层粘连蛋白对人视网膜母细胞瘤株基质金属蛋白酶-2和基质金属蛋白酶-9的影响  

The impact of laminin on the expression of MMP-2 and MMP-9 in human retinoblastoma cells

作　　者：龙华[1] 姜发纲[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科,湖北武汉430022

出　　处：《眼视光学杂志》2007年第3期188-191,共4页Chinese Journal of Optometry & Ophthalmology

摘　　要：目的研究层粘连蛋白(laminin,LN)对人视网膜母细胞瘤株(Hxo-Rb44)基质金属蛋白酶-2(matrix metallopro-teinase,MMP-2)和基质金属蛋白酶-9(matrix metalloproteinase,MMP-9)及其mRNA表达的影响。方法在体外培养的人视网膜母细胞瘤细胞株(Hxo-Rb44)内分别加入含0μg/ml、5μg/ml、10μg/ml、20μg/mlLN的无血清RPMI-1640培养基,培养24h后,分别利用免疫组化SP染色法和PT-PCR检测每个实验组Hxo-Rb44表达MMP-2,MMP-9及其mRNA的含量,统计学方法比较不同实验组Hxo-Rb44表达MMP-2,MMP-9及其mRNA的差异。结果视网膜母细胞瘤细胞经LN刺激后,细胞内MMP-2和MMP-9及其mRNA表达均增加,免疫组化结果显示5μg/ml、10μg/ml、20μg/mlLN处理组MMP-2和MMP-9的染色灰度值分别是178.1±11.7和167.0±15.7、147.9±13.4和140.3±12.0、114.4±13.6和104.0±10.6,和对照组相比,差异有显著性(P<0.05),且LN浓度越高,其表达越强。5μ/ml、10!g/ml、20!g/mlLN处理组的MMP-2/β-actin、MMP-9/β-actin灰度值比值分别是0.181±0.016和0.254±0.023、0.201±0.021和0.287±0.020、0.243±0.019和0.320±0.022,与对照组相比,差异均有显著性(P<0.05),且随着LN浓度增高而增高,呈明显的量效关系。结论LN可诱导视网膜母细胞瘤细胞中表达的MMP-2和MMP-9及其mRNA增加,并呈剂量依赖型。提示LN对视网膜母细胞瘤的侵袭和转移可能具有促进作用。Objective To investigate whether laminin （LN） can affect the expression of matrix metalloproteinase-2 （MMP-2）, matrix metalloproteinase-9 （MMP-9） and their mRNA in the human retinoblastoma cell strain. Methods In vitro cultured human retinoblastoma cells （Hxo-Rb44） were incubated for 24 hours in serum-free RPMI- 1640 with 0, 5, 10 and 20 μg/ml of LN. An immunohistoehemistry staining SP method and reverse transcriptase -polymerase chain reaction （PT-PCR） were used to detect the effect of LN on the expression of MMP-2, MMP-9 and their mRNA. The differences in the expression of MMP-2, MMP-9 and their mRNA in each experimental group were analyzed statistically. Results The expression of MMP-2, MMP-9 and their mRNA increased significantly after co-cuhure with LN. Immunohistochemistry results showed that the respective densities of MMP-2 and MMP-9 in 5,10 and 20 μml for the LN treatment groups were 178.1±11.7 and 167.0±15.7,147.9±13.4 and 140.3±12.0,114.4±13.6 and 104.0±10.6, respectively. The differences in the expression of MMP-2 and MMP-9 in the LN treated group increased significantly （P 〈 0.05） compared to the control group, and the higher the concentration of LN, the higher the expression of MMP-2 and MMP-9. The respective density ratios of MMP-2/βactin and MMP-9/β-actin in 5,1 and 20 μg/ml for the LN treated groups were 0.181±0.016 and 0.254± 0.023,0.201±0.021 and 0.287±0.020,and 0.243±0.019 and 0.320±0.022, respectively. The difference in the expression of their mRNA was statistically significant （P 〈 0.05）compared to the control group. There was an obvious concentration-dependent relationship, the higher the concentration of LN, the higher the mRNA expression of MMP-2 and MMP-9. Condusion It was confirmed that LN increases the expression of MMP-2, MMP-9 and their mRNA in retinohlastoma cells in a concentration-dependent manner, which suggested LN might promote the invasion and metastasis of Rib cells.

关 键 词：视网膜母细胞瘤 基质金属蛋白酶类 层粘连蛋白 

分 类 号：R739.7[医药卫生—肿瘤]