REGγ促进人乳腺癌MDA-MB-231细胞株发生上皮-间充质转化的研究

REGγ induces epithelial-mesenchymal transition in MDA-MB-231 cell lines

摘要:

目的：研究REGγ基因促进人乳腺癌细胞株MDA-MB-231发生上皮-间充质转化的生物作用。方法：免疫组化检测乳腺癌组织、与癌旁组织REGγ的表达差异以及转移淋巴结中REGγ的表达水平。Western blot和RT-PCR检测不同人乳腺癌细胞系REGγ的表达差异。构建REGγ基因过表达和干扰载体并将其导入MDA-MB-231细胞中，然后用RT-PCR和细胞免疫荧光分别从转录水平和蛋白质水平检测上皮-间充质转化细胞标记物的表达水平的变化。分别通过划痕实验和Transwell证实REGγ对细胞侵袭迁移能力的影响。结果：免疫组化提示REGγ在乳腺癌组织（阳性表达率为88.57%）和转移淋巴结（阳性表达率为87.5%）中较癌旁组织（阳性表达率为0）明显高表达（ χ2=25.390，P=0.000； χ2=12.440，P=0.000）。MDA-MB-231细胞过表达REGγ后促进细胞发生上皮-间充质转化，使细胞侵袭能力增强（t=22.974，P=0.000）；干扰REGγ后抑制上皮-间充质转化，使细胞侵袭能力减弱（t=12.118，P=0.000）。结论：REGγ促进人乳腺癌MDA-MB-231细胞发生上皮-间充质转化，并促进肿瘤细胞侵袭转移。

Abstract:

Objective：To study the biological role of REGγ gene in promoting epithelial-mesenchymal transition in MDA-MB-231 cell lines. Methods：Immunohistochemistry was used to detect the expression of REGγ in breast cancer tissues and adjacent tissues. Western blot and RT-PCR were used to detect the expression of REGγ in different human breast cancer cell lines. To construct the stable cell lines with overexpression or interference of REGγ gene，and RT-PCR and cellular immunofluorescence were utilized to de－tect the expression level of epithelial-mesenchymal transformed cell marker respectively. The effects of REGγ on cell migration and invasion were confirmed by scratch test and Transwell respectively. Results：Immunohistochemistry suggested that REGγ was signifi－cantly overexpressed in breast cancer（ χ2=25.390，P=0.000； χ2=12.440，P=0.000）. REGγ induced epithelial-mesenchymal transition and improved the cell invasion（t=22.974，P=0.000）. After interfering with REGγ，epithelial-mesenchymal transition，the cell invasion capacity weakened（t=12.118，P=0.000）. Conclusion：REGγ promotes epithelial-mesenchymal transition in human breast cancer MDA-MB-231 cells and promotes invasion and migration of tumor cells.