Sun Hua, Liu Geng-Tao
Department of Pharmacology, Institute of Materia Medica, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing, 100050, P. R. China.
Ai Zheng. 2006 Dec;25(12):1464-9.
BACKGROUND & OBJECTIVE: Invasion and metastasis is the main cause of the poor prognosis and high mortality of hepatocellular carcinoma (HCC). Inhibiting invasion and metastasis is an effective approach to reduce the development and mortality of HCC. Dimethyl dicarboxylate biphenyl (DDB) is an old anti-hepatitis drug used in clinics. This study was to assess the inhibitory effect of DDB on adhesion and invasion of HCC cell line MHCC97-H with high metastasis potential, and explore its possible mechanism.
MTT assay was used to evaluate the cytotoxicity of DDB to MHCC97-H cells and the inhibitory effect of DDB on the adhesion of MHCC97-H cells to laminin (LN) and fibronectin (FN). The inhibitory effect of DDB on the invasion of MHCC97-H cells was detected by transwell chamber experiment. The mRNA levels of vascular endothelial growth factor (VEGF), nm23-H1, and urokinase-type plasminogen activator receptor (uPAR) were determined by reverse transcription-polymerase chain reaction (RT-PCR). The secretion and expression of alpha fetoprotein (AFP) were analyzed by ELISA and flow cytometry, respectively.
DDB at non-cytotoxic concentrations (10, 50 and 100 micromol/L) obviously inhibited the adhesion of MHCC97-H cells to LN and FN. The inhibition rates of the invasion of MHCC97-H cells were 25.8% and 32.3% after treatment of 50 and 100 micromol/L DDB, respectively; 50 and 100 micromol/L DDB also decreased the mRNA levels of VEGF, nm23-H1, and uPAR in MHCC97-H cells. The inhibition rates of AFP secretion of MHCC97-H cells were 16.5%, 17.5%, and 48.5% after treatment of 50, 100, and 200 micromol/L DDB, respectively; it also decreased the expression of AFP in MHCC97-H cells.
DDB, at non-cytotoxic concentrations, may obviously suppress the invasion of MHCC97-H cells through inhibiting VEGF, nm23-H1, and uPAR expression.
侵袭和转移是肝细胞癌（HCC）预后差和死亡率高的主要原因。抑制侵袭和转移是降低HCC进展和死亡率的有效方法。联苯双酯是一种临床使用的老抗肝炎药。本研究旨在评估联苯双酯对具有高转移潜能的HCC细胞系MHCC97-H黏附与侵袭的抑制作用，并探讨其可能机制。
采用MTT法评估联苯双酯对MHCC97-H细胞的细胞毒性以及对MHCC97-H细胞与层粘连蛋白（LN）和纤连蛋白（FN）黏附的抑制作用。通过Transwell小室实验检测联苯双酯对MHCC97-H细胞侵袭的抑制作用。采用逆转录-聚合酶链反应（RT-PCR）测定血管内皮生长因子（VEGF）、nm23-H1和尿激酶型纤溶酶原激活物受体（uPAR）的mRNA水平。分别通过ELISA和流式细胞术分析甲胎蛋白（AFP）的分泌和表达。
非细胞毒性浓度（10、50和100 μmol/L）的联苯双酯明显抑制MHCC97-H细胞与LN和FN的黏附。50和100 μmol/L联苯双酯处理后，MHCC97-H细胞侵袭的抑制率分别为25.8%和32.3%；50和100 μmol/L联苯双酯还降低了MHCC97-H细胞中VEGF、nm23-H1和uPAR的mRNA水平。50、100和200 μmol/L联苯双酯处理后，MHCC97-H细胞AFP分泌的抑制率分别为16.5%、17.5%和48.5%；它还降低了MHCC97-H细胞中AFP的表达。
非细胞毒性浓度的联苯双酯可能通过抑制VEGF、nm23-H1和uPAR表达明显抑制MHCC97-H细胞的侵袭。
