摘要
背景与目的:失巢凋亡抑制因子酪氨酸激酶受体B(tyrosine kinase receptor B,TrkB)能诱导正常上皮细胞的恶性转化并且使该细胞具有高转移能力。TrkB在神经母细胞瘤和其他多种人类高侵袭性恶性肿瘤组织中过度表达,TrkB在卵巢癌细胞中的表达及其与卵巢癌细胞的高侵袭能力的关系的研究鲜有报道。本研究探讨失巢凋亡抑制因子酪氨酸激酶受体B(tyrosine kinase receptor B,TrkB)与人卵巢癌OVCAR-3细胞侵袭能力的关系。方法:RT-PCR和Western印迹法检测卵巢癌细胞系OVCAR-3细胞贴壁培养、细胞立体培养以及细胞立体培养得到的多细胞团簇经胰酶消化成单细胞后再次贴壁培养细胞TrkB的表达差异;体内、体外细胞侵袭实验检测通过RNAi技术沉默TrkB后OVCAR-3细胞侵袭及转移能力的改变。结果:RT-PCR结果表明,OVCAR-3多细胞团簇中TrkBmRNA的表达高于贴壁细胞(P<0.001);Western印迹结果显示,与OVCAR-3贴壁细胞比较,立体培养细胞中全长TrkB表达升高[(17.47±0.58)%vs(8.93±0.19)%,(P<0.001)]。体内、体外细胞侵袭实验表明,OVCAR-3细胞团簇的侵袭转移能力较普通贴壁细胞明显增强(P<0.01);TrkB沉默后OVCAR-3的侵袭及转移能力受到抑制(P<0.01)。结论:TrkB通过失巢凋亡抑制机制介导了卵巢上皮性癌细胞的侵袭和转移。
Background and purpose：TrkB, a neurotrophic tyrosine kinase receptor,serves as a potent and specific suppressor of anoikis of non-malignant epithelial cells, and confers high metastatic capacity to these cells. TrkB over- expression has been found in neuroblastoma and some other high aggressive cancers. However, the clinical and biologyical signifi cance of the expression of TrkB in human ovarian cancer were rarely reported in literature.TrkB（tyrosine kinase receptor B） is an anoikis suppressor. This study investigated the relationship between TrkB（tyrosine kinase receptor B） expression and invasion capacity in OVCAR-3 ovarian cancer cells. Methods：The expression of TrkB in OVCAR-3 ovarian cancer cells under different culture conditions was evaluated by RT-PCR and Western blotting. The difference of invasion and metastatic ability of OVCAR-3 cells cultured was investigated both in vitro and in vivo by Matrigel invasion assay with or without TrkB silenced by RNAi. Results：The expression of TrkB mRNA in multicellular spheroids was signifi cantly higher than that in OVCAR-3 cells growing in monolayer adhesive culture （P〈0.001）. Compared with OVCAR-3 cells in monolayer adhesive culture, full-length TrkB was overexpressed in OVCAR-3 multicellular spheroids （in anchorage -independent culture） （P〈0.001） detected by Western blotting, and OVCAR3 multicellular spheroids has higher invasion ability than OVCAR3 cells under monolayer adhesive culture, which penetrating cells of the two groups were 71.83±0.75 and 47.67±0.82 （P〈0.01）.The metastatic ability of OVCAR-3 cells was attenuated when TrkB was silenced, the penetrating cells was 37.8±0.8 （P〈0.01）. Conclusion：As a anoikis-suppressor, TrkB may mediate the tumor cells invasion and metastasis in human ovarian epithelial cancer.
出处
《中国癌症杂志》
CAS
CSCD
2008年第10期728-733,共6页
China Oncology
基金
国家自然科学基金资助项目(No:30371482)
上海市卫生局重点研究项目(No:2005ZD002)。
