食管鳞癌细胞KYSE-70的分化诱导对光动力学应答敏感性的抑制  

Photodynamic sensitivity of esophageal cancer KYSE-70 cells is attenuated by alltrans retinoic acid-induced differentiation

作　　者：孙蕾[1] 李懿[1] 石雨[2] 刘喜龙[2] 杨观瑞[1] 赵立群[1] 杨小静[1] 裘一兵[1] 张亚冰[1] 汲翔[3] 康巧珍[2] 汲振余[1] 

机构地区：[1]郑州大学医药科学研究院肿瘤室,河南省郑州市450052 [2]郑州大学生物工程系,河南省郑州市450001 [3]郑州大学基础医学院,河南省郑州市450001

出　　处：《世界华人消化杂志》2011年第26期2709-2716,共8页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30873002;30972707~~

摘　　要：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 impact of all-trans reti- noic acid (ATRA)-induced cell differentiation on photodynamic sensitivity of human esophageal cancer cell line KYSE-70.METHODS: Both well and poorly differentiated KYSE-450 cell lines were used in this study. KYSE-70 differentiation was induced with 1 μmol/L ATRA and evidenced by cell morphology and proliferation. Phototoxicity after photodynamic therapy (PDT, 450 nm) was detected by MTT assay. Apoptosis was measured by flow cytometry, and morphology of apoptotic cells was visualized after Hoechst 33342 staining. RESULTS: Cells after ATRA treatment exhibited increased size, reduced cytoplasmic and nuclear density, and nuclear enlargement. Cell growth was inhibited compared to control cells. After PDT treatment, the survival of well differentiated KYSE-450 cells and ATRA-treated KYSE-70 cells were reduced compared to poorly differentiated KYSE-70 cells. Cell viability differed significantly between ATRA-treated and non-treated KYSE-70 cells after PDT treatment (54.28% ± 3.64% vs 36.23% ± 7.43%, P < 0.001). The percentage of apoptotic cells in ATRA-induced KYSE-70 cells was less than that in non-treated KYSE-70 cells (18.1% vs 33.3%, P < 0.05). CONCLUSION: ATRA-induced cell differentiation decreases photodynamic sensitivity of esophageal cancer KYSE-70 cells possibly by inducing resistance to apoptosis.

关 键 词：食管癌 细胞分化 光动力学疗法 ALA 

分 类 号：R735.1[医药卫生—肿瘤]