摘要
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（0mg／L、100mg／L、200mg／L和400mg／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（CyclinA）、细胞周期蛋白B（CyclinB）、细胞周期蛋白E（CyclinE）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，100mg／L、200mg／L和400mg／LAPS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Cyclin 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可能是通过下捌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。
Objective To explore the inhibitory effect of astragalus polysaccharide （APS） on the proliferation of human erythroleukemia K562 ceils 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-znlium assay （MTT）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 I（562 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 ceils. APS could inhibit the proliferation of K562 ceils by down-regulating the expression of Cyclin B and Cyclin E and up-regulating the expression of p21.
出处
《中华实用儿科临床杂志》
CAS
CSCD
北大核心
2014年第12期936-939,共4页
Chinese Journal of Applied Clinical Pediatrics
