Department of Gastroenterology, Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China.
J Cancer Res Clin Oncol. 2010 Nov;136(11):1641-9. doi: 10.1007/s00432-010-0822-y. Epub 2010 Mar 4.
The aim of this study was to investigate the role of Li-cadherin in invasion and metastasis in LoVo cells.
We applied RNA interference mediated downregulation of Li-cadherin expression in LoVo cells. Li-cadherin expression in LoVo cells was examined by semiquantitative polymerase chain reaction, immunofluorescence, western blot, and immunoprecipitation, respectively. Effect of suppression of Li-cadherin expression on cell migration, invasion, and adhesion was detected by wound healing assay, migration assay, invasion assay, and adhesion assay. Expression and activity of MMP-2 and MMP-9 were analyzed by gelatin zymography.
Cell migration, invasion, and adhesion were increased concomitantly with the reduction in Li-cadherin protein expression. Furthermore, downregulation of Li-cadherin expression induced secretion of proMMP-9, active MMP-9 and active MMP-2.
This study suggests that silencing Li-cadherin has positive actions in the processes of LoVo cells invasion and metastasis, and the interactions among MMP-2, MMP-9, and Li-cadherin participate in the multiple steps of invasion and metastasis in LoVo colorectal cancer cells.
本研究旨在探讨 Li-cadherin 在 LoVo 细胞侵袭和转移中的作用。
应用 RNA 干扰技术下调 LoVo 细胞中 Li-cadherin 的表达。分别采用半定量聚合酶链反应、免疫荧光、Western blot 和免疫沉淀检测 LoVo 细胞中 Li-cadherin 的表达。通过划痕愈合试验、迁移试验、侵袭试验和黏附试验检测抑制 Li-cadherin 表达对细胞迁移、侵袭和黏附的影响。通过明胶酶谱法分析 MMP-2 和 MMP-9 的表达和活性。
随着 Li-cadherin 蛋白表达的降低，细胞迁移、侵袭和黏附能力均增加。此外，下调 Li-cadherin 表达可诱导 proMMP-9、活性 MMP-9 和活性 MMP-2 的分泌。
本研究表明，沉默 Li-cadherin 对 LoVo 细胞侵袭和转移过程具有积极作用，MMP-2、MMP-9 和 Li-cadherin 之间的相互作用参与了 LoVo 结直肠癌细胞侵袭和转移的多个步骤。
