CAJ | 학술논문

目的探讨紫杉醇诱导白血病细胞凋亡的信号途径及死亡结构域沉默子(SODD)分子靶向对紫杉醇化疗的增敏作用.方法采用Western blot法检测紫杉醇对人急性淋巴细胞白血病Jurkat细胞SODD、B细胞淋巴瘤/白血病-2基因(Bcl-2)、含半胱氨酸的天冬氨酸蛋白水解酶(Caspase-3)、核转录因子(NF-κB)-P65蛋白表达的影响;构建1个载体编码3条质粒表达载体的SODD小分子干扰质粒,转染至Jurkat细胞,采用碘化丙啶(PI)标记流式细胞术检测紫杉醇对细胞的凋亡诱导率.结果紫杉醇在诱导Jurkat白血病细胞凋亡过程中能显著下调SODD及Bcl-2蛋白的表达,在该凋亡过程中Caspase-3酶原逐渐被水解剪切并促进NF-κB活化表达;通过转染SODD小分子干扰质粒的Jurkat白血病细胞对紫杉醇的凋亡敏感性显著高于对照组,差异有统计学意义(F=10.35,P＜0.05).结论紫杉醇可有效诱导白血病细胞凋亡,SODD/Bcl-2基因调控代表了一个独特的紫杉醇类诱导的细胞凋亡信号传导通路,这条通路最终通过激活Caspase-3执行,而NF-κB在这条通路中对相关凋亡基因起反应性调节作用.小分子干扰技术抑制白血病细胞SODD表达后能显著增加紫杉醇的化疗敏感性.
목적 탐토자삼순유도백혈병세포조망적신호도경급사망결구역침묵자(SODD)분자파향대자삼순화료적증민작용.방법 채용Western blot법검측자삼순대인급성림파세포백혈병Jurkat세포SODD、B세포림파류/백혈병-2기인(Bcl-2)、함반광안산적천동안산단백수해매(Caspase-3)、핵전록인자(NF-κB)-P65단백표체적영향;구건1개재체편마3조질립표체재체적SODD소분자간우질립,전염지Jurkat세포,채용전화병정(PI)표기류식세포술검측자삼순대세포적조망유도솔.결과 자삼순재유도Jurkat백혈병세포조망과정중능현저하조SODD급Bcl-2단백적표체,재해조망과정중Caspase-3매원축점피수해전절병촉진NF-κB활화표체;통과전염SODD소분자간우질립적Jurkat백혈병세포대자삼순적조망민감성현저고우대조조,차이유통계학의의(F=10.35,P＜0.05).결론 자삼순가유효유도백혈병세포조망,SODD/Bcl-2기인조공대표료일개독특적자삼순류유도적세포조망신호전도통로,저조통로최종통과격활Caspase-3집행,이NF-κB재저조통로중대상관조망기인기반응성조절작용.소분자간우기술억제백혈병세포SODD표체후능현저증가자삼순적화료민감성.
Objective To explore the signaling pathway of apoptosis induced by Paclitaxel (PTX) in leukemia cells and the chemosensitizing effect of adding short hairpin RNA(shRNA) on PTX,which targets the silencer of death domains(SODD).Methods After being treated with PTX,the expressions of SODD,B-cell lymphoma/leukemia-2 (Bcl-2),nuclear factor kappa B (NF-κB) and Caspase-3 proteins in Jurkat cells were determined by Western blot ;the shRNA-SODD vectors were constructed and transfected into Jurkat cells by electroporation,and then G418 was used to select the stable tranfected cell line expressing the shRNA-SODD recombinant plasmids.The incidence of cell apoptosis induced by PTX was determined by flow cytometry labeled with propidium iodide.Results During the process of inducing apoptosis of Jurkat cells,PTX could significantly down-regulate the expressions of SODD and Bcl-2 proteins,degrade Caspase-3 and activate NF-κB.The apoptotic sensibility of Jurkat cells transfected with shRNA-SODD to PTX was significantly increased compared with the control group,and the difference was statistically significant (F =10.35,P ＜ 0.05).Conclusions PTX can effectively induce apoptosis of Jurkat cells.Perhaps,SODD/Bcl-2 represents a specific apoptotic signaling pathway of PTX in leukemia cells and this apoptotic signaling pathway is Caspase-3-dependent,in which the function of NF-κB is to modulate the correlative apoptotic factors.Inhibiting the expression of SODD through transfecting shRNA-SODD vectors can significantly increase the apoptotic sensibility of leukemia cells to PTX.