摘要
目的:探讨基质细胞来源因子1(SDF-1)在卵巢癌中的表达情况和对卵巢癌细胞Ca OV3增殖耐药的影响。方法:用Kmplot在线软件分析SDF-1表达水平对卵巢癌患者预后的影响;20μg/ml SDF-1预处理Ca OV3细胞12 h后,在对照组和SDF-1预处理组细胞中克隆形成试验比较细胞克隆形成能力改变;CCk8试验检测细胞增殖能力活性;不同浓度c DDP作用后CCk8试验检测细胞相对活性;c DDP 40μmol/L作用12 h后荧光共聚焦confocal检测细胞DNA双链损伤标志蛋白γ-H2AX表达及同源重组修复关键蛋白RAD51表达,以及两者细胞核中共定位;Western blotting法比较分离得到的配对正常卵巢成纤维细胞NOF和肿瘤相关成纤维细胞中SDF-1的表达情况。结果:在2个卵巢癌公共数据库GSE26712(HR=1.82,P=0.000 21)和GSE9891(HR=1.73,P=0.000 56)中SDF-1高表达组预后显著差于低表达组;第7天对照组和SDF-1组细胞克隆数目分别为(52.3±6.2)和(135.2±8.4);第14天分别为(114.3±3.8)和(258.2±8.6)(P<0.01);SDF-1组细胞增殖速度明显高于对照组(P<0.01);c DDP 40μmol/L作用48 h对照组和SDF-1组细胞存活率分别为(43.2±3.5)%和(67.8±6.6)%(P<0.01);c DDP 40μmol/L作用12 h SDF-1组γ-H2AX较对照组明显减少,同时DNA损伤部位却募集到更多的RAD51;Western blot结果示配对CAF较NOF表达更高的SDF-1。结论:SDF-1在卵巢癌间质细胞CAF中高表达,促进肿瘤细胞增殖与克隆形成能力,提高细胞同源重组修复能力介导c DDP耐药,其高表达是卵巢癌预后不良的标志。
Objective： To explore the expression of SDF-1 in ovarian cancer and the effect of SDF-1 on cell proliferation and cis- platin sensitivity in ovrian cancer cell line CaOV3. Methods： Online Kmplot software （http： //kmplot. com/ovar） was taken to analyze the effect of SDF-1 on prognosis of patients with ovarian cancer. After 12 h pretreatment with 20 μg/ml SDF-1 in CaOV3 cells, colony formation assay and CCK8 test were performed to inspect the colony formation ability and proliferation potential respectively. Meanwhile, CCK8 was used to detect the cell viability after 48 h treatment of various dosage of cDDP in both groups. Furthermore, confocal was adopted to check the production, as well as the co-localization of DNA double strand breaks marker γ-H2AX and repair key protein RAD51. Western blot was done to compare the protein level of SDF-1 in paired NOF and CAF cells. Results： Higher expression of SDF-1 indicated poorer overall prognosis in two ovarian cancer datasets GSE26712 （ HR= 1.82, P= 0. 000 21 ） and GSE9891 （ HR = 1.73, P = 0. 000 56）. The colony formation number in the control group and SDF-1 group was 52. 3±6. 2 and 135.2±8.4 on day 7 （P〈0. 01 ）, and 114. 3±3.8 and 258.2± 8.6 on day 14 （P〈0. 01 ）. The proliferation rate in the SDF-1 group was obviously higher than that of control group （P〈0. 01 ）. The cell viability was 43. 2±3.5% and 67.8±6.6% in the control group and SDF-1 group after 48 h intervention of 40 μmol/L cDDP （P〈0. 01 ）. Confocal showed that γ-H2AX in the SDF-1 group was generously lower, and RAD51 was notably higher compared with the control group. Western blot confirmed the remarkably higher protein level of SDF- 1 in CAF cells compared with paired NOF cells. Conclusion： SDF - 1 is enriched in ovarian cancer sromal CAF cells, could promote the proliferation and colony formation capacity of ovarian cancer cells, en- hance the homologous repair ability and thus mediate cDDP resistence, and is an independent marker for poor prognosis in
出处
《中国妇幼保健》
CAS
2015年第19期3287-3290,共4页
Maternal and Child Health Care of China
