| 摘要: |
| 目的探讨衰老的卵巢间质成纤维细胞对卵巢癌细胞系SKOV3的增殖、侵袭能力的影响。方法原代培养人卵巢正常成纤维细胞（NOF），在体外持续传代诱导细胞衰老，用衰老相关的β半乳糖苷酶染色（SA-b-gal）试验检测细胞是否发生衰老。收集正常成纤维细胞的条件培养基（CMn）及衰老成纤维细胞的条件培养基（CMs）。用CMn和CMs分别处理SKOV3细胞，利用CCK8试剂盒、Transwell小室及三维培养试验检测细胞的增殖、侵袭能力的变化。结果卵巢正常成纤维细胞在体外持续传代会导致细胞复制性衰老，在SA-b-gal试验中有明显的深蓝色沉淀产物。用CMs处理SKOV3细胞，CCK8试验显示细胞培养第5天的OD（2.067±0.058）高于CMn组（1.073±0.056）（t=21.19，P＜0.01)，表明细胞的增殖能力增强。Transwell试验显示CMs组穿透Matrigel胶的细胞数（235.00±29.68）个，较CMn组的（129.30±15.52）个明显增多（t=5.47，P＜0.01）；三维培养中形成的克隆球CMs组较CMn组明显增大、数量明显增多（27.00±2.16vs11.67±2.50；t=8.04，P＜0.01)，向外侵袭能力增强。结论衰老的卵巢间质成纤维细胞在体外培养中能促进卵巢癌细胞的增殖和侵袭能力。 |
| 关键词: 卵巢癌 衰老 间质成纤维细胞 三维培养 |
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| Senescent ovarian stromal fibroblasts promotes the proliferation and invasion ability of human ovarian cancer cells |
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GAO Wen, ZHU Tao
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Zhejiang Carcer Hospital
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| Abstract: |
| Objective To investigate the effect of senescent ovarian stromal fibroblasts on tumor growth and invasive ability of human ovarian cancer cells. Methods Normal human ovarian fibroblasts(NOF) were primarily cultured and senescence was induced by replicative exhaustion. Senescence associated beta-galactosidase staining was used to detect cell senescence of normal ovarian fibroblasts after serial subcultivation. Normal stromal fibroblasts conditioned mediums (CMn) and senescent stromal fibroblasts conditioned mediums (CMs) were collected. Ovarian cancer SKOV3 cells were treated with CMn or CMs. The cell proliferation and invasion were measured by cell counting kit 8 (CCK8) and Transwell assay and three-dimensional cell culture, respectively. Results Replicative senescence was induced in NOF after serial subcultivation. Beta-galactosidase staining showed blue granules within cytoplasm of the senescent ovarian stromal fibroblasts. CCK8 assay showed that the proliferation of SKOV3 cells in CMs group was markedly increased compared to CMn group (OD value: 2.067±0.058 vs 1.073±
0.056, t=21.19, P＜0.01). Transwell assay that the migration and invasion of SKOV3 cells in CMs group were markedly increased compared with CMn group (235.00±29.68 vs 129.30±15.52, t=5.47, P＜0.01). Three-dimensional culture showed that the clone spheres of SKOV3 cells in CMs group were larger and more aggressive than those in CMn group; and the number of clone spheres in CMS group were significantly increased compared with CMn group (27.00 ±2.16 vs 11.67 ±2.50, t=8.04, P＜0.01). Conclusion Our results indicates that senescent ovarian stromal fibroblasts may enhance proliferation and invasion of human o- varian cancer cells. |
| Key words: Ovarian cancer Senescence Stromal fibroblasts Three-dimensional cell culture |
