摘要
目的探讨AMD3100、利妥昔单抗(Rituximab)对人弥漫大B细胞淋巴瘤细胞株SU-DHL-2增殖、凋亡与侵袭能力的影响及其分子机制。方法对弥漫大B细胞淋巴瘤SU-DHL-2细胞株进行体外培养,将对数生长期细胞分为对照组(未经任何处理)、AMD3100组(加入1μmol/L AMD3100)、Rituximab组(加入10μg/mLRituximab)、联合用药组(加入1μmol/LAMD3100,孵育1 h后加入10μg/mLRituximab)。采用细胞计数(Cellcountingkit-8,CCK-8)增殖实验检测细胞增殖、流式细胞术检测细胞凋亡、Transwell小室检测细胞迁移和侵袭、Westernblot法检测SU-DHL-2细胞株中趋化因子受体4(CXCR4)、蛋白激酶B(AKT)、磷酸化蛋白激酶B(pAKT)、基质金属蛋白酶9(MMP-9)、基质金属蛋白酶2(MMP-2)蛋白质表达情况及外源性AMD3100、Rituximab及联合用药后蛋白质表达水平的变化。结果细胞增殖实验结果显示,AMD3100组、Rituximab组均可显著抑制SU-DHL-2细胞株的增殖,差异具有统计学意义(P<0.05)。联合用药组具有协同作用,使用后细胞存活率(59.431%)显著降低,差异具有统计学意义(P<0.01)。细胞凋亡实验结果显示,AMD1300组、Rituximab组与对照组比较,联合用药组与AMD3100组、Rituximab组与对照组比较,细胞凋亡率显著升高,差异具有统计学意义(P<0.01)。Transwell小室实验结果显示,对照组穿膜相对细胞数(100.000±0.526、100.000±0.432)高于AMD3100组(85.253±0.530、86.943±0.592)、Rituximab组(79.743±0.531、81.073±0.433)、联合用药组(61.860±0.526、63.477±0.592),各组间比较差异具有统计学意义(P均<0.01)。Westernblot结果显示,AMD3100组、Rituximab组CXCR4、AKT、p-AKT、MMP-9、MMP-2表达水平降低,联合用药后作用增强,差异具有统计学意义(P<0.01)。结论AMD3100与Rituximab可能通过调节MMP-9、MMP-2及AKT信号通路,对SU-DHL-2细胞株的增殖、促进、迁移和侵袭起调节作用,并具有协同效应。
Objective To investigate the effects of AMD3100 and Rituximab on proliferation,apoptosis and invasion of human diffuse large B-cell lymphoma cell line SU-DHL-2 and it molecular mechanisms.Methods Diffuse large Bcell lymphoma cell line SU-DHL-2 cells were cultured in vitro,and logarithmic growth phase cell lines and were di⁃vided into the control group(without any treatment),AMD3100 group(adding 1μmol/L AMD3100),Rituximab group(Add 10μg/mL Rituximab),and two-drug combination group(add 1μmol/L AMD3100 to the cells,add 10μg/mL Rituximab after 1 h incubation).Using CCK-8 to detected cell proliferation,flow cytometry to detected cell apoptosis,Transwell chamber to detected migration and invasion,and Western blot to detected CXCR4,AKT,pAKT,MMP-9,MMP-2 protein expression in SU-DHL-2 cells,and foreign changes in protein expression levels of AMD3100 and Rituximab after single-drug and combined use。Results In cell proliferation experiments,AMD3100 and Rituximab can significantly inhibit the proliferation of SU-DHL-2 cells,the difference was statistically significant(P<0.05).The combination of the two drugs had a synergistic effect and cell survival rate(59.431%)was significantly reduced after the combined use,and the difference was statistically significant(P<0.01).In the apoptosis experiment,compared with the single-drug group and the control group,the combined drug group was compared with the single-drug group and the control group,the apoptosis rate was significantly higher,and the difference was statistically significant(P<0.01).In the Transwell chamber migration and invasion experiment,the relative cell numbers(100.000±0.526,100.000±0.432)in the control group were higher than those in the AMD3100 group(85.253±0.530;86.943±0.592)and Ritux⁃imab group(79.743±0.531;81.073±0.433),the two-drug combination group(61.860±0.526;63.477±0.592),the sin⁃gle-drug group was compared with the control group,the combination group was compared with the single-drug group and the control group,the difference was statistically significant(both P<0.01).Western blot,the expression levels of CXCR4,AKT,pAKT,MMP-9 and MMP-2 in the AMD3100 and Rituximab single-drug treatment group was de⁃creased,and the combined application of the two drugs was increased in effect.,the difference was statistically signifi⁃cant(P<0.01).Conclusion AMD3100 and Rituximab may regulate the proliferation,promotion,migration and invasion of SU-DHL-2 cells by regulating MMP-9,MMP-2 and AKT signaling pathways,and it have synergistic effects.
作者
马志萍
曹燕珍
李新霞
MA Zhiping;CAO Yanzhen;LI Xinxia(Department of Pathology,the First Affiliated Hospital,Urumqi 830054,China;Department of Pathology,Affiliated Tumor Hospital of Xinjiang Medical University,Urumqi 830011,China)
出处
《新疆医科大学学报》
CAS
2022年第4期359-364,共6页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区卫生计生委青年医学科技人才专项(WJWY-201904)。
