Inhibitory effects of adenovirus mediated Akt1 and PIK3R1 shRNA on the growth of malignant tumor cells in vitro and in vivo
Inhibitory effects of adenovirus mediated Akt1 and PIK3R1 shRNA on the growth of malignant tumor cells in vitro and in vivo
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DOI：
10.4161/cbt.8.11.8285
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发表时间：
2009-06-01
影响因子：
3.6
通讯作者：
Wang, Tao
中科院分区：
文献类型：
--
作者：
Fu, Yanchao;Zhang, Qingyu;Wang, Tao
磷脂酰肌醇3-激酶(PI3K)/Akt在许多恶性肿瘤的形成中发挥着关键作用，并已被证明是重要的治疗靶点。在本研究中，使用靶向人Akt1和PIK3R1序列的小发夹RNA（shRNA）表达构建体来检测对SGC7901胃腺癌细胞和U251神经胶质瘤细胞的增殖和侵袭抑制作用。 MTT分析显示，shRNA处理组细胞生长被抑制超过60%，并伴有G(1)/G(0)期停滞，表明细胞生长活性较差。与对照组（105+/-4.0）和无义序列组（102.5+/-6.4）相比，shRNA处理组侵入基质胶的细胞数量显着减少（51.6+/-3.9）。此外，shRNA处理的SGC7901皮下裸鼠模型中肿瘤体积明显小于对照组和无义序列组。当 Akt1 和 PIK3R1 显着下调时，增殖细胞核抗原 (PCNA)、CyclinD1 和基质金属蛋白酶 (MMP-2、MMP-9) 下调，而金属蛋白酶组织抑制剂 -2 (TIMP-2) 和 p53 上调。我们的结果表明，针对 Akt1 和 PIK3R1 的 shRNA 以序列特异性方式显着下调其表达，对 SGC7901 和 U251 细胞发挥增殖和侵袭抑制作用。总之，我们的数据提出了一种调节恶性肿瘤细胞生长的新机制，并为恶性肿瘤的新组合基因疗法提供了证据。
Phosphatidylinositol 3-kinase (PI3K)/Akt plays a critical role in the formation of many malignant tumors, and has been shown to be an important therapeutic target. In the present study, small hairpin RNA (shRNA) expression constructs that target sequences of human Akt1 and PIK3R1 were used to examine the proliferation and invasion inhibition effects on SGC7901 gastric adenocarcinoma cells and U251 glioma cells. Cell growth was inhibited by over 60%, as indicated by a MTT assay, and was accompanied by G(1)/G(0) phase arrest in the shRNA treated group, indicating poor cell growth activities. The number of cells invading through the matrigel in the shRNA treated group were significantly decreased (51.6 +/- 3.9) compared with that of the control group (105 +/- 4.0) and the nonsense sequence group (102.5 +/- 6.4). In addition, the tumor volumes in the SGC7901 subcutaneous nude mouse model treated with shRNA were significantly smaller than those of the control group and nonsense sequence group. When Akt1 and PIK3R1 were dramatically downregulated, proliferating cell nuclear antigen (PCNA), CyclinD1 and matrix metalloproteinases (MMP-2, MMP-9) were downregulated, while tissue-Inhibitor of Metalloproteinase-2 (TIMP-2) and p53 were upregulated. Our results demonstrated that shRNA targeting Akt1 and PIK3R1 downregulates their expression significantly in a sequence-specific manner, exerting proliferation and invasion inhibition effects on SGC7901 and U251 cells. In conclusion, our data suggests a novel mechanism for the regulation of malignant tumor cell growth and provides evidence for new combinatory gene therapy for malignant tumors.