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目的:构建降表达FOXQ1的慢病毒载体,并探索FOXQ1对基底样(basal-like)亚型乳腺癌细胞增殖转移能力的影响。方法:体外合成FOXQ1干扰片段并与PLKO.1-puro连接构建重组慢病毒质粒PLKO.1-sh FOXQ1,构建重组慢病毒颗粒,分别感染MDA-MB-231和BT549细胞。通过RT-qPCR、Western blot检测细胞中FOXQ1表达情况。通过MTT、克隆形成实验观察稳定降表达FOXQ1后细胞增殖能力的变化情况,采用Transwell实验检测细胞迁移侵袭能力的变化,并用RT-qPCR、Western blot检测增殖转移相关基因表达量的变化。结果:利用重组慢病毒颗粒PLKO.1-sh FOXQ1感染MDA-MB-231和BT549细胞,FOXQ1表达量显著降低,导致MDA-MB-231和BT549细胞的增殖以及迁移侵袭能力显著降低。同时增殖相关基因cyclin D1、c-Myc,转移相关基因fibronectin 1、vimentin的表达下调。结论:沉默FOXQ1的表达能够抑制MDA-MB-231和BT549细胞的增殖和转移。
OBJECTIVE: To construct a lentiviral vector that expresses FOXQ1 and explore the effect of FOXQ1 on the proliferation and metastasis of basal-like subtype breast cancer cells. Methods: FOXQ1 interference fragment was synthesized in vitro and inserted into PLKO.1-puro to construct recombinant lentiviral plasmid PLKO.1-sh FOXQ1. The recombinant lentiviral particles were constructed and infected with MDA-MB-231 and BT549 cells respectively. The expression of FOXQ1 in cells was detected by RT-qPCR and Western blot. The changes of cell proliferation were observed by MTT and colony formation assay. The changes of cell migration and invasion were detected by Transwell assay. The expression of proliferation-related genes was detected by RT-qPCR and Western blot. Results: FOXQ1 expression was significantly decreased in MDA-MB-231 and BT549 cells infected with recombinant lentiviral particles PLKO.1-sh FOXQ1, resulting in a significant decrease in the proliferation and migration of MDA-MB-231 and BT549 cells. Meanwhile, the expression of cyclin D1, c-Myc and fibronectin 1 and vimentin were down-regulated. Conclusion: Silencing FOXQ1 expression can inhibit the proliferation and metastasis of MDA-MB-231 and BT549 cells.