[肝细胞生长因子对卵巢癌细胞系SKOV3侵袭及其信号转导通路的影响]
[Effect of hepatocyte growth factor on invasion of ovarian cancer cell line SKOV3 and its signal transduction pathway].
作者信息
Wang Hua-Li, Zhang Shu-Lan, Lu Yan-Ming, Jiang Ji-Yong, Zhu Xiao-Yu
机构信息
Department of Obstetrics and Gynecology, The Second Affiliated Hospital, China Medical University, Shenyang, Liaoning, 110004, P. R. China.
出版信息
Ai Zheng. 2006 May;25(5):570-5.
BACKGROUND & OBJECTIVE: Hepatocyte growth factor (HGF) specifically binds to its receptor c-met, activates a complex set of intracellular pathways, and plays important roles in regulating tumor invasion, metastasis, and angiogenesis. HGF and c-met are overexpressed in ovarian cancer. This study was designed to investigate the role of HGF in ovarian cancer invasion and its signal transduction pathway.
METHODS
HGF-induced invasion of ovarian cancer cell line SKOV3 was analyzed with Boyden chamber invasion assay. The expression changes of c-met and matrix metalloproteinase-9 (MMP-9) in SKOV3 cells before and after treatment with HGF and nuclear factor kappaB (NF-kappaB) inhibitor pyrrolidine dithiocarbamate (PDTC) were measured by real-time reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and flow cytometry. The expression of NF-kappaB in SKOV3 cells was evaluated by immunocytochemistry and Western blot.
RESULTS
The invasive cells were significantly more in HGF group than in control group and PDTC plus HGF group (343+/-13 vs. 167+/-11 and 241+/-10, P<0.01). HGF promoted the expression of MMP-9 mRNA by 5.66+/-0.10 folds, but had no effect on c-met mRNA expression; PDTC suppressed the HGF-driven expression of MMP-9 mRNA by 2.75+/-0.80 folds. The positive rates of c-met and MMP-9 were significantly higher in HGF-treated cells than in control cells [(96.6+/-2.0)% vs. (73.3+/-2.4)%, P<0.01; (74.6+/-4.4)% vs. (16.0+/-2.9)%, P<0.01]. The protein levels of c-met and MMP-9 were significantly higher in HGF-treated cells than in control cells (2.84+/-0.18 vs. 1.65+/-0.19, P<0.01; 2.94+/-0.13 vs. 0.54+/-0.18, P<0.01). PDTC significantly suppressed the HGF-driven expression of MMP-9 protein: the positive rate and protein level of MMP-9 were (25.8+/-3.7)% and 0.87+/-0.14 (P<0.05). HGF promoted the expression of NF-kappaB protein in a time-dependent manner. The expression peak appeared 1 h after treatment with HGF (from 0.42+/-0.11 to 1.16+/-0.21, P<0.01). PDTC significantly inhibited the HGF-driven expression of NF-kappaB protein (0.38+/-0.12, P<0.01).
CONCLUSION
HGF might stimulate the invasion of SKOV3 cells by up-regulating the expression of MMP-9 via NF-kappaB pathway.
背景与目的
肝细胞生长因子（HGF）特异性结合其受体c-met，激活一系列复杂的细胞内信号通路，在调节肿瘤侵袭、转移及血管生成中发挥重要作用。HGF和c-met在卵巢癌中均呈过表达。本研究旨在探讨HGF在卵巢癌侵袭中的作用及其信号转导通路。
方法
采用Boyden小室侵袭实验分析HGF诱导的卵巢癌细胞系SKOV3的侵袭能力。运用实时逆转录-聚合酶链反应（RT-PCR）、蛋白质印迹法及流式细胞术检测HGF及核因子κB（NF-κB）抑制剂吡咯烷二硫代氨基甲酸盐（PDTC）处理前后SKOV3细胞中c-met和基质金属蛋白酶-9（MMP-9）的表达变化。通过免疫细胞化学和蛋白质印迹法评估SKOV3细胞中NF-κB的表达。
结果
HGF组侵袭细胞数显著多于对照组及PDTC+HGF组（343±13 vs. 167±11和241±10，P<0.01）。HGF使MMP-9 mRNA表达升高5.66±0.10倍，但对c-met mRNA表达无影响；PDTC使HGF驱动的MMP-9 mRNA表达降低2.75±0.80倍。HGF处理组细胞中c-met和MMP-9的阳性率显著高于对照组[（96.6±2.0）% vs. （73.3±2.4）%，P<0.01；（74.6±4.4）% vs. （16.0±2.9）%，P<0.01]。HGF处理组细胞中c-met和MMP-9的蛋白水平显著高于对照组（2.84±0.18 vs. 1.65±0.19，P<0.01；2.94±0.13 vs. 0.54±0.18，P<0.01）。PDTC显著抑制HGF驱动的MMP-9蛋白表达：MMP-9的阳性率和蛋白水平分别为（25.8±3.7）%和0.87±0.14（P<0.05）。HGF以时间依赖性方式促进NF-κB蛋白表达。HGF处理1小时后表达达到峰值（从0.42±0.11升至1.16±0.21，P<0.01）。PDTC显著抑制HGF驱动的NF-κB蛋白表达（0.38±0.12，P<0.01）。
结论
HGF可能通过NF-κB途径上调MMP-9表达，从而促进SKOV3细胞的侵袭。
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