TRAM-34对白血病细胞株HL-60细胞增殖及侵袭的影响

发布时间：2018-06-19 03:12

本文选题：KCa.通道抑制剂 + TRAM-　；参考：《中国实验血液学杂志》2017年01期

【摘要】：目的:探讨KCa3.1通道抑制剂TRAM-34对白血病细胞株HL-60细胞增殖及侵袭的影响。方法:取对数生长期HL-60细胞,给予实验组分别加入终浓度25、50、75、100 nmol/L TRAM-34的培养液,给予对照组加入含10%胎牛血清RPM11640培养液,同步培养24、48和72 h后加入MTT溶液以检测TRAM-34对HL-60细胞的增殖抑制率;在TRAM-34处理后的每个浓度收集细胞1×10~6个,采用膜联蛋白V-异硫氰酸荧光素(Annexin VFITC)/碘化丙啶(PI)双染或PI单染流式细胞术分别检测TRAM-34处理后的细胞凋亡率和细胞周期时相分布情况,采用Transwell检测TRAM-34各浓度24、48 h处理后的穿膜细胞数,采用实时定量PCR检测TRAM-34对CDK6、P53及MMP-2 mRNA水平的影响。结果:与对照组相比(0 nmol/L),除低浓度(25 nmol/L)TRAM-34处理24 h对HL-60细胞增殖无影响外,其余浓度和作用时间的增殖抑制率、凋亡率、G_0/G_1期细胞比例及p53 mRNA水平均升高,S期细胞比例、穿膜细胞数及CDK6、MMP-2 mRNA水平均降低,而且以上差异均有统计学意义(P0.05)。TRAM-34对HL-60以上指标的作用效果随浓度的升高和作用时间的延长而增强,各浓度间的差异具有统计学意义(P0.05)。结论:TRAM-34可抑制HL-60细胞的增殖及侵袭能力,同时可诱导细胞凋亡及G0/G1期阻滞,且TRAM-34作用的时间和浓度均对HL-60细胞恶性行为有影响。
[Abstract]:Objective: To investigate the effect of KCa3.1 channel inhibitor TRAM-34 on the proliferation and invasion of leukemia cell line HL-60. Methods: the logarithmic growth period HL-60 cells were taken, and the experimental group was added to the final concentration of 25,50,75100 nmol/L TRAM-34, and the control group was added with the 10% fetal bovine serum RPM11640 culture solution, and the synchronous culture of 24,48 and 72 h was added. The proliferation inhibition rate of TRAM-34 to HL-60 cells was detected by MTT solution; 1 x 10~6 cells were collected at each concentration after TRAM-34 treatment. The apoptosis rate and phase distribution of cell cycle were detected by annexin V- isothiocyanate (Annexin VFITC) / PI (PI) double dye or PI single dye flow cytometry. The effects of TRAM-34 on the level of CDK6, P53 and MMP-2 mRNA were detected by real-time quantitative PCR, and the effects of TRAM-34 on the level of CDK6, P53 and MMP-2 mRNA were detected by Transwell. Results: compared with the control group (0 nmol/L), the proliferation and inhibition of the remaining concentration and time were compared with the control group (0 nmol/L) except the low concentration (25 nmol/L). The rate of production, the rate of apoptosis, the proportion of G_0/G_1 cells and the level of p53 mRNA were all increased, the proportion of S cells, the number of membrane cells and the level of CDK6 and MMP-2 mRNA were all decreased, and the above differences were statistically significant (P0.05) the effect of.TRAM-34 on the index of HL-60 increased with the increase of concentration and the prolongation of action time. Statistical significance (P0.05). Conclusion: TRAM-34 can inhibit the proliferation and invasion of HL-60 cells, and induce apoptosis and G0/G1 phase arrest, and the time and concentration of TRAM-34 effect on the malignant behavior of HL-60 cells.
【作者单位】：四川省德阳市人民医院输血科;四川大学华西医院实验医学科;
【分类号】：R733.7

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