Jiang Tian-Hua, Huang Zhi-Gang
Department of Blood Transfusion, Deyang Municipal People's Hospital of Sichuan Province, Deyang 618000, Sichuan Province, China.
Department of Laboratory Medicine in West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China. E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2017 Feb;25(1):104-109. doi: 10.7534/j.issn.1009-2137.2017.01.018.
To investigate the effects of KCa3.1 channel inhibitor TRAM-34 on the proliferation and invasion of leukemia cell line HL-60.
HL-60 cells at logarithmic growth phase exposed to TRAM-34 at the final concentration of 25, 50, 75 and 100 nmol/L were used as experimental group. The HL-60 cells of control group was cultured in 10% fetal bovine serum-RPMI 1640. The proliferation inhibition rate of TRAM-34 on HL-60 cells was detected by adding MTT solution after 24, 48 and 72 h culture. The cell apoptotic rate and cell cycle distribution of HL-60 cells treated with TRAM-34 were evaluated by flow cytometry with Annexin V-FITC/propidium iodide(PI) double staining or PI single staining. The number of transmembrane cells was detected by Transwell at 24 and 48 h after treatment with TRAM-34. The effect of TRAM-34 on CDK6, P53 and MMP-2 mRNA level was detected by real-time quantitative PCR.
Compared with the control group (0 nmol/L), the inhibition rate, apoptosis rate, G/G phase cell proportion and P53 mRNA level all increased, but the percentages of cells in S phase, cell number penetrating the membrane and mRNA levels of CDK6 and MMP-2 in the TRAM-34-treated group decreased (P<0.05) except for 24 h proliferation rate of TRAM-34 at low concentration (25 nmol/L). The effect of TRAM-34 on the above indices was enhanced with the increase of concentration and prolongation of time, and the differences were statistically significant (P<0.05).
TRAM-34 can inhibit the proliferation and invasion of HL-60 cells, and can induce cell apoptosis and G/G arrest. The time and concentration of TRAM-34 have effect on the malignant behavior of HL-60 cells.
探讨钾通道Ca3.1（KCa3.1）通道抑制剂TRAM - 34对白血病细胞株HL - 60增殖和侵袭的影响。
将对数生长期的HL - 60细胞分别用终浓度为25、50、75和100 nmol/L的TRAM - 34处理作为实验组。对照组HL - 60细胞在含10%胎牛血清的RPMI 1640培养基中培养。培养24、48和72 h后加入MTT溶液检测TRAM - 34对HL - 60细胞的增殖抑制率。采用Annexin V - FITC/碘化丙啶（PI）双染或PI单染的流式细胞术评估TRAM - 34处理后HL - 60细胞的凋亡率和细胞周期分布。TRAM - 34处理24和48 h后，用Transwell检测穿膜细胞数。通过实时定量PCR检测TRAM - 34对细胞周期蛋白依赖性激酶6（CDK6）、P53和基质金属蛋白酶 - 2（MMP - 2）mRNA水平的影响。
与对照组（0 nmol/L）相比，除低浓度（25 nmol/L）TRAM - 34处理24 h的增殖率外，TRAM - 34处理组的抑制率、凋亡率、G0/G1期细胞比例和P53 mRNA水平均升高，而S期细胞百分比、穿膜细胞数以及CDK6和MMP - 2的mRNA水平降低（P<0.05）。TRAM - 34对上述指标的影响随浓度增加和时间延长而增强，差异有统计学意义（P<0.05）。
TRAM - 34可抑制HL - 60细胞的增殖和侵袭，并可诱导细胞凋亡和G0/G1期阻滞。TRAM - 34的作用时间和浓度对HL - 60细胞的恶性行为有影响。
