Inhibitory effect of doxycycline on vasculogenic mimicry in retinoblustoma cell line in vitro

( views:, downloads: )
LIU Fu-ling(Department of Ophthalmology, Qingdao Eye Hospital, Shandong Eye Institute, Qingdao 266021, China)
NIU Ying-jun(Department of Ophthalmology, Qingdao Eye Hospital, Shandong Eye Institute, Qingdao 266021, China)
YUAN Chun-yan(Department of Ophthalmology, Qingdao Eye Hospital, Shandong Eye Institute, Qingdao 266021, China)
Journal Title:
Volume 25, Issue 02, 2009
Key Word:
Retinoblastoma/drug therapy;Doxycycline/administration & dosage;Matrix tnetalloproteinase 9;Matrix metalloproteinase 2;Vasculogenic mimicry

Abstract: Objective To investigate the effect of doxycycline on the proliferation and vasculogenic mimicry in retinoblastoma (RB) cell line in vitro. Methods RB cell line were tested for their ability to form perfusable tubular networks in 3-D culture with doxycycline in the concentrations ranging from 5 to 20 mg/L, and CoCl2 was used as chemical hypoxia-inducing reagent to mimic tumor hypoxic microenvironment. The effect of doxycycline on proliferation were detected by MTT assay in vitro, and the effect on tube formation of RB cells were detected by tube-like structure formation assay and PAS staining. The mRNA levels of MMP-2 and MMP-9 at different hypoxic culture and different doxyeycline concentrations were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Results The micrograph showed that RB cells linked each other to form cavity and network tructure in 3-D culture, the number of tubules in doxycycline group were significantly lower than which in the control group in the concentrations ranging from 5 to 20 mg/L (P<0.001). OD of doxycycline group was significantly lower than which in the control group (t=15.320,P<0.01), The proliferation of RB cells had a negative correlation with the concentration of doxycycline (r=-0.924, P<0.01). The levels of MMP-2 and MMP-9 mRNA of RB cells under hypoxia were significantly higher than which in the control group (t= 16. 469,P<0.01). As the concentration of doxycycline increased, the expression of MMP-2 and MMP-9 decreased. The result of double staining also showed that VM, formed by CD34-negative and PAS-positive tumor cells, existed in 12 simples of retinoblastoma. Conclusion RB cells have the capacity of self-metamorphosing and vasculorizing in 3-D culture. Doxyeycline can inhibit their proliferation and vasculogenic mimicry formation in vitro by down-regulating the expression of MMP-2 and MMP-9.

  • [1]Maniotis AJ,Folberg R,Hess A,et al.Vascular channel formation by human melanoma cells in vivo and in vitro:vasculogenic mimicry.Am J Pathol,1999,155:739-752.
  • [2]Bissell MJ:Tumor plasticity allows vaseulogenic mimicry,a novel form of angiogenic switch-a rose by any other name? Am J Pathol,1999,155:675-679.
  • [3]Shirakawa K,WakasugiH,Heike Y,WatanabeI,Yamada S,Saito K,Konishi F.Vasculogenic mimicry and pseudo comedo formation in breast cancer.Int J Cancer,2002,50;189-201.
  • [4]Kazuo Shirakawa,Hitoshi Tsuda,Yuji Heike,et al.Absence of Endothelial Cells,Central Necrosis,and Fibrosis Are Associated with Aggressive Inflammatory Breast Cancer.Cancer Research,2001,61:445-451.
  • [5]赵秀兰,杜静,张诗武,等.肝细胞癌中血管生成拟态的研究.中华肝脏病杂志,2006,14:41-44.
  • [6]Passalidou E,Trivella M,Singh N,Ferguson M,Hu J,Cesario A,Granone P,Nicholson AG,Goldstraw P,Ratcliffe C,Tetlow M,Leigh I,Harris AL,Garter KCand pezzella F.Vascular phenotype in angiogenic and nonangiogenie lung nonsmall cell carcinomas.Br J Cancer,2002,86:244-249.
  • [7]Sharma N,Seftor RE,Seftor EA,Gruman LM,Heidger PMJr,Cohen MB,Lubaroff DM,Hendrix MJ.Prostatic tumor cell plasticity involves cooperative interactions of distinct phenotypic subpopulations:Role in vasculogenic mimicry.Prostate 2002,50:189-201.
  • [8]陈大年,二十一世纪视网膜母细胞瘤研究:希望与挑战.中华眼底病杂志,2007,23:310-313.
  • [9]Carol L,Shields,孙红等.视网膜母细胞瘤的治疗进展.中华眼底病杂志,2004,20:194-197.
  • [10]Hotary K,Allen E,Puntrieri A,et al.Regulation of cell invasion and morphogcncsis in a three-dimen-sionnal type Ⅰ collagenmatrix by membrane-type metalloproteinase 1,2 and 3.J Cell Biol,2000,149:1309-1323.
  • [11]Ray JM,stetler-Stevenson WG.The role of matrix metalloproteases and their inhibitors in tumor invasion,metasis and angiogenesis.Eur Respir J,1994;7:2062.
  • [12]Fang J,Shing Y,Wiederschain D,et al.Matrix metalloproteinase-2 is required for the switch to the angiogenic phenotype in a tumor model.Proc Natl Acad Sci USA.2000,97:3884-3889.
  • [13]Liu J,Xiong W,Baca-Regen L,et al.Mechanism of inhibition of matrix metalloproteinase-2 expression by doxycycline in human aortic smooth muscle cells.J Vasc Surg,2003,38:1376-1383.
  • [14]Sapadin AN,Fleischmajer R.Tetracyelines:nonantibiotic properties and their clinical implications.J Acad Dermatol,2006,54:258-265.
WanfangData CO.,Ltd All Rights Reserved
About WanfangData | Contact US
Healthcare Department, Fuxing Road NO.15, Haidian District Beijing, 100038 P.R.China
Tel:+86-010-58882616 Fax:+86-010-58882615