Key Laboratory of Gastroenterology of Zhejiang Province, Hangzhou, China.
Cancer Sci. 2012 Dec;103(12):2082-91. doi: 10.1111/cas.12018. Epub 2012 Oct 30.
Recent studies have shown that overexpression of regenerating gene family member 4 (REG4) is associated with the initiation and progression of pancreatic cancer. In our study, we explored the role of REG4 in the invasion of pancreatic cancer. Real-time PCR and Western blot analysis were used to determine REG4 expression in pancreatic cancer cell lines. An MTT assay was carried out to test the effect of REG4 on the growth of pancreatic cancer cells. The involvement of REG4 in cancer cell invasion was examined by Transwell invasion assay. Two MMPs, MMP-7 and MMP-9, were identified from a pool of candidate genes as being related to REG4-induced cell invasion by PCR and Western blotting. Immunohistochemistry was used to confirm the correlation between REG4 and the two MMPs. High expression of REG4 was found in BXPC-3 cells and its culture media. But in PANC-1 and ASPC-1 cell lines, REG4 expression levels were very low, and no detectable protein was found in the culture medium. The MTT and Transwell invasion assays showed that recombinant REG4 protein and BXPC-3 conditioned media significantly promoted the proliferation and invasiveness of pancreatic cancer cells. It was also shown that MMP-7 and MMP-9 are upregulated by REG4 induction using real-time PCR and Western blotting analysis. Immunohistochemical study further verified this result. In conclusion, REG4 promotes not only growth but also in vitro invasiveness of pancreatic cancer cells by upregulating MMP-7 and MMP-9.
最近的研究表明，再生基因家族成员 4（REG4）的过表达与胰腺癌的发生和发展有关。在我们的研究中，我们探讨了 REG4 在胰腺癌侵袭中的作用。实时 PCR 和 Western blot 分析用于确定胰腺癌细胞系中 REG4 的表达。MTT 测定用于测试 REG4 对胰腺癌细胞生长的影响。通过 Transwell 侵袭测定研究了 REG4 对癌细胞侵袭的影响。从一组候选基因中鉴定出两种 MMP（MMP-7 和 MMP-9），通过 PCR 和 Western blot 证实与 REG4 诱导的细胞侵袭有关。免疫组织化学用于证实 REG4 与两种 MMP 之间的相关性。发现 REG4 在 BXPC-3 细胞及其培养基中高表达。然而，在 PANC-1 和 ASPC-1 细胞系中，REG4 表达水平非常低，培养基中未检测到可检测的蛋白质。MTT 和 Transwell 侵袭测定表明，重组 REG4 蛋白和 BXPC-3 条件培养基显著促进了胰腺癌细胞的增殖和侵袭性。实时 PCR 和 Western blot 分析还表明，REG4 诱导 MMP-7 和 MMP-9 的上调。免疫组织化学研究进一步证实了这一结果。总之，REG4 通过上调 MMP-7 和 MMP-9 不仅促进了胰腺癌细胞的生长，还促进了其体外侵袭性。
