MRP8/MRP14对宫颈癌细胞增殖的影响及机制

Effects of MRP8/MRP14 on proliferation of cervical cancer cells and the mechanism

目的探讨髓样相关蛋白8/14异源二聚体(MRP8/MRP14)对宫颈癌细胞增殖的影响及机制。方法体外培养人宫颈癌Hela细胞,加入MRP8/MRP14(观察组),另设对照组加入等体积PBS,于培养12、24、48、72 h分别采用CCK8法检测两组细胞增殖能力。取Hela细胞进行核质分离,加入MRP8/MRP14共培养30 min,采用Western blot法检测细胞凋亡相关信号通路蛋白p38 MAPK、JNK及细胞增殖相关信号通路蛋白ERK1/2、NF-κB表达。将Hela细胞分为MRP8/MRP14组和p38 MAPK、JNK、ERK1/2、NF-κB抑制剂组,RP8/MRP14组加入MRP8/MRP14;抑制剂组先分别加入p38 MAPK、JNK、ERK1/2、NF-κB抑制剂预处理2 h,再加入MRP8/MRP14;采用CCK8法观察各组细胞增殖能力。结果观察组加入MRP8/MRP14培养12、24、48、72 h后,细胞增殖能力均高于对照组,且随时间延长,细胞增殖能力逐渐增强(P均<0.05)。观察组加入MRP8/MRP14培养30 min后,与对照组比较,p38 MAPK、JNK、ERK1/2活性明显升高,NF-κB在细胞核中的表达升高(P均<0.05)。p38 MAPK、JNK抑制剂组细胞增殖能力较MRP8/MRP14组增强;ERK、NF-κB抑制剂组细胞增殖能力较MRP8/MRP14组减弱(P均<0.05)。结论 MRP8/MRP14能够促进Hela细胞增殖,其机制与p38 MAPK、JNK、ERK1/2及NF-κB等多条相关信号通路被激活有关。

Objective To investigate the effect of myeloid-related protein-8 /14 heterodimer（ MRP8 / MRP14） on the proliferation of cervical cancer Hela cells and the possible mechanism. Methods Cervical cancer cell line Hela was cultured in vitro and treated with MRP8 / MRP14（ observation group） or PBS（ control group）,then we measured the proliferation ability at 12,24,48 and 72 h with CCK8. The nuclear of Hela cells was separated and then Hela cells were cultured with MRP8 / MRP14 for 30 min. The expression of apoptosis-related signaling pathway protein p38 MAPK,JNK and proliferation-related signaling pathway protein ERK1 /2 and NF-κB was measured by Western blotting. Hela cells were divided into MRP8 / MRP14 group,p38 MAPK,JNK,ERK1 /2 and NF-κB inhibitor group. Hela cells in the MRP8 / MRP14 group were only treated with MRP8 / MRP14,and the inhibitor groups were pretreated with p38 MAPK,JNK,ERK1 /2 or NF-κB inhibitor for 2 h then were incubated with MRP8 / MRP14. The proliferation ability of Hela cells were detected by CCK8.Results Compared with the control group,MRP8 / MRP14 enhanced the Hela cell proliferation ability at 12,24,48 and72 h,and the proliferation ability increased gradually as time passed in the observation group（ all P 0. 05）. After the treatment of MRP8 / MRP14 for 30 min,p38 MAPK,JNK and ERK1 /2 were activated and the expression of NF-κB in the nucleus was increased（ all P 0. 05）. In the p38 MAPK and JNK inhibitor groups,the proliferation ability of Hela cells were increased,while the proliferation ability of Hela cells were decreased in the ERK and NF-κB inhibitor groups as compared with that of the MRP8 / MRP14 group（ all P 0. 05）. Conclusion MRP8 / MRP14 can promote the proliferation of Hela cells and this mechanism may be related with the activation of p38,JNK,ERK1 /2 and NF-κB signaling pathways.