Department of Hepatobiliary Surgery, the First Affiliated Hospital, Xi'an Jiaotong University, China.
Acta Pharmacol Sin. 2010 Mar;31(3):361-6. doi: 10.1038/aps.2010.4. Epub 2010 Feb 15.
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.
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.
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).
FoxM1 is functionally involved in hepatocellular carcinoma cell proliferation and invasion and is a potential target for hepatocellular carcinoma therapy.
研究小干扰 RNA（siRNA）敲低叉头框 M1（FoxM1）对体外人肝癌 MHCC-97H 细胞增殖和侵袭能力的影响。
采用 RT-PCR 和 Western blot 检测 FoxM1 在人肝癌组织标本、癌旁非肝癌肝组织标本和 MHCC-97 细胞系中的表达水平。用 Lipofectamine 2000 将 FoxM1 siRNA 转染至 MHCC-97H 细胞。用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）比色法评估细胞生长情况，用流式细胞术进行细胞周期分析。用 Western blot 评估蛋白表达水平。用软琼脂集落形成实验和 Transwell 细胞侵袭实验分别测量 MHCC-97H 细胞的无锚定生长和侵袭能力。
FoxM1 在肝癌组织标本中表达高于癌旁非肝癌肝组织标本。FoxM1 siRNA 成功转染至 MHCC-97H 细胞，导致 FoxM1 mRNA 和蛋白表达显著抑制。下调 FoxM1 抑制细胞增殖，引起细胞周期停滞，并降低 MHCC-97H 细胞的侵袭能力。与对照组和空载组相比，FoxM1 siRNA 转染细胞的细胞周期蛋白 B1 和 D1 蛋白表达减少，而 p27 蛋白表达增加。下调 FoxM1 降低了基质金属蛋白酶-2（MMP-2）和尿激酶纤溶酶原激活物（uPA）的表达。
FoxM1 参与肝癌细胞增殖和侵袭，是肝癌治疗的潜在靶点。
