CAJ | 학술논문

目的：探究黄连素对乏氧环境中鼻咽癌细胞系CNE-2及裸鼠移植瘤的增敏效应及其分子机制。方法四甲基偶氮唑盐比色法（ MTT法）检测黄连素对CNE-2细胞的增殖抑制作用，克隆形成实验观察5μmol/L黄连素对常氧和乏氧环境中CNE-2细胞放射敏感性的影响。黄连素乏氧下作用24 h后给予8 Gy X射线照射，流式细胞技术检测CNE-2细胞凋亡率。建立鼻咽癌CNE-2细胞裸鼠移植瘤模型，观察黄连素及单次8 Gy照射对移植瘤的抑制作用。 Western blot检测乏氧条件下黄连素对HIF-1α及VEGF蛋白表达的抑制作用。结果黄连素显著抑制CNE-2细胞增殖，且存在时间、浓度依赖效应，24 h药物作用后半数抑制剂量IC50为（14?9±2?2）μmol/L。乏氧环境下，5μmol/L黄连素处理后的克隆形成率较单纯乏氧组下降，其增敏比（ SERD0）为1?27。在常氧环境中克隆形成率较乏氧组显著增加，常氧组和常氧加药组的SERD0分别为1?45和1?74。5和15μmol/L黄连素能在乏氧环境中单独发挥促凋亡作用，与对照组比较差异有统计学意义（ t＝5?01、9?02，P＜0?05），黄连素联合照射后，凋亡率较单纯照射组增加（t＝5?31、9?91，P＜0?05）。体内实验显示，照射给药组肿瘤体积的增长显著延迟于对照组及单纯给药组，肿瘤倍增时间联合组较单纯照射组及对照组差异有统计学意义（t＝2?96、14?52，P＜0?05）。 Western blot显示乏氧条件下HIF-1α、VEGF表达升高，黄连素作用24 h后表达显著下调。结论黄连素对乏氧环境中的CNE-2细胞及裸鼠移植瘤有放射增敏作用，其与下调乏氧环境中HIF-1α及下游因子VEGF的表达有关。
목적：탐구황련소대핍양배경중비인암세포계CNE-2급라서이식류적증민효응급기분자궤제。방법사갑기우담서염비색법（ MTT법）검측황련소대CNE-2세포적증식억제작용，극륭형성실험관찰5μmol/L황련소대상양화핍양배경중CNE-2세포방사민감성적영향。황련소핍양하작용24 h후급여8 Gy X사선조사，류식세포기술검측CNE-2세포조망솔。건립비인암CNE-2세포라서이식류모형，관찰황련소급단차8 Gy조사대이식류적억제작용。 Western blot검측핍양조건하황련소대HIF-1α급VEGF단백표체적억제작용。결과황련소현저억제CNE-2세포증식，차존재시간、농도의뢰효응，24 h약물작용후반수억제제량IC50위（14?9±2?2）μmol/L。핍양배경하，5μmol/L황련소처리후적극륭형성솔교단순핍양조하강，기증민비（ SERD0）위1?27。재상양배경중극륭형성솔교핍양조현저증가，상양조화상양가약조적SERD0분별위1?45화1?74。5화15μmol/L황련소능재핍양배경중단독발휘촉조망작용，여대조조비교차이유통계학의의（ t＝5?01、9?02，P＜0?05），황련소연합조사후，조망솔교단순조사조증가（t＝5?31、9?91，P＜0?05）。체내실험현시，조사급약조종류체적적증장현저연지우대조조급단순급약조，종류배증시간연합조교단순조사조급대조조차이유통계학의의（t＝2?96、14?52，P＜0?05）。 Western blot현시핍양조건하HIF-1α、VEGF표체승고，황련소작용24 h후표체현저하조。결론황련소대핍양배경중적CNE-2세포급라서이식류유방사증민작용，기여하조핍양배경중HIF-1α급하유인자VEGF적표체유관。
Objective To investigate the radiosensitiation effect of berberine on human nasopharyngeal carcinoma ( NPC) in hypoxia condition and explore the underlying mechanisms. Methods MTT assay, clonogenic assay and flow cytometry were performed to analyze cell proliferation, colony formation and apoptosis, respectively. Male nude mice inoculated subcutaneously with CNE-2 cells were used to examine the radiosensitization effect of berberine in vivo. The expressions of HIF-1α and VEGF were assessed by Western blot. Results Berberine efficiently inhibited the proliferation of CNE-2 cells in time-dependent and dose-dependent fashions with an IC50 of ( 14?9 ± 2?2 ) μmol/L. Clonogenic survival assay showed that berberine ( 5 μmol/L ) sensitized CNE-2 cells to ionizing radiation in hypoxia and its SERD0 was 1?27. Under hypoxic condition, berberine alone (5, 15 μmol/L) could induce apoptosis (t=5?01, 9?02,P<0?05) and it further promoted 8 Gy radiation-induced apoptosis (t =5?31, 9?91,P <0?05). Moreover, berberine significantly delayed the tumor growth in the combination group (berberine +irradiation) compared with the mice received irradiation alone or PBS (t =2?96, 14?52, P <0?05). Immunobloting assay showed that berberine inhibited the upregulation of HIF-1α and VEGF induced by hypoxia in CNE-2 cells. Conclusion Berberine confers radiosensitivity on hypoxic NPC in vitro and in vivo, which is probably associated with the downregulation of HIF-1α and VEGF expressions.