长春新碱对视网膜母细胞瘤细胞整合素连接激酶表达的抑制作用

Inhibition of vincristine on expression of integrin-linked kinase in retinoblastoma cells

目的探讨视网膜母细胞瘤(Rb)细胞中整合素连接激酶(integrin-linked kinase,ILK)的表达以及长春新碱对ILK表达的影响。方法采用免疫组织化学检测9例Rb组织中ILK的表达。分别在0mg·mL-1、10mg·mL-1、20mg·mL-1、40mg·mL-1、80mg·mL-1长春新碱浓度条件下体外培养Rb细胞48h,采用逆转录聚合酶链反应(RT-PCR)、荧光定量PCR、Western blotting检测不同浓度组ILK mRNA和蛋白的表达。结果 9例Rb组织中ILK的表达呈阳性。体外细胞实验显示:与对照组相比,培养48h后出现长春新碱抑制人Rb细胞ILK mRNA表达,并呈浓度依赖性,在20mg·mL-1组其抑制作用达到高峰,荧光定量PCR检测发现,20mg·mL-1组ILK/18S值明显较对照组低(F=228.130,P<0.05),Westernblotting检测结果与RT-PCR、荧光定量PCR趋势相同。结论 ILK在视网膜母细胞瘤组织中有表达;长春新碱可抑制ILK mR-NA和蛋白的表达。

Objective To investigate expression of integrin-linked kinase（ILK）in retinoblastoma（Rb）cells and effects of vincristine on expression of ILK.Methods ILK expression in 9 Rb cases was examined by immunohistochemistry.Rb cells were cultured with 0 mg·L-1,10 mg·L-1,20 mg·L-1,40 mg·L-1 and 80 mg·L-1 vincristine for 48 hours in vitro.Expression of mRNA and protein of ILK in different groups were detected by RT-PCR,fluorescent quantification PCR and Western blotting.Results Expression of ILK was positive in all 9 cases.In vitro experiment showed：Compared with control group,vincristine could inhibit expression of ILK mRNA in human Rb cells after culturing for 48 hours,and with dose-dependent manner;Its inhibitive effect went to peak at 20 mg·L-1.Fluorescence quantification PCR found that ILK/18S value in 20 mg·L-1group was obviously lower than that in control group（F=228.130,P0.05）.Results of Western blotting were consistent with those of RT-PCR or fluorescence quantification PCR.Conclusions Expression of ILK can be found in Rb cells.Vincristine can decrease the expression of ILK mRNA and protein.