CAJ | 학술논문

目的：观察神经母细胞瘤细胞在被维甲类化合物诱导分化过程中细胞形态、细胞周期及端粒酶基因表达方面的变化，探讨其作用机理及临床意义。方法：查耳酮酸(R9158)及丁羟胺酸(R8605)均为新一代合成维甲类化合物，采用工作浓度10-6mol/L，加入两个神经母细胞瘤细胞系SH-SY-5Y和SK-N-SH的培养基，作为观察组；与观察组同浓度的DMSO分别加入两个细胞系的培养基，作为溶剂对照组。分别观察用药前后细胞形态学变化、流式细胞术检测细胞周期分布及凋亡、原位杂交细胞化学(ISHH)方法检测端粒酶基因(hTR)的表达。结果：1.SH-SY-5Y细胞在药物处理后第5天有明显的神经突起形成，形态类似成熟神经元。SK-N-SH细胞在药物处理后7天内均无明显变化。2.流式细胞分析发现，SH-SY-5Y细胞在药物处理后第5天明显地阻滞于G1期，S期比例显著减少，G2/M期比例基本不变，并有少量细胞凋亡。SK-N-SH细胞在药物处理第3天及第5天均无明显差别，仅第5天有个别细胞凋亡。3.显微镜下观察，SH-SY-5Y细胞于药物作用5天后，hTR阳性细胞显著减少，SK-N-SH细胞则无明显变化。结论：新合成维甲类化合物R9158及R8605诱导维甲酸敏感细胞SH-SY-5Y发生形态学分化，细胞周期阻滞于G1期，端粒酶基因(hTR)表达显著减少，而维甲酸耐受细胞SK-N-SH则无此变化。端粒酶与其他基因相互关联，是维甲类化合物诱导神经母细胞瘤细胞分化过程中重要的调控靶点。
목적：관찰신경모세포류세포재피유갑류화합물유도분화과정중세포형태、세포주기급단립매기인표체방면적변화，탐토기작용궤리급림상의의。방법：사이동산(R9158)급정간알산(R8605)균위신일대합성유갑류화합물，채용공작농도10-6mol/L，가입량개신경모세포류세포계SH-SY-5Y화SK-N-SH적배양기，작위관찰조；여관찰조동농도적DMSO분별가입량개세포계적배양기，작위용제대조조。분별관찰용약전후세포형태학변화、류식세포술검측세포주기분포급조망、원위잡교세포화학(ISHH)방법검측단립매기인(hTR)적표체。결과：1.SH-SY-5Y세포재약물처리후제5천유명현적신경돌기형성，형태유사성숙신경원。SK-N-SH세포재약물처리후7천내균무명현변화。2.류식세포분석발현，SH-SY-5Y세포재약물처리후제5천명현지조체우G1기，S기비례현저감소，G2/M기비례기본불변，병유소량세포조망。SK-N-SH세포재약물처리제3천급제5천균무명현차별，부제5천유개별세포조망。3.현미경하관찰，SH-SY-5Y세포우약물작용5천후，hTR양성세포현저감소，SK-N-SH세포칙무명현변화。결론：신합성유갑류화합물R9158급R8605유도유갑산민감세포SH-SY-5Y발생형태학분화，세포주기조체우G1기，단립매기인(hTR)표체현저감소，이유갑산내수세포SK-N-SH칙무차변화。단립매여기타기인상호관련，시유갑류화합물유도신경모세포류세포분화과정중중요적조공파점。
Objective: The current study was designed to observe and discuss the variation of neuroblastoma cells in morphology, cell cycle and telomerase gene (hTR) expression induced by retinoids. Methods: Two newly-synthesized retinoids, R9158 and R8605, were supplemented into the media of neuroblastoma cells, SH-SY-5Y and SK-N-SH, with the concentration of 10- 6 mol/L as the treatment group. Meanwhile the same supplements of DMSO were as the control group. Pre-and post-supplement cells were observed for morphological changes, cell cycle was analyzed by flow cytometry (FCM) and hTR expression was analyzed by in situ hybridization chemistry (ISHH). Results: 1) The shape of SH-SY-5Y cells changed significantly at the 5th day of R9158 and R8605 treatment, having multiple neurite extensions just as mature neuron. However no shape change of SK-N-SH cells was found in observation for 7 days. 2)For SH-SY-5Y cells, the percentage of Stage G1, S, G2/M differed much between the treatment group and the control group at 5th day when the cell cycle of the treatment group was retarded at Stage G1 and the ratio of Stage decreased significantly. For SK-N-SH cells, however, there was little variation of cell cycle status in the treatment group. 3)The SH-SY-5Y cell numbers of hTR positive expression in the treatment groups decreased significantly at the 5th day. No decrease was found in the R9158 and R8605 groups of SK-N-SH cells in the treatment groups. Conclusions: During the differentiation of RA-sensitive cells induced by retinoids, the telomerase expression was depressed and the cell cycle retarded;whereas the RA-resistant cells changed little in morphology, telomerase expression and cell cycle as well. The telomerase gene associated with other genes would be an important regulated target in the differentiation induced by retinoids.