoral injection and intraperitoneal administration, the nude mice tumor was treated with TMZ chemotherapy drugs and AKT2shRNA expression vector. The nude mice tumor accumulation conditions were observed. DNA in situ end labeling method （TUNEL） was used to analyze the apoptosis of tumor cells in each group.ResultsIn the blank control group, TMZ chemotherapy group, TMZ＋ negative control group, and TMZ＋AKT2 interference group, nude mice tumor volume were （669.34±98.73）mm3, （399.86±55.26）mm3, （383.81±34.01）mm3, （297.72±41.49）mm3, respectively; tumor weight were （1.25±0.26）g, （0.72±0.11）g, （0.69±0.07）g, （0.52±0.07）g, respectively. In TMZ＋AKT2 interference group, the tumor volume and tumor weight were significantly less than those of the blank control group and negative control group, and there was significant difference （P〈0.05）. TUNEL experimental results showed that in blank control group, TMZ chemotherapy group, the TMZ＋negative control group, TMZ＋AKT2 interference body apoptosis index group of tumor were （7.15±1.04）%, （25.26±2.71）%, （26.63±3.46）%, （42.81±5.97）%, respectively. In TMZ＋AKT2 interference group the apoptosis was significantly higher than that of the blank control group and negative control group, and there was significant difference （P〈0.05）.ConclusionRNA interference of AKT2 can significantly improve the sensitivity of glioma chemotherapy to TMZ.