竹红菌乙素光动力对人舌鳞癌Tca8113细胞杀伤作用的初步研究

Primary study of lethal effects by HypocrellinB with PDT on human tongue carcinoma Tca8113 cells in vitro

目的:本文是用实验的方法,初步探讨中药光敏剂竹红菌乙素(HB)在光动力(PDT)作用下,对体外人舌鳞癌Tca8113细胞的杀伤作用。方法:常规培养、选取对数生长期人舌鳞癌Tca8113细胞,用含有竹红菌乙素(HB)浓度分别为0μM、0.25μM、0.5μ/M、1μM、2μM的RPMI-1640培养液孵育细胞4h后,经470nmLED定制多光源半导体激光光照处理(PDT),能量密度设置分别为0J/CM2、1J/CM2、2J/CM2、3J/CM2、4J/CM2。PDT处理后细胞继续孵育24h后,分别在光学显微镜和荧光显微镜下观察细胞形态学变化,并用甲基噻唑四唑法(MTT法)检测竹红菌乙素(HB)对细胞的抑制作用。并分别在激光照射6h及24h后,用流式细胞仪测定细胞凋亡率。结果:经PDT处理后细胞在光学显微镜及荧光显微镜下均可观察到细胞坏死和凋亡样改变;在一定浓度范围和光照能量密度范围内,竹红菌乙素(HB)在PDT作用下对人舌鳞癌Tca8113细胞生长增殖的抑制和诱导凋亡作用与药物浓度和能量密度成正相关变化。结论:竹红菌乙素(HB)在光动力(PDT)作用下,对人舌鳞癌Tca8113细胞具有显著的杀伤作用,并在一定范围内呈浓度剂量和光剂量正相关依赖性。

Objective： To primarily explore the lethal effects of traditional Chinese drag photosensitizer hypoerellinB with photedynamic therapy （PDT） on human tongue carcinoma Tca8113 cells in vitro. Methods： Logarithmic growing Tca8113 coils were incubated in RPMI - 1640 medium with hypocrellinB for 5 different concentrations （0μM, 0.25gM, 0.5μM,1 μM, 2μM） for 4 hours, then the chosen cells were irradiated with PI）T by 470nm LED multi - lighthouse semiconductor laser. The light enegy density was decemed to 5 ladders （0J/CM^2. 1 J/CM^2, 2 J/CM^2, 3J/CM^2. 4J/CM2 ）. After 24 hours later, the modality of the hypocrellinB- PDT cells was observed by light microscope and fluorescence microscope, and the proliferative inhibition of the Tca8113 cells was detected by methyl thiazelyhetrazol（MTT）. The coils apoptosis rate was determined by flow cytometry after 6 hours and 24 hotws . Results： The necrosis and apoptosis cells were both seen by light microscope and fluoresconco microscope in the PDT cells. Under definite coucontration range and energy density range, the inhibition rate and the induced apoptosis rate of the hypocrellinB- PDT cells were direct ratio correlative changed with the concentration and the energy density. Conclusion： HypocrellinB - PDT has conspicuous lethal effects on human tongue carcinoma Tea8113 cells, which has direct correlation with the concontration range and energy density range under definite extent.