You Position: Home > Paper

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

( views:350, downloads:88 )
Author:
No author available
Journal Title:
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
Issue:
2
DOI:
10.3760/cma.j.issn.1005-1015.2009.02.004
Key Word:
视网膜母细胞瘤/药物疗法;多西环素/投药和剂量;基质金属蛋白酶9;基质金属蛋白酶2;血管生成拟态;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.

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 Email:yiyao@wanfangdata.com.cn