Yao K, Xing H C, Wu B, Li Y, Liao A J, Yang W, Liu Z G
Hematology Department of Shengjing Hospital of China Medical University, Shenyang, China.
Anesthesiology Department of Central Hospital Affiliated to Shenyang Medical College, Shenyang, China.
Genet Mol Res. 2015 Mar 20;14(1):1968-74. doi: 10.4238/2015.March.20.6.
We examined the effect of transforming growth factor-b inducible early gene-1 (TIEG1) on the apoptosis of leukemic cell lines and expression of B-cell lymphoma 2 (Bcl-2) and phosphatase and tensin homolog (Pten). Four leukemic cell lines (HL-60, U937, Raji, and K562) were treated with 0, 1, 5, 10, and 20 ng/mL TIEG1, respectively. The cell growth inhibitory ratio was assessed using the MTT assay. An inhibitory curve was drawn, and half-maximal inhibitory concentration was calculated. Additionally, 1640 culture medium containing 10 ng/mL TIEG1 was used to culture leukemic cell lines for 0, 6, 12, 24, and 48 h. The apoptosis of each cell line at different time points was detected by flow cytometry. Total RNA was extracted before reverse transcription-polymerase chain reaction. The products of this reaction were analyzed by electrophoresis, and the expression of Bcl-2/Bcl-2-associated X protein (Bax) and Pten were detected. After treatment with TIEG1, proliferation of the 4 leukemic cell lines was inhibited both time- and dose-dependently. During apoptosis induction, the expression of Bcl-2 was decreased and the expressions of Bax and Pten were increased in the 4 leukemic cell lines induced by TIEG1 (P < 0.05). TIEG1 can inhibit the proliferation of leukemic cells and induce their apoptosis in a time- and dose-dependent manner. A close relationship exists between Bcl-2/Bax and Pten expression and cell apoptosis induced by TIEG1.
我们研究了转化生长因子-β诱导早期基因-1(TIEG1)对白血病细胞系凋亡以及B细胞淋巴瘤2(Bcl-2)和磷酸酶及张力蛋白同源物(Pten)表达的影响。分别用0、1、5、10和20 ng/mL的TIEG1处理四种白血病细胞系(HL-60、U937、Raji和K562)。使用MTT法评估细胞生长抑制率。绘制抑制曲线并计算半数最大抑制浓度。此外,用含有10 ng/mL TIEG1的1640培养基培养白血病细胞系0、6、12、24和48小时。通过流式细胞术检测不同时间点各细胞系的凋亡情况。在逆转录-聚合酶链反应之前提取总RNA。对该反应产物进行电泳分析,检测Bcl-2/Bcl-2相关X蛋白(Bax)和Pten的表达。用TIEG1处理后,四种白血病细胞系的增殖受到时间和剂量依赖性抑制。在诱导凋亡过程中,TIEG1诱导的四种白血病细胞系中Bcl-2表达降低,Bax和Pten表达增加(P<0.05)。TIEG1可抑制白血病细胞的增殖并以时间和剂量依赖性方式诱导其凋亡。Bcl-2/Bax和Pten表达与TIEG1诱导的细胞凋亡之间存在密切关系。