摘要:目的 应用酶标仪检测5-ALA在不同分化程度的人口腔鳞状细胞癌(OSCC)细胞中产生原卟啉IX ( PpIX)的荧光强度,以期指导临床鉴别和诊断OSCC。方法 将2×104个SAS细胞接种到96孔板中,应用酶标仪检测不同浓度的PpIX水溶液荧光强度,制作PpIX浓度的标准曲线,计算判定系数R2。在SAS 、HSC-3和HSC-4细胞中分别加入浓度为0、0.5、1、1.5、2mmol/L的5-ALA,孵育0、2、4、6、8、10小时后应用酶标仪检测细胞荧光强度。结果 酶标仪检测的荧光强度与PpIX含量呈线性相关(判定系数R2=0.9982),5-ALA在OSCC细胞中最佳的代谢浓度为1mmol/L,代谢时间为4小时。并且5-ALA在中分化的SAS细胞中产生的荧光强度显著高于高分化的HSC-3和HSC-4细胞(P< 0.05)。结论 5-ALA在不同分化程度的OSCC细胞中产生不同的荧光强度,可辅助用于临床鉴别诊断OSCC。
Abstract: Objective In order to guide clinical differential and diagnosis of OSCC, the florescence intensities were detected by the microplate reader to test the endogenous protoporphyrin IX which was produced by 5-ALA in varying degrees of differentiation oral squamous cell carcinoma. Methods 2×104 of SAS cells were cultivated in 96-well plates, the fluorescence intensities of different concentrations of PpIX aquenous solution were detected to make the standard curve of PpIX. 5-ALA of 0, 0.5, 1, 1.5, 2mmol/L were added into SAS, HSC-3 and HSC-4 respectively, and then the fluorescence intensities were detected at 0, 2, 4, 6, 8, 10 hours. Results The fluorescence intensities which were detected by microplate reader were linearly related with PpIX content. The best metabolite concentration of 5-ALA was 1 mmol/L in OSCC cells, and the metabolic time was 4 hours. The fluorescence intensity of 5-ALA in the differentiating SAS cells was significantly higher than the differentiated HSC-3 and HSC-4 cells (P< 0.05). Conclusion 5-ALA produces different fluorescence intensity in varying degrees of differentiation OSCC cells. These results can be used in the differential diagnosis of clinical OSCC.