四甲基吡啶卟啉-光动力疗法对人宫颈永生化上皮H8细胞的作用  

Effects of TMPyP4-PDT on human immortal cervical epithelial H8 cells

作　　者：宗丽菊 张友忠[1] 王颉[1] 吴淑霞[1] 刘洪丽[1] 张璐[2] 

机构地区：[1]山东大学齐鲁医院妇产科,山东济南250012 [2]兰州大学第一临床医学院,甘肃兰州730000

出　　处：《山东大学学报（医学版）》2015年第1期21-26,33,共7页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(81072122)

摘　　要：目的探讨四甲基吡啶卟啉-光动力疗法(TMPy P4-PDT)对人宫颈永生化上皮H8细胞增殖和凋亡的影响。方法显微镜下观察TMPy P4-PDT后H8细胞形态学变化;采用细胞活力试剂盒(CCK-8)检测细胞的增殖活力;Annexin V-FITC/PI双染试剂盒检测细胞凋亡率;免疫细胞化学SABC法检测TMPy P4-PDT对H8细胞p16INK4a蛋白的表达变化;蛋白免疫印迹法(Western blotting)检测人端粒酶逆转录酶(hTERT)、p21蛋白的表达。结果 TMPy P4-PDT可抑制H8细胞的增殖,诱导细胞凋亡,且在一定范围呈TMPy P4剂量依赖性;免疫细胞化学SABC法结果显示,TM Py P4-PDT可降低细胞p16INK4a蛋白的阳性率;Western blotting结果表明,TMPy P4-PDT下调hTERT的表达,上调p21的表达。结论 TM Py P4-PDT可抑制H8细胞的增殖,诱导H8细胞的凋亡,其作用机制可能与下调p16INK4a、hTERT及上调p21的表达有关。Objective To investigate the effects of photosensitizer 5,10,15,20-Tetrakis （1-methylpyridinium-4-yl） porphyrin based-photodynamic therapy （TMPyP4-PDT）on the proliferation and apoptosis of human immortal cervical epithelial H8 cells.Methods The morphological changes were observed by inverted microscope.Proliferative viability was evaluated with Cell Counting Kit-8.Apoptosis was assessed by Annexin V-FITC/PI Apoptosis Detection Kit.The p16INK4a expression was evaluated by immunocytochemistry Strept Avidin-Biotin-enzyme Complex （SABC）method. Human telomerase reverse transcriptase （hTERT）and p21 expression were analyzed by Western blotting.Results TMPyP4-PDT could inhibit H8 cellular proliferative viability and induce apoptosis in a TMPyP4 dose-dependent man-ner.Immunocytochemistry analysis revealed that TMPyP4-PDT could reduce the positive rate of p16INK4a protein in H8 cells.Western blotting results indicated that TMPyP4-PDT could downregulate hTERT protein expression while upregu-late p21 protein in H8 cells.Conclusion TMPyP4-PDT can inhibit H8 cell proliferation and induce apoptosis.The possible mechanism is related to down-regulation of p16INK4a and hTERT,as well as up-regulation of p21 protein.

关 键 词：宫颈肿瘤 H8细胞 四甲基吡啶卟啉-光动力疗法 人端粒酶逆转录酶 P21 蛋白 P16INK4A蛋白 

分 类 号：R454.2[医药卫生—治疗学]