School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, India.
Cell Signal. 2011 Oct;23(10):1563-77. doi: 10.1016/j.cellsig.2011.04.009. Epub 2011 May 6.
Cell migration is the hallmark of cancer regulating anchorage independent growth and invasiveness of tumor cells. Hyaluronan (HA), an ECM polysaccharide is shown to regulate this process. In the present report, we demonstrated, supplementation of purified recombinant hyaluronan binding protein 1(HABP1/p32/gC1qR) from human fibroblast cDNA enhanced migration potential of highly invasive melanoma (B16F10) cells. Exogenous HABP1 adhered to the cell surface transiently and was shown to interact and colocalize with α(v)β(3) integrin, a regulatory molecule of cell migration. In HABP1 treated cells, the phosphorylation of nuclear factor inducing kinase (NIK) and IκBα was observed, followed by nuclear translocation of p65 subunit of NFκB, along with its DNA-binding and transactivation, resulting in upregulation of MT1-MMP expression and finally MMP-2 activation. To substantiate our findings, prior to HABP1 treatment, the expression of NIK was reduced by small interfering RNA mediated knockdown and confirmed the inhibition of nuclear translocation of p65 subunit of NFκB and upregulation of MT1-MMP expression. In addition, the use of curcumin, an anti-cancer drug, or GRGDSP, the blocking peptide along with exogenous HABP1, inhibited such NFκB-dependent pathway, confirming that HABP1-induced cell migration is α(v)β(3) integrin-mediated and downstream signaling by NFκB. Finally, we translated the in vitro data in mice model and observed enhanced tumor growth with higher MT1-MMP expression and MMP-2 activation in the tumors upon injection of HABP1 treated melanoma cells. The treatment of curcumin, the anticancer drug along with HABP1, inhibited the migration, expression of MT1-MMP and activation of MMP-2 and finally tumor growth supports the involvement of HABP1 in tumor formation.
细胞迁移是癌症的标志，它调节肿瘤细胞的锚定非依赖性生长和侵袭性。细胞外基质多糖透明质酸（HA）被证明可以调节这个过程。在本报告中，我们证明了从人成纤维细胞 cDNA 中补充纯化的重组透明质酸结合蛋白 1（HABP1/p32/gC1qR）可增强高度侵袭性黑色素瘤（B16F10）细胞的迁移潜力。外源性 HABP1 短暂地附着在细胞表面，并显示与细胞迁移的调节分子α(v)β(3)整合素相互作用和共定位。在 HABP1 处理的细胞中，观察到核因子诱导激酶（NIK）和 IκBα 的磷酸化，随后 p65 亚基 NFκB 核转位，以及其 DNA 结合和转录激活，导致 MT1-MMP 表达上调，最终 MMP-2 激活。为了证实我们的发现，在 HABP1 处理之前，通过小干扰 RNA 介导的敲低降低了 NIK 的表达，并证实了 p65 亚基 NFκB 的核转位和 MT1-MMP 表达的上调受到抑制。此外，使用姜黄素（一种抗癌药物）或 GRGDSP（外源性 HABP1 的阻断肽）抑制了这种 NFκB 依赖性途径，证实了 HABP1 诱导的细胞迁移是由α(v)β(3)整合素介导的，下游信号由 NFκB 介导。最后，我们将体外数据转化为小鼠模型，并观察到在注射 HABP1 处理的黑色素瘤细胞后，肿瘤中 MT1-MMP 表达和 MMP-2 激活增强，肿瘤生长加快。姜黄素（一种抗癌药物）与 HABP1 联合治疗抑制了迁移、MT1-MMP 的表达和 MMP-2 的激活，最终抑制了肿瘤生长，这支持了 HABP1 参与肿瘤形成。
