| 摘要: |
| 目的 探讨胃癌衰老细胞相关分泌表型条件培养基（SASP-CM）对人胃癌细胞系BGC823增殖能力的影响。方法 取BGC823细胞分为3组：胃癌SASP-CM组、正常肿瘤细胞条件培养基（CTR-CM）组和正常培养基（NOR-CM）组。利用紫杉醇（PTX）处理BGC823细胞构建衰老细胞模型并制备SASP-CM。使用β半乳糖苷酶染色验证衰老细胞模型的建立；酶联免疫吸附实验检测SASP-CM中主要的衰老细胞相关分泌表型（SASP）因子的浓度；CCK-8法及细胞克隆形成实验检测SASP-CM对BGC823细胞增殖能力的影响；流式细胞术检测细胞周期及细胞凋亡情况。结果 35 nmol/L PTX诱导BGC823细胞72 h可建立稳定的细胞衰老模型，此条件下衰老细胞比例最高，为（66.95±3.54）%。SASP-CM中主要的SASP因子白细胞介素（IL）-6、IL-8、CXC趋化因子配体1（CXCL1）、CC趋化因子配体2（CCL2）、γ干扰素（INF-γ）的浓度均高于CTR-CM （P均<0.01）。培养48、72、96 h时SASP-CM组细胞的增殖活性均高于CTR-CM组和NOR-CM组（P均<0.05）。SASP-CM组细胞的相对克隆形成率高于CTR-CM组（P<0.01）和NOR-CM组（P<0.05）。SASP-CM组的S期细胞比例均高于CTR-CM组和NOR-CM组（P均<0.01），各组间细胞凋亡率差异无统计学意义。结论 胃癌SASP-CM促进BGC823细胞增殖。 |
| 关键词: 胃癌 细胞衰老 衰老相关分泌表型 紫杉醇 细胞增殖 |
| DOI：10.16781/j.0258-879x.2017.12.1508 |
| 投稿时间：2017-06-19修订日期：2017-10-10 |
| 基金项目:国家自然科学基金（81372670，81100629，81402359，81602617），上海市自然科学基金（16ZR1436800）. |
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| Effect of senescence-associated secretory phenotype-conditioned medium on proliferation of human gastric cancer cell lines BGC823 |
| WANG Yu△,ZHU Zhen-xin△,TANG Yuan,FENG Dan,ZHANG Xin,WANG Chang-ming,CAI Qing-ping* |
(Department of General Surgery(Ⅱ), Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
△Co-first authors.
* Corresponding author) |
| Abstract: |
| Objective To investigate the effect of senescence-associated secretory phenotype-conditioned medium (SASP-CM) on the proliferation of human gastric cancer cell lines BGC823. Methods BGC823 cells were divided into three groups:SASP-CM group, tumor cells control-conditioned medium (CTR-CM) group, and normal-conditioned medium (NOR-CM) group. BGC823 cells in the SASP-CM group were treated with paclitaxel (PTX) to establish senescent cell model and to prepare SASP-CM. The establishment of senescent cell model was confirmed by senescence-associated β-galactosidase staining. The concentrations of major SASP factors in SASP-CM were detected by enzyme linked immunosorbent assay, the proliferation ability of BGC823 cells was detected by CCK-8 assay and cell clone formation assay, and the cell cycle and apoptosis were detected by flow cytometry. Results After treating BGC823 cells with 35 nmol/L PTX for 72 h, a steady senescence model was established, which had the highest percentage of senescence cells ([66.95±3.54]%). The concentrations of interleukin (IL)-6, IL-8, chemokine (C-X-C motif) ligand 1 (CXCL1), chemokine (C-C motif) ligand 2 (CCL2), and interferon γ (INF-γ) in the SASP-CM were significantly higher than those in the CTR-CM (all P<0.01). The relative clone formation rate of BGC823 cells in the SASP-CM group was significantly higher than those in the CTR-CM group (P<0.01) and NOR-CM group (P<0.05). The proliferation activity of BGC823 cells in the SASP-CM group was significantly higher than that in the CTR-CM and NOR-CM groups after incubation for 48, 72 and 96 h (all P<0.05). The percentage of S phase cells in the SASP-CM group was significantly higher than that in the CTR-CM and NOR-CM groups (both P<0.01), and there was no significant difference in cell apoptosis rate between groups. Conclusion SASP-CM can promote the proliferation of human gastric cancer cell lines BGC823. |
| Key words: gastric carcinoma cell senescence senescence-associated secretory phenotype taxol cell proliferation |
