CAJ | 학술논문

目的研究黄芪多糖(APS)对人红白血病K562细胞增殖的抑制作用及机制.方法用不同质量浓度APS(0 mg/L、100 mg/L、200 mg/L和400 mg/L)处理K562细胞(购于中科院上海细胞库)后,通过四甲基偶氮唑盐(MTT)法观察APS对K562细胞生长速度和倍增时间的影响,通过流式细胞术检测APS对K562细胞周期分布的影响,通过反转录-聚合酶链反应(RT-PCR)和Western blotting方法检测细胞周期蛋白A(Cyclin A)、细胞周期蛋白B(Cyclin B)、细胞周期蛋白E(Cyclin E)和p21基因在mRNA和蛋白水平的表达.结果 MTT结果显示:与对照组相比,100 mg/L、200 mg/L和400 mg/L APS处理的K562细胞生长速度均明显减慢(P均＜0.01),倍增时间显著延长(P均＜0.01).流式细胞术检测周期分布结果显示:与对照组相比,APS组K562细胞G1期细胞显著增多,差异有统计学意义(P＜0.01),而S期和G2/M期细胞显著减少,差异有统计学意义(P＜0.01).RT-PCR和Western blotting结果显示:与对照组相比,APS组K562细胞中Cychn B和Cyclin E 在mRNA和蛋白的表达均显著降低,差异均有统计学意义(P均＜0.01);p21在mRNA和蛋白的表达均显著增加,差异均有统计学意义(P均＜0.01);而Cyclin A在mRNA和蛋白的表达差异均无统计学意义(P均＞0.05).结论 APS能够显著抑制人红白血病K562细胞的增殖能力,APS可能是通过下调Cyclin B和Cyclin E的表达及上调p21的表达而抑制K562细胞的增殖能力.
목적 연구황기다당(APS)대인홍백혈병K562세포증식적억제작용급궤제.방법 용불동질량농도APS(0 mg/L、100 mg/L、200 mg/L화400 mg/L)처리K562세포(구우중과원상해세포고)후,통과사갑기우담서염(MTT)법관찰APS대K562세포생장속도화배증시간적영향,통과류식세포술검측APS대K562세포주기분포적영향,통과반전록-취합매련반응(RT-PCR)화Western blotting방법검측세포주기단백A(Cyclin A)、세포주기단백B(Cyclin B)、세포주기단백E(Cyclin E)화p21기인재mRNA화단백수평적표체.결과 MTT결과현시:여대조조상비,100 mg/L、200 mg/L화400 mg/L APS처리적K562세포생장속도균명현감만(P균＜0.01),배증시간현저연장(P균＜0.01).류식세포술검측주기분포결과현시:여대조조상비,APS조K562세포G1기세포현저증다,차이유통계학의의(P＜0.01),이S기화G2/M기세포현저감소,차이유통계학의의(P＜0.01).RT-PCR화Western blotting결과현시:여대조조상비,APS조K562세포중Cychn B화Cyclin E 재mRNA화단백적표체균현저강저,차이균유통계학의의(P균＜0.01);p21재mRNA화단백적표체균현저증가,차이균유통계학의의(P균＜0.01);이Cyclin A재mRNA화단백적표체차이균무통계학의의(P균＞0.05).결론 APS능구현저억제인홍백혈병K562세포적증식능력,APS가능시통과하조Cyclin B화Cyclin E적표체급상조p21적표체이억제K562세포적증식능력.
Objective To explore the inhibitory effect of astragalus polysaccharide (APS) on the proliferation of human erythroleukemia K562 cells and its mechanisms.Methods After K562 cells (purchased from Shanghai cell bank of chinese academy of science) were treated with different concentrations (0 mg/L,100 mg/L,200 mg/L and 400 mg/L) of APS.The influences of APS on the growth rate,doubling time and cell cycle distribution of K562 cells were observed by methyl thiazolyl tetra-zolium assay (MTF) and flow cytometry,respectively.Furthermore,the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting assay were used to detect the expressions of Cyclin A,Cyclin B,Cyclin E and p21 gene at the mRNA and protein levels,respectively.Results MTT assay findings showed that,compared to the control group (0 mg/L APS),growth rates of K562 cells treated with 100 mg/L,200 mg/L and 400 mg/L APS decreased significantly (all P ＜ 0.01),and the doubling times lengthened significantly (all P ＜ 0.01).Flow cytometry findings revealed that,compared to the control group,the G1 phase cells in K562 cells of APS group increased significantly (P ＜0.01),while the S and G2/M phase cells decreased significantly (all P ＜ 0.01).RT-PCR and Western blotting results indicated that Cyclin B and Cyclin E expression of K562 cells at the mRNA and protein levels in the APS group were significantly lower than those of the control group(all P ＜ 0.01),whereas p21 expression was significantly enhanced at mRNA and protein levels (P ＜ 0.01),and Cyclin A expression was not significantly different at mRNA and protein levels between the 2 groups (all P ＞ 0.05).Conclusions APS could inhibit the proliferation of human erythroleukemia K562 cells.APS could inhibit the proliferation of K562 cells by down-regulating the expression of Cyclin B and Cyclin E and up-regulating the expression of p21.