title: Down-regulation of FoxM1 inhibits viability and invasion of gallbladder carcinoma cells, partially dependent on inducement of cellular senescence.
abstract: OBJECTIVE
  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 < 0.05). After transfection, down-regulation of FoxM1 significantly inhibited cell viability according to the MTT assay (P < 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 < 0.05), suggesting the possible involvement of the senescence process of the FoxM1-deficient cells in GBC.
  
  
  CONCLUSIONS
  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.
authors: 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.
authors: Jie
authors: J
authors: Tao
authors: 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.
authors: Xin-Sen
authors: XS
authors: Xu
authors: 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.
authors: Yan-Zhou
authors: YZ
authors: Song
authors: 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.
authors: Kai
authors: K
authors: Qu
authors: 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.
authors: Qi-Fei
authors: QF
authors: Wu
authors: 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.
authors: Rui-Tao
authors: RT
authors: Wang
authors: 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.
authors: Fan-Di
authors: FD
authors: Meng
authors: 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.
authors: Ji-Chao
authors: JC
authors: Wei
authors: 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.
authors: Shun-Bin
authors: SB
authors: Dong
authors: 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.
authors: Yue-Lang
authors: YL
authors: Zhang
authors: 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.
authors: Min-Hui
authors: MH
authors: Tai
authors: 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.
authors: Ya-Feng
authors: YF
authors: Dong
authors: 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.
authors: Lin
authors: L
authors: Wang
authors: 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.
authors: Chang
authors: C
authors: Liu
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D002277:Carcinoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement; D002470:Cell Survival; D016922:Cellular Senescence; D015536:Down-Regulation; D000072278:Forkhead Box Protein M1; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism*; D005706:Gallbladder Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors; D006801:Humans; D009361:Neoplasm Invasiveness; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection
chemical_list: C107585:FOXM1 protein, human; D000072278:Forkhead Box Protein M1; D051858:Forkhead Transcription Factors; D012333:RNA, Messenger
keywords: Forkhead box M1; Gallbladder carcinoma; Invasion; Senescence; Viability