Zhou Lei, Zhang Ping-hai, Xu Xin, Xu Nuo, Gao Lei, Bai Chun-xue, Zhang Xin
Department of Respiratory Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Zhonghua Yi Xue Za Zhi. 2009 Sep 15;89(34):2424-8.
To investigate the change of proliferation and invasiveness of non-small cell lung cancer (NSCLC) cell lines SPC-A-1, A549 and LTEP-a-2 with forkhead box M1 (FoxM1) expression deficiency.
A siRNA targeting FoxM1 was designed to deplete the FoxM1 expression of these cell lines and an unrelated siRNA used as control. Real-time RT-PCR and Western blotting were used to examine the FoxM1 expression in mRNA and protein level respectively. Colony assay, wound healing assay and transwell chamber assay were employed to evaluate the colony formation ability and invasiveness of FoxM1 deficient cells.
The designed siRNA could efficiently deplete FoxM1 expression by 83.9%, 83.6% and 88.6% in SPC-A-1, A549 and LTEP-a-2 cell lines respectively. Real-time RT-PCR and Western blot test showed that the FoxM1 protein was also depleted. The colony formation numbers (136.0 +/- 15.5, 87.0 +/- 2.6 and 121.7 +/- 9.4 respectively) and invasion cell numbers (19.2 +/- 2.5, 4.2 +/- 0.8 and 6.2 +/- 1.8 respectively) of FoxM1 deficient SPC-A-1, A549 and LTEP-a-2 cell lines were significantly fewer than those of the unrelated-siRNA transfected group (colony formation numbers were 222.3 +/- 20.5, 164.7 +/- 14.1 and 260.7 +/- 13.5 respectively, and invasive cell numbers were 81.4 +/- 6.2, 39.2 +/- 4.6 and 35.6 +/- 3.0 respectively, all P < 0.01). The cell migration rate of siFoxM1 deficient SPC-A-1 (52.6% +/- 7.8%) was significantly lower than that of the unrelated-siRNA transfected group (85.3% +/- 18.6%, P < 0.01).
The proliferation and invasiveness of several NSCLC cell lines were significantly inhibited after the FoxM1 gene expression was depleted. It suggests that inhibiting the FoxM1 expression might be a promising way for lung cancer therapy.
研究叉头框M1（FoxM1）表达缺失对非小细胞肺癌（NSCLC）细胞系SPC-A-1、A549和LTEP-a-2增殖及侵袭能力的影响。
设计针对FoxM1的小干扰RNA（siRNA）以降低这些细胞系中FoxM1的表达，并用无关siRNA作为对照。分别采用实时逆转录聚合酶链反应（Real-time RT-PCR）和蛋白质免疫印迹法检测FoxM1在mRNA和蛋白水平的表达。采用集落形成实验、伤口愈合实验和Transwell小室实验评估FoxM1缺失细胞的集落形成能力和侵袭能力。
设计的siRNA可分别有效降低SPC-A-1、A549和LTEP-a-2细胞系中FoxM1的表达，降低幅度分别为83.9%、83.6%和88.6%。Real-time RT-PCR和蛋白质免疫印迹检测显示FoxM1蛋白也减少。FoxM1缺失的SPC-A-1、A549和LTEP-a-2细胞系的集落形成数（分别为136.0±15.5、87.0±2.6和121.7±9.4）和侵袭细胞数（分别为19.2±2.5、4.2±0.8和6.2±1.8）明显少于转染无关siRNA组（集落形成数分别为222.3±20.5、164.7±14.1和260.7±13.5，侵袭细胞数分别为81.4±6.2、39.2±4.6和35.6±3.0，均P<0.01）。siFoxM1缺失的SPC-A-1细胞迁移率（52.6%±7.8%）明显低于转染无关siRNA组（85.3%±18.6%，P<0.01）。
FoxM1基因表达缺失后，几种NSCLC细胞系的增殖和侵袭能力受到明显抑制。提示抑制FoxM1表达可能是一种有前景的肺癌治疗方法。
