Knockdown of FoxM1 by siRNA interference decreases cell proliferation, induces cell cycle arrest and inhibits cell invasion in MHCC-97H cells in vitro.
OBJECTIVE
To investigate the effects of small interfering RNA (siRNA) knockdown of forkhead box M1 (FoxM1) on the proliferation and invasion capacities of human hepatocellular carcinoma MHCC-97H cells in vitro.


METHODS
The expression levels of FoxM1 in human hepatocellular carcinoma samples, adjacent non-hepatocellular carcinoma liver samples and MHCC-97 cell lines were detected by RT-PCR and Western blotting. FoxM1 siRNA was transfected into MHCC-97H cells with Lipofectamine 2000. Cell growth was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and cell cycle analysis was performed by flow cytometry. Protein expression levels were evaluated by Western blotting. Anchorage-independent growth and the invasive potency of MHCC-97H cells were measured by soft agar colony formation and a transwell cell invasion assay, respectively.


RESULTS
FoxM1 was over-expressed in hepatocellular carcinoma samples compared to adjacent non-hepatocellular carcinoma liver samples. FoxM1 siRNA was successfully transfected into MHCC-97H cells, resulting in the significant inhibition of FoxM1 mRNA and protein expression. Down-regulation of FoxM1 inhibited cell proliferation, caused cell cycle arrest, and decreased invasion of MHCC-97H cells. Compared with control and mock groups, the FoxM1 siRNA transfected cells showed decreased protein expressions of cyclin B1 and cyclin D1, whereas p27 protein expression was increased. Down-regulation of FoxM1 reduced the expression of matrix metalloproteinase-2 (MMP-2) and urokinase plasminogen activator (uPA).


CONCLUSIONS
FoxM1 is functionally involved in hepatocellular carcinoma cell proliferation and invasion and is a potential target for hepatocellular carcinoma therapy.
Department of Hepatobiliary Surgery, the First Affiliated Hospital, Xi'an Jiaotong University, China.
Qi-fei
QF
Wu
Chang
C
Liu
Ming-hui
MH
Tai
Dong
D
Liu
Lei
L
Lei
Rui-tao
RT
Wang
Min
M
Tian
Yi
Y
Lü
D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002453:Cell Cycle; D049109:Cell Proliferation; D000072278:Forkhead Box Protein M1; D051858:Forkhead Transcription Factors / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D009361:Neoplasm Invasiveness / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*
C107585:FOXM1 protein, human; D000072278:Forkhead Box Protein M1; D051858:Forkhead Transcription Factors; D034741:RNA, Small Interfering