Suppression of human lung cancer cell proliferation and metastasis in vitro by the transducer of ErbB-2.1 (T0B1)
Suppression of human lung cancer cell proliferation and metastasis in vitro by the transducer of ErbB-2.1 (T0B1)
复制标题
ErbB-2.1 (T0B1) 转导子对人肺癌细胞增殖和转移的体外抑制作用
DOI：
10.1038/aps.2011.163
复制
发表时间：
2012-02-01
影响因子：
8.2
通讯作者：
Fan, Sai-jun
中科院分区：
文献类型：
--
作者：
Jiao, Yang;Sun, Ke-kang;Fan, Sai-jun
目的：探讨ErbB-2.1（1081）换能器对人肺癌细胞体外增殖、迁移和侵袭的影响。方法：对人肺癌细胞系95-D、A549、NCl-H1299、NCl-H1975、NCl-H661、NCl-H446、NCl-H1395、Calu-3及正常人支气管上皮细胞（HBE）进行检测。Western blot和RT-PCR检测细胞中TOB1的表达水平。采用脂质体诱导的1081重组质粒转染和选择性G418细胞培养，构建了to1过表达细胞株95-D/TOB1。用to1 - sirna转染A549细胞。采用MTT法、流式细胞术和Western blot法检测TOB1对肿瘤细胞增殖和创面愈合的影响。Transwell侵袭性实验评价TOB1对肿瘤细胞迁移和侵袭的影响。明胶酶谱法测定MMP2和MMP9活性。结果：1061在8种人肺癌细胞系中的表达水平明显低于HBE细胞。1081过表达抑制了95-D细胞的增殖，而to1 - sirna敲低to1促进了A549细胞的生长。95-D/TOB1细胞的迁移和侵袭减少，而TOB1的抑制增强了A549细胞的转移。TOB1过表达不仅增加了重要肿瘤抑制因子磷酸酶和紧张素同源物（PTEN）的表达，还调节了PI3K/PTEN信号通路下游效应物，包括Akt、ERK1/2等。相反，1081的表达减少会反过来调节这些因子的表达。TOB1还能调控肺癌细胞中MMP2和MMP9明胶酶的活性。结论：PI3K/PTEN通路可能是TOB1体外调控人肺癌细胞增殖和转移的可能信号通路之一，在肿瘤发生、血管生成和转移过程中起着至关重要的作用。
Aim: To investigate the effects of the transducer of ErbB-2.1 (1081) on the proliferation, migration and invasion of human lung cancer cells in vitro.Methods: Human lung cancer cell lines (95-D, A549, NCl-H1299, NCl-H1975, NCl-H661, NCl-H446, NCl-H1395, and Calu-3) and the normal human bronchial epithelial (HBE) cell line were tested. The expression levels of TOB1 in the cells were determined with Western blot and RT-PCR analyses. TOB1-overexpressing cell line 95-D/TOB1 was constructed using lipofectamine-induced 1081 recombinant plasmid transfection and selective G418 cell culture. The A549 cells were transcend-transfected with TOB1-siRNA. MTT assay, flow cytometry and Western blot analysis were used to examine the effects of TOB1 on cancer cell proliferation and wound healing. Transwell invasive assay was performed to evaluate the effects of TOB1 on cancer cell migration and invasion. The activity of MMP2 and MMP9 was measured using gelatin zymography assay.Results: The expression levels of 1061 in the 8 human lung cancer cell lines were significantly lower than that in HBE cells. 1081 overexpression inhibited the proliferation of 95-D cells, whereas TOB1 knockdown with TOB1-sIRNA promoted the growth of A549 cells. Decreased cell migration and invasion were detected in 95-D/TOB1 cells, and the suppression of TOB1 enhanced the metastasis in A549 cells. TOB1 overexpression not only increased the expression of the phosphatase and tensin homolog (PTEN), an important tumor suppressor, but also regulated the downstream effectors in the PI3K/PTEN signaling pathway, including Akt, ERK1/2, etc. In contrast, decreased expression of 1081 oppositely regulated the expression of these factors. TOB1 also regulates the gelatinase activity of MMP2 and MMP9 in lung cancer cells.Conclusion: The results demonstrate that the PI3K/PTEN pathway, which is essential for carcinogenesis, angiogenesis, and metastasis, may be one of the possible signaling pathways for regulation of proliferation and metastasis of human lung cancer cells by TOB1 in vitro.