摘要
目的探讨阻断信号转导与转录激活因子3(STAT3)信号通路对恶性淋巴瘤细胞凋亡的影响。方法用0、200、400、800、1600 nmol/L的STAT3信号通路特异性阻断剂JSI-124作用于恶性淋巴瘤细胞株Raji,噻唑蓝(MTT)法检测细胞增殖水平,计算半数抑制浓度。用半数抑制浓度的JSI-124作用于Raji细胞,流式细胞术检测细胞凋亡和细胞周期,Western blot法检测细胞周期蛋白D1(cyclin D1)、STAT3、磷酸化的STAT3(p-STAT3)、活化的半胱氨酸天冬氨酸蛋白水解酶3(cleaved caspase 3)的蛋白相对表达水平。结果随着JSI-124作用浓度的升高,细胞存活率逐渐下降。计算半数抑制浓度为(615.62±52.98)nmol/L,故后续选用600 nm/L的JSI-124作用于恶性淋巴瘤细胞。600 nm/L作用组细胞凋亡率明显高于0 nm/L作用组(P﹤0.01)。600 nm/L作用组G0/G1期细胞所占比例明显低于0 nm/L作用组,G2/M期细胞所占比例明显高于0 nm/L作用组,差异均有统计学意义(P﹤0.01);两组S期细胞所占比例比较,差异无统计学意义(P﹥0.05)。600 nm/L作用组p-STAT3、cyclin D1蛋白相对表达水平明显低于0 nm/L作用组,cleaved caspase 3蛋白相对表达水平明显高于0 nm/L作用组,差异均有统计学意义(P﹤0.01)。结论阻断STAT3信号通路可促进恶性淋巴瘤细胞凋亡,抑制细胞增殖,将细胞周期阻滞在G2/M期。
Objective To investigate the effect of signal transducer and activator of transcription 3(STAT3)signaling pathway on the apoptosis of malignant lymphoma cells.Method Raji,a malignant lymphoma cell line,was treated with JSI-124,a specific blocker of STAT3 signaling pathway,at the concentration of 0,200,400,800 and 1600 nmol/L.MTT assay was used to detect cell proliferation.The half inhibitory concentration was calculated.Raji cells were treated with JSI-124 at half inhibitory concentration.Cell apoptosis and cell cycle were detected by flow cytometry,besides,the relative expression levels of cyclin D1,STAT3,phosphorylated STAT3(p-STAT3)and cleaved caspase 3 were determined by Western blot.Result With the increase of JSI-124 concentration,cell viability declined gradually,the half inhibitory concentration was calculated as(615.62±52.98)nmol/L,which therefore was used to treat the malignant lymphoma cells.The apoptosis rate in 600 nm/L group was significantly higher than that in 0 nm/L group(P<0.01).The percentage of G0/G1 phase cells in 600 nm/L group was significantly lower than that in 0 nm/L group,while the proportion of G2/M phase cells was significantly higher in 600 nm/L group than that of 0 nm/L group(P<0.01),there was no significant difference in regard to the proportion of S phase cells(P >0.05).The relative expression levels of p-STAT3 and cyclin D1 protein in 600 nm/L group were significantly lower than those in 0 nm/L group(P<0.01),and the cleaved caspase 3 protein was significantly higher than that of 0 nm/L group,the difference was statistically significant(P<0.01).Conclusion Blocking STAT3 signaling pathway can promote the apoptosis of malignant lymphoma cells,inhibit cell proliferation and block cell cycle in G2/M phase.
作者
姚金晓
杨如玉
马海龙
李超
魏旭东
YAO Jinxiao;YANG Ruyu;MAHailong;LI Chao;WEI Xudong(Department of Hematology,Nanyang Central Hospital,Zhengzhou 473000,He’nan,China;Department of Hematology,Cancer Hospital Affiliated to Zhengzhou University,Zhengzhou 450008,He’nan,China)
出处
《癌症进展》
2019年第2期227-230,共4页
Oncology Progress
