临床肿瘤学杂志

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Rho激酶抑制剂Y27632对非小细胞肺癌增殖、凋亡及侵袭转移的影响

朱颖1,刘渤娜2   

  1. 1 110042沈阳辽宁省肿瘤医院药剂科2 110812解放军第202医院肿瘤介入科
  • 收稿日期:2015-12-13 修回日期:2016-01-20 出版日期:2016-03-30 发布日期:2016-03-30
  • 通讯作者: 刘渤娜

Effects of Rho kinase inhibitor Y27632 on proliferation, apoptosis and invasion of non-small cell lung cancer

ZHU Ying,LIU Bona   

  1. Department of Pharmacy,Tumor Hospital of Liaoning Province,Shenyang 110042,China
  • Received:2015-12-13 Revised:2016-01-20 Online:2016-03-30 Published:2016-03-30
  • Contact: LIU Bona

摘要: 目的 探讨Rho激酶抑制剂Y27632对非小细胞肺癌细胞增殖、凋亡及侵袭转移的影响。方法 采用5、10、20、40、80 μmol/L Y27632处理非小细胞肺癌细胞A549后,设不加药物的为对照组。分别在24、48、72 h 3个不同刺激时间点应用四甲基偶氮唑盐比色法(MTT法)检测A549细胞的吸光度并计算增殖抑制率和半数抑制浓度(IC50),膜联蛋白-增强型绿色荧光蛋白(Annexin-EGFP)/碘化丙啶(PI)双染流式细胞术检测Y27632处理24、48 h后的细胞凋亡情况,PI单染流式细胞术检测Y27632处理48 h后细胞周期各时相(G0/G1、S和G2/M期)分布情况,Transwell试验检测Y27632处理48 h后的穿膜细胞数。结果 Y27632 可抑制A549的增殖(与对照组比较,P<0.05),呈剂量和时间依赖性,且IC50值随作用时间的延长而降低;与对照组比较,Y27632处理后的凋亡率、G0/G1期细胞比例均升高,而S、G2/M期细胞比例均降低,各浓度间的差异均有统计学意义(P<0.05);Transwell实验发现Y27632可抑制A549细胞的侵袭,如0、5、10、20、40、80 μmol/L Y27632处理后的穿膜细胞数依次为(78.1±6.0)、(62.3±5.7)、(51.7±5.2)、(42.3±6.9)、(36.4±5.3)和(22.8±4.6)个,差异均有统计学意义(P<0.05)。结论Y27632可抑制A549细胞的增殖并诱导凋亡及G0/G1期阻滞,同时可抑制细胞侵袭,在肺癌治疗上有一定价值。

Abstract:

Objective To investigate the effect of Rho kinase inhibitor Y27632 on cell proliferation,apoptosis and invasion and metastasis of non-small cell

lung cancer. Methods The non-small cell lung cancer cells A549 were treated with different concentrations of Y27632(5, 10, 20, 40, 80 μmol/L) and the

cells without treatment were set as the control group. The absorbance of A549 cells was detected by MTT method at 24,48,72 h after stimulation, and the

inhibitory rates of proliferation and half inhibitory concentrations(IC50)were calculated accordingly. The cell apoptotic rates were measured at 24 and 48 h by Annexin-enhanced green fluorescent protein (Annexin-EGFP)/iodinated propidium (PI) double staining via flow cytometry. The distribution of cell cycle, including G0/G1, S and G2/M, were detected by PI single staining with flow cytometry. The number of cells penetrating the membrane was detected by Transwell test. Results Y27632 can inhibit the proliferation of A549 (compared with the control group P<0.05) in a dose- and time-dependence manner, and the IC50 value decreased with the prolongation of the time. Compared with the control group, the apoptotic rates and G0/G1 phase cell proportions were increased, and the proportions of S and G2/M cells were decreased, and the differences among different concentrations were statistically significant (P<0.05). Transwell experiments showed that Y27632 could inhibit the invasion of A549 cells. For instance, the number of cells penetrating the membrane were (78.1±6),(62.3±5.7),(51.7±5.2),(42.3±6.9),(36.4±5.3) and(22.8±4.6) for treatment with 0, 5, 10, 20, 40,80 mol/L Y27632, respectively. Conclusion In addition to the inhibition effect on cell invasion, Y27632 can inhibit the proliferation of A549 cells and induce apoptosis and G0/G1 arrest,having a certain value in the treatment of lung cancer.

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