RNA interference reveals that ligand-independent met activity is required for tumor cell signaling and survival
RNA interference reveals that ligand-independent met activity is required for tumor cell signaling and survival
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DOI：
10.1158/0008-5472.can-04-1043
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发表时间：
2004-11-01
期刊：
影响因子：
11.2
通讯作者：
Woude, GFV
中科院分区：
文献类型：
--
作者：
Shinomiya, N;Gao, CF;Woude, GFV
肝细胞生长因子/分散因子-Met 信号传导与肿瘤生长、侵袭和转移有关。通过靶向 Met 蛋白酪氨酸激酶来抑制该信号通路可能是抑制恶性肿瘤生长的理想策略。利用 RNA 干扰技术和携带针对小鼠、犬和人类 Met 的小干扰 RNA 构建体（Ad Met 小干扰 RNA）的腺病毒载体，我们可以敲低 c-met mRNA。我们在配体依赖性和配体非依赖性小鼠、犬和人类肿瘤细胞系中均表现出对 Met 的显着依赖性。小鼠乳腺肿瘤 (DA3) 细胞和 Met 转化的 NIH3T3 (M114) 细胞，以及人类和犬前列腺癌（PC-3 和 TR6LM、人类肉瘤 (SK-LMS-1)、胶质母细胞瘤 (DBTRG) 和胃癌 (MKN45) 细胞，在感染 Ad Met 小干扰 RNA 后，Met 表达均显着降低。在这些细胞中，我们观察到肿瘤细胞受到抑制重要的是，Ad Met 小干扰 RNA 会导致许多肿瘤细胞系（尤其是 DA3 和 MKN45）细胞凋亡，但不会对 MDCK 犬肾细胞产生不利影响。 p44/42 丝裂原激活蛋白激酶。我们进一步表明，e-met 小干扰 RNA 腺病毒的瘤内感染可导致肿瘤生长大幅减少，因此，Met 小干扰 RNA 腺病毒是研究 Met 功能的可靠工具，并提高了其应用于癌症治疗的可能性。
Hepatocyte growth factor/scatter factor-Met signaling has been implicated in tumor growth, invasion, and metastasis. Suppression of this signaling pathway by targeting the Met protein tyrosine kinase may be an ideal strategy for suppressing malignant tumor growth. Using RNA interference technology and adenovirus vectors carrying small-interfering RNA constructs (Ad Met small-interfering RNA) directed against mouse, canine, and human Met, we can knock down c-met mRNA. We show a dramatic dependence on Met in both ligand-dependent and ligand-independent mouse, canine, and human tumor cell lines. Mouse mammary tumor (DA3) cells and Met-transformed NIH3T3 (M114) cells, as well as both human and canine prostate cancer (PC-3 and TR6LM, human sarcoma (SK-LMS-1), glioblastoma (DBTRG), and gastric cancer (MKN45) cells, all display a dramatic reduction of Met expression after infection with Ad Met small-interfering RNA. In these cells, we observe suppression of tumor cell growth and viability in vitro as well as inhibition of hepatocyte growth factor/scatter factor-mediated scattering and invasion in vitro, whether Met activation was ligand dependent or not. Importantly, Ad Met small-interfering RNA led to apoptotic cell death in many of the tumor cell lines, especially DA3 and MKN45, but did not adversely affect MDCK canine kidney cells. Met small-interfering RNA also abrogated downstream Met signaling to molecules such as Akt and p44/42 mitogen-activated protein kinase. We further show that intratumoral infection with e-met small-interfering RNA adenovirus results in a substantial reduction in tumor growth. Thus, Met small-interfering RNA adenoviruses are reliable tools for studying Met function and raise the possibility of their application for cancer therapy.