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

doi:10.6040/j.issn.1671-7554.0.2014.581

摘要/Abstract

摘要： 目的探讨四甲基吡啶卟啉-光动力疗法(TMPyP4-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TMPyP4-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TMPyP4-PDT对H8细胞p16^INK4a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 TMPyP4-PDT可抑制H8细胞的增殖,诱导细胞凋亡,且在一定范围呈TMPyP4剂量依赖性;免疫细胞化学SABC法结果显示,TMPyP4-PDT可降低细胞p16^INK4a蛋白的阳性率;Western blotting结果表明,TMPyP4-PDT下调 hTERT的表达,上调p21的表达。结论 TMPyP4-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p16^INK4a、hTERT及上调p21的表达有关。

关键词: 四甲基吡啶卟啉-光动力疗法, H8细胞, 人端粒酶逆转录酶, p21蛋白, p16^INK4a蛋白, 宫颈肿瘤

Abstract: 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 p16^INK4a 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 manner. Immunocytochemistry analysis revealed that TMPyP4-PDT could reduce the positive rate of p16^INK4a protein in H8 cells. Western blotting results indicated that TMPyP4-PDT could downregulate hTERT protein expression while upregulate 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 p16^INK4a and hTERT, as well as up-regulation of p21 protein.

Key words: 5, 10, 15, 20-Tetrakis (1-methylpyridinium-4-yl) porphyrin- photodynamic therapy, p21 protein, Cervix neoplasms, H8 cells, Human telomerase reverse transcriptase, p16^INK4a protein

中图分类号:

R454.2

宗丽菊, 张友忠, 王颉, 吴淑霞, 刘洪丽, 张璐. 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[J]. 山东大学学报（医学版）, 2015, 53(1): 21-26.

ZONG Liju, ZHANG Youzhong, WANG Jie, WU Shuxia, LIU Hongli, ZHANG Lu. Effects of TMPyP4-PDT on human immortal cervical epithelial H8 cells[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2015, 53(1): 21-26.

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