蓬子菜总黄酮诱导急性早幼粒细胞白血病NB4细胞株凋亡的研究

目的:探讨蓬子菜总黄酮(FGVL)抗急性早幼粒细胞白血病NB4细胞株的作用,为以后临床治疗早幼粒细胞白血病提供一个新的依据。方法:以急性早幼粒细胞白血病细胞株NB4为研究对象,实验分为3个实验组(FGVL质量浓度依次为50,100,200 mg·L~(-1))和空白组(相同体积的培养液),通过台盼蓝染色观察FGVL对NB4细胞生长的影响;噻唑蓝法(MTT)检测FGVL对细胞增殖的抑制作用;吖啶橙/溴乙锭(AO/EB)荧光染色观察NB4细胞凋亡的形态学变化;DNA琼脂糖凝胶电泳检测细胞凋亡;逆转录-聚合酶链式反应(RT-PCR)检测凋亡相关基因B细胞淋巴瘤/白血病-2(Bcl-2),Bcl-2相关X蛋白(Bax)mRNA的表达。结果:与空白组比较,FGVL(50,100,200 mg·L-1)能抑制NB4细胞的增殖(P0.01);24 h细胞形态学可见明显的细胞凋亡特征以及DNA琼脂糖凝胶电泳显示典型的DNA"梯形"凋亡条带;RT-PCR实验表明,与空白组比较,FGVL下调了抑制细胞凋亡基因Bcl-2 mRNA的表达,并上调促凋亡基因Bax mRNA的表达(P0.01),其作用与剂量呈正相关。结论:FGVL抗急性早幼粒细胞白血病NB4细胞的增殖并诱导细胞凋亡,其作用机制与调节Bcl-2/Bax表达有关,可为急性早幼粒细胞白血病临床化疗用药提供一个新的可能。

Flavone from Galii Veri Herba et Radix-induced Apoptosis of Acute Promyelocytic Leukemia Cells (NB4)

Objective: To study the effect of flavone from Galii Veri Herba et Radix (FGVL) against acute promyelocytic leukemia cell line NB4, in order to provide a new evidence for future clinical treatment for acute promyelocytic leukemia. Method: In this study, the growth of acute promyelocytic leukemia cell line NB4 at different dosages of FGVL was observed with trypan blue staining. Methyl thiazoly1 terazolium(MTT) assay was used to detect the proliferation inhibition of NB4 cells. Acridine orange ethidium bromide (AO/EB) staining was used to detect morphological changes in NB4 apoptosis. DNA agarose electrophoresis was adopted to test apoptosis. The expression of pro-and anti-apoptotic genes Bax and Bcl-2 were examined by reverse transoription-polymerase chain reaction(RT-PCR). Result: Compared with the blank group, FGVL (50, 100, 200 mg·L^-1) could inhibit the proliferation of NB4 cells(P < 0.01);morphology analysis presented the characteristics of apoptosis, and DNA agarose electrophoresis showed the typical DNA 1adder in 24 h. Accoridng to RT-PCR experiments, compared with the blank group, FGVL could down-regulate the mRNA expression of Bcl-2 and up-regulate the mRNA expression of Bax in NB4 cells (P < 0.01), with the positive correlation with the dosage. Conclusion: FGVL can resist acute promyelocytic leukemia NB4 cell proliferation and induce apoptosis, and its mechanism is related to the expression of Bcl-2/Bax. It provides a new possible clinical chemotherapy for acute promyelocytic leukemia.