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目的:研究全反式维甲酸(ATRA)对低分化食管鳞癌细胞系KYSE-70的分化诱导作用及对光动力学疗法(photodynamic therapy,PDT)的应答敏感程度.方法:以高分化食管鳞癌细胞KYSE-450和低分化食管鳞癌细胞KYSE-70为研究对象,用1μmol/LATRA为诱导剂,诱导KYSE-70细胞从低分化状态向高分化状态分化,通过细胞形态学、增殖实验来验证;细胞以1mmol/LALA处理,不同剂量的450nm蓝光照射,MTT法测定PDT对细胞的光毒毒性;流式细胞法测定PDT诱导的凋亡水平,Hoechst33342染色后观察凋亡细胞的胞核形态.结果:经ATRA处理后,诱导组与对照组相比,细胞变扁平、体积增大、胞质密度减低、核变大、核密度亦减低、细胞生长缓慢.高分化KYSE-450、分化诱导后的KYSE-70细胞和未诱导KYSE-70细胞用ALA处理后进行蓝光PDT,MTT结果显示高分化KYSE-450和分化诱导后KYSE-70的细胞存活率明显高于未诱导细胞,而高分化KYSE-450细胞敏感性略微低于分化诱导后的KYSE-70细胞.当光剂量为225mJ/cm2时,诱导前后细胞存活率分别为36.23%±7.43%和54.28%±3.64%,有极显著差异(P<0.001);分化诱导后的KYSE-70细胞凋亡率(18.1%)亦低于未诱导的细胞(33.3%).结论:经分化诱导后的食管鳞癌细胞PDT敏感性明显差于未诱导的食管鳞癌细胞,提示细胞分化诱导疗法不仅不能增强PDT效应,反而降低疗效;细胞分化诱导对PDT效果的影响部分通过抑制凋亡而实现.
AIM: To investigate the differentiation-inducing effect of all-trans retinoic acid (ATRA) on poorly differentiated esophageal squamous cell carcinoma cell line KYSE-70 and the sensitivity to photodynamic therapy (PDT) .Methods: KYSE-450 cells and KYSE-70 cells were used to induce differentiation of KYSE-70 cells from well-differentiated to well-differentiated cells by using 1μmol / L ATRA as an inducer. Cell morphological and proliferation experiments The cells were treated with 1mmol / LALA and different doses of 450nm blue light. The phototoxicity of PDT was measured by MTT assay. The apoptosis induced by PDT was measured by flow cytometry. The morphology of apoptotic cells was observed by Hoechst33342 staining Results: After treatment with ATRA, the cells in the induction group became flattened and enlarged in volume, the cytoplasm density was reduced, the nuclei became larger, the nuclear density was also decreased and the cell growth was slowed down.Compared with the control group, the differentiation of KYSE-450 After KYSE-70 cells and KYSE-70 cells were treated with ALA, blue light PDT was performed. MTT results showed that the cell survival rate of KYSE-450 cells after differentiation and KYSE-70 cells differentiation was significantly higher than that of non-induced cells, KYSE-450 cells The sensitivity was slightly lower than that of differentiation-induced KYSE-70 cells.When the light dose was 225mJ / cm2, the cell viability was 36.23% ± 7.43% and 54.28% ± 3.64%, respectively, with significant difference (P <0.001) ; The apoptosis rate of KYSE-70 cells induced by differentiation was also lower than that of non-induced cells (33.3%). Conclusion: PDT sensitivity of differentiated esophageal squamous carcinoma cells was significantly lower than that of non-induced esophageal squamous cell carcinoma Cancer cells, suggesting that induction of cell differentiation therapy can not only enhance the PDT effect, but reduce the effect; induced by cell differentiation on the PDT effect by inhibiting apoptosis in part.