IL-6对人卵巢癌细胞体外增殖及锚定非依赖生长能力的影响  

Effects of interleukin-6 on proliferation and anchorage-independent growth of human ovarian cancer cells in vitro

作　　者：毛立群[1,2] 王丹[2] 郭小芹[2] 杨静[2] 屈野[2] 李玲[3] 王越[1,2] 

机构地区：[1]天津市职业与环境危害生物标志物重点实验室,300309 [2]天津武警后勤学院病原生物学与免疫学教研室,300309 [3]天津武警后勤学院药理学教研室,300309

出　　处：《免疫学杂志》2015年第10期835-841,共7页Immunological Journal

基　　金：国家自然科学基金(81041071;81273520;81572852;81502256);天津市自然科学基金重点项目(12JCZDJC26300);武警后勤学院科学技术研究项目(WHZ201202;WHB201404;WHB201405;WHB201406;2015ZXKF05;WHB201505;WHJ201508)

摘　　要：目的研究内源性IL-6对卵巢癌(ovarian cancer,OVCA)细胞体外增殖及锚定非依赖生长能力的影响并探讨其作用机制。方法应用前期建立的内源性过表达IL-6的人卵巢癌A2780细胞株和内源性抑制IL-6表达的人卵巢癌SKOV-3细胞株,分别采用MTT法、双层软琼脂集落形成实验、流式细胞术、RT-PCR和Western blot技术检测IL-6对OVCA增殖及锚定非依赖生长能力的影响,并对其作用机制进行研究。结果与对照组细胞相比,过表达IL-6的A2780细胞的增殖及锚定非依赖性生长能力增强(P<0.01),而IL-6抑制表达的SKOV-3细胞则降低(P<0.01)。进一步研究表明IL-6促OVCA细胞增殖作用是通过改变细胞周期分布而非抑制细胞凋亡来实现的;过表达IL-6可显著提高A2780细胞的Cyclin D1、Cyclin B1的表达水平,而抑制IL-6表达则明显降低SKOV-3细胞的Cyclin D1、Cyclin B1的表达水平;应用ERK或Akt特异性信号阻断剂可显著抑制高表达IL-6的A2780细胞增殖及Cyclin D1、Cyclin B1的表达水平。结论 IL-6能促进OVCA细胞的增殖和锚定非依赖性生长,IL-6促OVCA细胞增殖作用是通过增加Cyclin D1、Cyclin B1表达水平从而改变细胞周期分布来实现的,并与活化的PI3K/Akt、Ras/MEK/ERK信号通路有关。This study aimed to investigate the effects of endogenous interleukin-6（IL-6） on the proliferation and anchorage-independent growth of ovarian cancer（OVCA） cells in vitro and its mechanism. Based on our previous studies, MTT assay, two layer soft-agar colony formation assay, flow cytometric assay, RT-PCR test and Western blot analysis were used to detect the ability of proliferation and anchorage-independent growth of OVCA A2780 cells overexpressing endogenous IL-6 and IL-6 depleted SKOV-3 cells. We found that the ability of proliferation and anchorage-independent growth were promoted in A2780 cells overexpressing IL-6（P 0.01）, while the ability above were suppressed in IL-6 depleted SKOV-3 cells（P 0.01） compared with the respective controls.Further investigation indicated that IL-6 promoted the proliferation of OVCA cells by altering the distribution of cell cycle rather than inhibiting apoptosis. Overexpressing IL-6 improved significantly the expression of Cyclin D1 and Cyclin B1 in A2780 cells（P 0.05）, while depletion of IL-6 decreased the expression of Cyclin D1 and Cyclin B1 in SKOV-3 cells（P 0.05）. Treatment with specific inhibitor of ERK or Akt signaling pathway inhibited significantly the proliferation and the expression of Cyclin D1 and Cyclin B1 in high IL-6-expressing A2780 cells（P 0.05）. These data suggest that IL-6 can promote proliferation and anchorage-independent growth of OVCA cells. This action of IL- 6 is accomplished by elevating levels of Cyclin D1 and Cyclin B1 and thus altering the distribution of cell cycle, and correlates with activated PI3K/Akt and Ras/MEK/ERK signaling pathways.

关 键 词：IL-6 细胞增殖 锚定非依赖性生长 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]