Down-regulation of FoxM1 inhibits viability and invasion of gallbladder carcinoma cells, partially dependent on inducement of cellular senescence
Authors: Xin-Sen Xu
Kai Qu
Chang Liu
Jie Tao
Yue-Lang Zhang
Rui-Tao Wang
Fan-Di Meng
Ji-Chao Wei
Shun-Bin Dong
Ya-Feng Dong
Lin Wang
Min-Hui Tai
Yan-Zhou Song
Qi-Fei Wu
Journal: World Journal of Gastroenterology (2014)
Discipline: CLINICAL MEDICINE
Keywords: Forkhead box M1; Gallbladder carcinoma; Invasion; Senescence; Viability.
Affiliations: Jie Tao, Xin-Sen Xu, Kai Qu, Rui-Tao Wang, Fan-Di Meng, Ji-Chao Wei, Shun-Bin Dong, Lin Wang, Chang Liu, Department of Hepatobiliary Surgery, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China.

Abstract:  Aim: To investigate the effect of knockdown of Forkhead box M1 (FoxM1) on the proliferation and invasion capacities of human gallbladder carcinoma (GBC)-SD cells. Methods: Four FoxM1 shRNAs were transfected into GBC-SD cells with Lipofectamine 2000 to select the appropriate shRNA for down-regulation of FoxM1. A recombinant lentivirus for shFoxM1 (Lv-shFoxM1), which expresses FoxM1-speciﬁc shRNA, and a negative control carrying green fluorescent protein, which expresses a scrambled RNA, were constructed. After transfection with the recombinant adenovirus and screened with puromycin, RT-PCR and Western blot were utilized to evaluate the inhibition efficiency. Cell viability was evaluated by MTT assay, and cell migration and invasion were assessed using the Transwell system. Cells were suspended in serum-free medium and seeded into Transwell inserts either uncoated (for migration assay) or coated (for invasion assay) with growth factor-reduced Matrigel. To verify the involvement of FoxM1 in the senescence of tumor cells, staining of senescence β-galactosidase (SA β-gal), the widely used biomarker of cellular senescence, was also performed. Results: After successful transfection of four FoxM1 small interfering RNAs (shRNAs) with Lipofectamine 2000, the shF1822 was selected as the most appropriate shRNA according to its obvious inhibitory effect. The recombinant adenovirus was then constructed with the shF1822 and successfully transfected into the GBC-SD cells, resulting in the significant inhibition of FoxM1 expression at both the mRNA and protein levels, compared with the negative control (P &lt; 0.05). After transfection, down-regulation of FoxM1 significantly inhibited cell viability according to the MTT assay (P &lt; 0.05). In addition, Transwell migration and invasion assays also suggested the suppression of invasion ability of the transfected cells. SA β-gal staining showed that down-regulation of FoxM1 could induce more senescent GBC cells (P &lt; 0.05), suggesting the possible involvement of the senescence process of the FoxM1-deficient cells in GBC. Conclusion: FoxM1 is functionally involved in viability of GBC cells, partially dependent on the inducement of cellular senescence, and is a potential target for GBC therapy. 

Retracted: No

PubPeer comments:
 Figure 3 appears to have image duplications. I m including the fluorescence microscopy photos even though they are redundant because I think it is easier to see the duplication with them. ![file]  Figure 3 Representative photograph (u00d7 100) showing recombinant adenovirus transfection efficiency evaluated by fluorescence microscopy (transfected with the negative control, top; transfected with the shF1822, bottom). A, D: Under an ordinary light microscope; B, E: Under a fluorescence microscope , 
 Partial overlap between two images from different papers was detected. ![file] ,