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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(P<0.01);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(P<0.01),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。
OBJECTIVE: To investigate the effect of total flavonoids of Phragmites australis (FGVL) against the acute promyelocytic leukemia NB4 cell line, and to provide a new basis for clinical treatment of promyelocytic leukemia. Methods: The acute promyelocytic leukemia cell line NB4 was divided into three experimental groups (FGVL concentration of 50,100,200 mg · L -1) and blank group (same volume of culture medium) The effect of FGVL on the growth of NB4 cells was observed by trypan blue staining. The inhibitory effect of FGVL on cell proliferation was detected by MTT assay. The apoptosis of NB4 cells was observed by acridine orange / ethidium bromide (EB / EB) staining The apoptosis of Bcl-2 and Bcl-2 were detected by RT-PCR, and the changes of apoptosis were analyzed by DNA agarose gel electrophoresis. X protein (Bax) mRNA expression. Results: Compared with the blank group, FGVL (50, 100, 200 mg · L-1) inhibited the proliferation of NB4 cells (P <0.01). The morphological features of 24 h showed obvious apoptosis and DNA agarose gel electrophoresis showed typical The results of RT-PCR indicated that FGVL downregulated the expression of Bcl-2 mRNA and up-regulated the expression of Bax mRNA (P < 0.01), its role and dose was positively correlated. CONCLUSION: FGVL can inhibit the proliferation of NB4 cells and induce the apoptosis of NB4 cells. Its mechanism is related to the regulation of Bcl-2 / Bax expression, which may provide a new possibility for the clinical chemotherapy of acute promyelocytic leukemia.