CAJ | 학술논문

目的：探讨唑来膦酸（zoledronic acid，ZOL）及联合紫杉醇（paclitaxel，PTX）体外对人肺腺癌A-549细胞增殖影响及潜在机制。方法：实验组包括不同浓度ZOL组、2 nM PTX组与ZOL联合应用PTX加药顺序不同3个组。MTT方法观测不同浓度ZOL与联合PTX对A-549细胞增殖的影响；流式细胞仪检测各组细胞凋亡；RT-PCR检测各组ERK和AKT mRNA的变化；West-ern blot检测各组ERK、AKT蛋白表达及磷酸化水平和Bcl-2的表达。结果：ZOL能抑制A-549细胞生长，该作用随ZOL浓度增加而增强；ZOL与PTX具有协同抗肿瘤作用，作用大小与两药加药顺序有关，产生最大抑制作用的顺序为先PTX后ZOL；RT-PCR结果显示不同组ERK和AKT mRNA表达水平分别为空白对照组最高，单药ZOL组次之，再次为PTX组，先PTX后ZOL组最低；West-ern blot结果显示各实验组中ERK、AKT蛋白表达水平无差异，但ERK和AKT磷酸化水平有差异，分别为空白对照组最高，单药ZOL组次之，再次为PTX组，先PTX后ZOL组最低，Bcl-2的表达水平亦是如此。结论：ZOL联合PTX可能通过下调RAF/MEK/ERK信号通路中ERK基因mRNA的表达，降低ERK磷酸化水平，抑制ERK激酶活性，进而抑制A-549细胞生长和增殖；同时可能通过下调PI3K/AKT信号通路中AKT基因mRNA的表达，降低AKT磷酸化水平，进而减弱AKT活性，进一步降低抗凋亡蛋白Bcl-2水平，从而起到促凋亡作用。
목적：탐토서래련산（zoledronic acid，ZOL）급연합자삼순（paclitaxel，PTX）체외대인폐선암A-549세포증식영향급잠재궤제。방법：실험조포괄불동농도ZOL조、2 nM PTX조여ZOL연합응용PTX가약순서불동3개조。MTT방법관측불동농도ZOL여연합PTX대A-549세포증식적영향；류식세포의검측각조세포조망；RT-PCR검측각조ERK화AKT mRNA적변화；West-ern blot검측각조ERK、AKT단백표체급린산화수평화Bcl-2적표체。결과：ZOL능억제A-549세포생장，해작용수ZOL농도증가이증강；ZOL여PTX구유협동항종류작용，작용대소여량약가약순서유관，산생최대억제작용적순서위선PTX후ZOL；RT-PCR결과현시불동조ERK화AKT mRNA표체수평분별위공백대조조최고，단약ZOL조차지，재차위PTX조，선PTX후ZOL조최저；West-ern blot결과현시각실험조중ERK、AKT단백표체수평무차이，단ERK화AKT린산화수평유차이，분별위공백대조조최고，단약ZOL조차지，재차위PTX조，선PTX후ZOL조최저，Bcl-2적표체수평역시여차。결론：ZOL연합PTX가능통과하조RAF/MEK/ERK신호통로중ERK기인mRNA적표체，강저ERK린산화수평，억제ERK격매활성，진이억제A-549세포생장화증식；동시가능통과하조PI3K/AKT신호통로중AKT기인mRNA적표체，강저AKT린산화수평，진이감약AKT활성，진일보강저항조망단백Bcl-2수평，종이기도촉조망작용。
Objective:This study aimed to investigate the inhibitory effect of zoledronic acid (ZOL) alone or the combined treat-ment of ZOL and paclitaxel (PTX) on the cell growth of lung cancer cell line in vitro. Methods:The effects of different concentrations of ZOL alone, 2 nM PTX, and combined treatment of ZOL and PTX on the growth of A-549 cell line were determined using methyl thi-azolyl tetrazolium (MTT) method. The mechanism of the curative effect was analyzed by flow cytometry on the basis of apoptotic rate. AKT, phospho-AKT, ERK, phospho-ERK, and Bcl-2 expressions were determined by western blot analysis. AKT and ERK gene ex-pressions were determined by RT-PCR. Results:MTT results showed that ZOL alone could inhibit the growth of lung cancer cell line A-549 in a dose-dependent manner. The combined therapy of ZOL and PTX could inhibit cell growth. This combined treatment is more effective than the single treatment with either ZOL or PTX alone. The synergistic inhibition rate is dependent on drug sequencing. Fur-thermore, maximum inhibition was induced by sequence order, i.e., initial treatment with PTX and then with ZOL. RT-PCR results dem-onstrated that the mRNA of ERK and AKT of the group treated with PTX and then ZOL were lower than that in other treatment groups. Western blot analysis results demonstrated that the ERK and AKT levels of the treated groups were parallel in the cell line. However, the lowest phospho-ERK, phospho-AKT, and Bcl-2 levels were observed in the PTX then ZOL group. The cell lines treated with PTX alone and ZOL alone ranked second and third among the lowest results, respectively. The highest level was observed in the control group. Conclusion: The combined ZOL and PTX treatment induced the downregulation of phospho-ERK, phospho-AKT, and Bcl-2 protein expressions in RAF/MEK/ERK and PI3K/AKT signaling pathway. This pathway could be one of the synergistic antitumor mechanisms of the two drugs.