Xu Dianshuang, Ke Yiquan, Jiang Xiaodan, Cai Yingqian, Peng Yiru, Li Yingxin
Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
Br J Neurosurg. 2010 Dec;24(6):660-5. doi: 10.3109/02688697.2010.500416. Epub 2010 Aug 13.
This article reports the phototoxicity effects of a novel photosensitiser ZnPcS4-BSA on human U251 glioma cells in vitro. The cellular uptake of ZnPcS4-BSA by U251 glioma cells was quantified by UV-spectra, and the optimal incubation time was determined. Human U251 glioma cells were incubated in ZnPcS4-BSA of various concentrations, and received laser irradiation of different energy densities. Cell survival rates were measured by CCK-8 assay. Flow cytometer was used to detect apoptosis. Expression of vascular endothelial growth factor (VEGF) gene was detected by real-time PCR in U251 cells after photodynamic therapy (PDT), and β-actin was used as an internal standard. The normal U251 cells severed as controls. Results indicate that the uptake of ZnPcS4-BSA by U251 glioma cells reaches maximum after incubation for 4 hours. ZnPcS4-BSA with different concentrations without light irradiation has no significant effects on cell survival rates. Without ZnPcS4-BSA incubation, cell survival rate of high-dose group (400 J/cm(2)) is the lowest, whereas no significant difference has been found between any other two groups. At laser irradiation of 150 J/cm(2), inhibition rates of the cells increase with ZnPcS4-BSA concentration, and half-inhibitory concentration (IC50) is 0.16 μmol/L. Apoptosis rate of the cells after PDT is significantly higher than that of the control group (p < 0.01). The VEGF expression in the cells increases 5.616 times after PDT. The novel ZnPcS4-BSA is a good photosensitiser for PDT towards U251 glioma cells. The ZnPcS4-BSA based PDT can induce effective apoptosis.
本文报道了一种新型光敏剂ZnPcS4-BSA对人U251胶质瘤细胞的体外光毒性作用。通过紫外光谱对U251胶质瘤细胞摄取ZnPcS4-BSA进行定量，并确定最佳孵育时间。将人U251胶质瘤细胞在不同浓度的ZnPcS4-BSA中孵育，并接受不同能量密度的激光照射。采用CCK-8法检测细胞存活率。用流式细胞仪检测细胞凋亡情况。光动力疗法（PDT）后，采用实时PCR检测U251细胞中血管内皮生长因子（VEGF）基因的表达，以β-肌动蛋白作为内参。以正常U251细胞作为对照。结果表明，U251胶质瘤细胞孵育4小时后对ZnPcS4-BSA的摄取量达到最大。不同浓度的ZnPcS4-BSA在无光照时对细胞存活率无显著影响。未孵育ZnPcS4-BSA时，高剂量组（400 J/cm²）的细胞存活率最低，而其他两组之间无显著差异。在150 J/cm²激光照射下，细胞抑制率随ZnPcS4-BSA浓度增加而升高，半数抑制浓度（IC50）为0.16 μmol/L。PDT后细胞凋亡率显著高于对照组（p < 0.01）。PDT后细胞中VEGF表达增加5.616倍。新型ZnPcS4-BSA是一种对U251胶质瘤细胞进行PDT的良好光敏剂。基于ZnPcS4-BSA的PDT可诱导有效的细胞凋亡。
