CAJ | 학술논문

目的：探讨靛玉红衍生物PHⅡ-7联合肿瘤坏死因子相关凋亡诱导配体（TRAIL）对乳腺癌细胞MCF-7及其耐药株MCF-7／ADR的增殖及调节TRAIL受体表达的相关机制。方法采用MTT法，分别检测PHⅡ-7、TRAIL及低浓度PHⅡ-7联合TRAIL处理MCF-7、MCF-7／ADR细胞的生长抑制率，同时以TRAIL敏感细胞MDA-MB-231为对照，证实上述乳腺癌细胞对TRAIL的敏感性；采用流式细胞术检测细胞凋亡和药物作用后活性氧的产生情况；real time PCR检测PHⅡ-7以及PHⅡ-7和活性氧抑制剂NAC 联用后，乳腺癌细胞TRAIL功能性受体DR4、DR5的表达情况。结果 PHⅡ-7作用24 h 后对 MCF-7及 MCF-7／ADR 的 IC50分别为（4.49±1.55）、（3.44±0.90）μmol · L－1，TRAIL 可诱导MDA-MB-231细胞凋亡，而MCF-7、MCF-7／ADR对TRAIL极不敏感，各浓度组与MDA-MB-231相比差异具有统计学意义（P＜0.05）；低浓度PHⅡ-7联合TRAIL可有效促进TRAIL对MCF-7、MCF-7／ADR的杀伤并诱导凋亡，而两药单用对上述细胞的增殖抑制作用有限；此外，低浓度的PHⅡ-7还对人正常细胞PBMC的毒性很小，即使与TRAIL联用，在该浓度范围下也不会对正常组织细胞产生明显毒性；PHⅡ-7可有效诱导MCF-7及MCF-7／ADR细胞内活性氧的产生，同时上调TRAIL受体DR4、DR5的水平，并呈剂量依赖性。当PHⅡ-7与ROS 抑制剂 NAC 联用时，可有效抑制 PHⅡ-7对DR4、DR5的表达上调作用。结论低浓度的PHⅡ-7可有效增强乳腺癌细胞MCF-7、MCF-7／ADR对TRAIL治疗的敏感性，其机制可能是通过PHⅡ-7升高细胞内的活性氧水平进而上调TRAIL受体DR4、DR5表达而实现的。本研究为今后PHⅡ-7联合TRAIL 治疗方案的临床应用奠定了基础。
목적：탐토전옥홍연생물PHⅡ-7연합종류배사인자상관조망유도배체（TRAIL）대유선암세포MCF-7급기내약주MCF-7／ADR적증식급조절TRAIL수체표체적상관궤제。방법채용MTT법，분별검측PHⅡ-7、TRAIL급저농도PHⅡ-7연합TRAIL처리MCF-7、MCF-7／ADR세포적생장억제솔，동시이TRAIL민감세포MDA-MB-231위대조，증실상술유선암세포대TRAIL적민감성；채용류식세포술검측세포조망화약물작용후활성양적산생정황；real time PCR검측PHⅡ-7이급PHⅡ-7화활성양억제제NAC 련용후，유선암세포TRAIL공능성수체DR4、DR5적표체정황。결과 PHⅡ-7작용24 h 후대 MCF-7급 MCF-7／ADR 적 IC50분별위（4.49±1.55）、（3.44±0.90）μmol · L－1，TRAIL 가유도MDA-MB-231세포조망，이MCF-7、MCF-7／ADR대TRAIL겁불민감，각농도조여MDA-MB-231상비차이구유통계학의의（P＜0.05）；저농도PHⅡ-7연합TRAIL가유효촉진TRAIL대MCF-7、MCF-7／ADR적살상병유도조망，이량약단용대상술세포적증식억제작용유한；차외，저농도적PHⅡ-7환대인정상세포PBMC적독성흔소，즉사여TRAIL련용，재해농도범위하야불회대정상조직세포산생명현독성；PHⅡ-7가유효유도MCF-7급MCF-7／ADR세포내활성양적산생，동시상조TRAIL수체DR4、DR5적수평，병정제량의뢰성。당PHⅡ-7여ROS 억제제 NAC 련용시，가유효억제 PHⅡ-7대DR4、DR5적표체상조작용。결론저농도적PHⅡ-7가유효증강유선암세포MCF-7、MCF-7／ADR대TRAIL치료적민감성，기궤제가능시통과PHⅡ-7승고세포내적활성양수평진이상조TRAIL수체DR4、DR5표체이실현적。본연구위금후PHⅡ-7연합TRAIL 치료방안적림상응용전정료기출。
Aim To investigate the effect of indirubin derivative PHⅡ-7 and TRAIL on proliferation in breast cancer cell MCF-7 and its MDR counterpart MCF-7/ADR and the mechanism.Methods Growth inhibition rate was examined respectively by MTT assay under treatment with TRAIL or PHⅡ-7 or in combination. Cell apoptosis and ROS production were examined by flow cytometry.The change of TRAIL receptors(DR4/DR5 )in mRNA was analysed by realtime PCR.Re-sults IC50 of PHⅡ-7 on MCF-7 and MCF-7/ADR was (4.49 ±1.55 ),(3.44 ±0.90 )μmol · L-1 respec-tively;MDA-MB-231 was TRAIL sensitive cell line, and apparently TRAIL induced apoptosis in MDA-MB-23 1 .Low concentration of PHⅡ-7 in combination with TRAIL could augment TRAIL-induced cytotoxic effect including apoptosis while TRAIL or PHⅡ-7 treatment alone had limited cytotoxity to those cells.Besides, PHⅡ-7 at this concentration had little toxicity to hu-man peripheral blood mononuclear cells even if in com-bination with TRAIL.PHⅡ-7 generated ROS produc-tion inside MCF-7 and MCF-7/ADR cells and up-regu-lated DR4/DR5 expression concentration dependently. Once upon ROS scavenger NAC involved,the effect of TRAIL receptors up-regualtion by expression was abro-gated.Conclusions PHⅡ-7 at low concentration could improve the sensitivities of breast cancer cell MCF-7 and MCF-7/ADR to TRAIL,the mechanism of which may be the ability of ROS production by PHⅡ-7 help up-regulated TRAIL receptor DR4,DR5 .Our re-search set a solid foundation for PHⅡ-7 in combination with TRAIL in future clinical application.