[1]马莎,徐开林,陈翀,等.ADAM10抑制剂GI254023X对CCRF-CEM和Molt4细胞增殖与凋亡的作用及机制研究*[J].徐州医科大学学报,2020,40(07):469-475.
　Effect of ADAM10 inhibitorGI254023X on Proliferation and Apoptosis of Acute T-Lymphoblastic Leukemia CCRF-CEM and Molt4 Cells and Its Possible Mechanisms[J].Journal of Xuzhou Medical University,2020,40(07):469-475.
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ADAM10抑制剂GI254023X对CCRF-CEM和Molt4细胞增殖与凋亡的作用及机制研究*()
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《徐州医科大学学报》[ISSN:2096-3882/CN:32-1875/R]

卷:
40
期数:
2020年07期
页码:
469-475
栏目:
出版日期:
2020-07-30

文章信息/Info

Title:
Effect of ADAM10 inhibitorGI254023X on Proliferation and Apoptosis of Acute T-Lymphoblastic Leukemia CCRF-CEM and Molt4 Cells and Its Possible Mechanisms
作者:
马莎; 徐开林; 陈翀; 王雪; 吴庆运; 曾令宇; 李振宇
文献标志码:
A
摘要:
目的：本研究旨在探讨ADAM10抑制剂GI254023X对急性T淋巴细胞白血病CCRF-CEM及Molt4细胞株增殖、凋亡的影响及其作用机制。方法：选择不同浓度的GI254023X处理CCRF-CEM及Molt4细胞，CCK-8法检测并绘制增殖抑制曲线，Annexin V/7-AAD双标记法分析细胞凋亡情况，流式细胞术检测细胞周期的变化，Western blot检测切割后的Notch1蛋白（Cleaved Notch1），定量PCR检测抑凋亡基因Bcl-2、c-Myc、Mcl-1，促凋亡基因BAD、BAK及Notch1靶基因Hes-1的转录变化。结果：GI254023X能明显抑制CCRF-CEM及Molt4细胞增殖，且抑制效应呈一定的时间和剂量依赖性(CCRF-CEM 细胞24、48 h的相关系数r分别为0.981、0.962, Molt4细胞24、48 h的相关系数 r分别为0.992、0.957)，CCRF-CEM细胞24、48 h的半数抑制浓度（IC50）分别为（29.403±3.28）μmol/L和（11.14±2.21） μmol/L；Molt4细胞24、48 h的半数抑制浓度（IC50）分别为（19.042±1.58）μmol/L和（11.25±2.31）μmol/L；GI254023X能够诱导CCRF-CEM及Molt4细胞凋亡，使细胞阻滞于G0/G1期；Western blot检测结果显示，Cleaved Notch1在GI254023X作用后水平下降；GI254023X能够使抑凋亡基因Bcl-2、c-Myc、Mcl-1和Hes-1表达减低，促凋亡基因BAD、BAK表达升高。结论：GI254023X能抑制CCRF-CEM及Molt4细胞增殖并诱导其凋亡。其机制可能与抑制Notch1活化有关。
Abstract:
ob＜x＞jective: To investigate the effects of ADAM10 inhibitor GI254023X on the proliferation and apoptosis of acute T-lymphoblastic leukemia CCRF-CEM and Molt4 Cells and its mechanisms．Methods: CCRF-CEM and Molt4 Cells were treated with different concentrations of GI254023X. Proliferation-inhibition Acurve was assayed and plotted by CCK-8 method．cell viability and apoptosis was detected by flow cytometry with Annexin V and 7-AAD staining．Cell cycle changes were analyzed by flow cytometry.The cleavage of Notch1 protein was determined by Western blot．The transc＜x＞ripts of anti-apoptotic genes Bcl-2、c-Myc、Mcl-1 ,pro-apoptosis genes BAD、BAK and Notch1 target gene Hes-1 were detected by Real-time PCR ．Results: GI254023X inhibited the proliferation of CCRF-CEM and Molt4 Cells in a time-and dose-depended manner（the r valuesof CCRF-CEM cells for 24 and 48 h were 0.981 and 0.962 respectively, while r values of Molt4 Cells for 24 and 48 h were 0.992 and 0.957 respectively. ）, and the IC50 values of CCRF-CEM cells for 24 and 48 h were （29.403±3.28）μmol/L and （11.14±2.21）μmol/L respectively, while IC 50 values of Molt4 Cells for 24 and 48 h were （19.042±1.58）μmol/L and （11.25±2.31）μmol/L respectively. As compared with the control group ，the apoptosis of cells increased with concentration of GI254023X; Cell cycles were arrested at the G0/G1 phase.Compared with DMSO in control group,The ex＜x＞pression of Cleaved Notch1 was down-regulated after the treatment with GI254023X ．The mRNA levels of anti-apoptotic genes Bcl-2、c-Myc、Mcl-1 and Hes-1 mRNA transc＜x＞ripts in CCRF-CEM and Molt4 Cells were reduced in GI254023X treated group, and the mRNA levels of pro-apoptosis genes BAD、BAK were increased. Conclusion: GI254023X can remarkably inhibit cell proliferation and induce apoptosis in CCRF-CEM and Molt4 Cells．Its mechanism may be associated with inhibition of Notch1 activation
更新日期/Last Update: 2020-07-29