CAJ | 학술논문

目的：探讨以磷脂酰肌醇3-激酶相关激酶蛋白家族成员哺乳动物雷帕霉素靶蛋白（ mTOR）为中心的信号通路在克唑替尼（ crizotinib）诱导的棘皮动物微管结合蛋白样蛋白4-间变性淋巴瘤激酶（ EML4-ALK）融合基因阳性的非小细胞肺癌细胞株H2228凋亡中的作用。方法：根据不同的实验目的处理H2228细胞后，荧光定量PCR检测基因状态，MTT法检测细胞抑制率；流式细胞术检测细胞凋亡和细胞周期；Western blotting 检测细胞mTOR信号通路中关键蛋白的表达及活化水平。结果： Crizotinib对H2228细胞有促凋亡作用，呈时间和剂量依赖性，且能使H2228细胞阻滞在G1期。在使用crizotinib处理的凋亡细胞株中发现mTOR活化水平降低，mTOR上、下游关键蛋白的活化水平都呈下降趋势，肺癌细胞株H2228中特殊表达的融合蛋白EML4-ALK变异体3表达量未受影响，但其活化形式p-ALK明显受到抑制。结论：初步证实mTOR信号通路在crizotinib诱导含有EML4-ALK融合基因的肺癌细胞H2228凋亡中有一定作用，为crizotinib的作用机制提供了依据。
목적：탐토이린지선기순3-격매상관격매단백가족성원포유동물뢰파매소파단백（ mTOR）위중심적신호통로재극서체니（ crizotinib）유도적극피동물미관결합단백양단백4-간변성림파류격매（ EML4-ALK）융합기인양성적비소세포폐암세포주H2228조망중적작용。방법：근거불동적실험목적처리H2228세포후，형광정량PCR검측기인상태，MTT법검측세포억제솔；류식세포술검측세포조망화세포주기；Western blotting 검측세포mTOR신호통로중관건단백적표체급활화수평。결과： Crizotinib대H2228세포유촉조망작용，정시간화제량의뢰성，차능사H2228세포조체재G1기。재사용crizotinib처리적조망세포주중발현mTOR활화수평강저，mTOR상、하유관건단백적활화수평도정하강추세，폐암세포주H2228중특수표체적융합단백EML4-ALK변이체3표체량미수영향，단기활화형식p-ALK명현수도억제。결론：초보증실mTOR신호통로재crizotinib유도함유EML4-ALK융합기인적폐암세포H2228조망중유일정작용，위crizotinib적작용궤제제공료의거。
AIM:To investigate the mammalian target of rapamycin ( mTOR) signaling pathway as the center playing a role in the crizotinib-induced apoptosis of non-small cell lung cancer (NSCLC) cell line H2228, which represents positive echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion gene. METHODS:H2228 cells were processed according to different purposes .Fluorescence quantitative PCR is used to ob-serve the gene states .MTT assay is used to detect the cell inhibition rates .The cell apoptosis and cell cycle were analyzed by flow cytometry .The expression and activation levels of the key proteins in the mTOR signaling pathway were determined by Western blotting .RESULTS:Crizotinib promoted the apoptosis of H 2228 cells in a time-and dose-dependent manner . Crizotinib blocked the H2228 cells staying at the G1 phase.In apoptotic H2228 cells processed with crizotinib, the activa-tion level of mTOR was decreased , and the activation levels of the key proteins in upstream and downstream of mTOR path -way were both declined .The expression level of the fusion protein EML 4-ALK variant 3 was not affected , but its active form of p-ALK was significantly suppressed .CONCLUSION:mTOR signaling pathway has a certain relationship with the crizotinib-induced apoptosis of lung cancer cell H 2228, which represents positive EML4-ALK fusion gene.