胃癌衰老细胞相关分泌表型条件培养基对人胃癌细胞系BGC823增殖的影响
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长征医院,长征医院,长征医院,第二军医大学长征医院普外二科

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国家自然科学基金(81372670,81100629,81402359,81602617),上海市自然科学基金(16ZR1436800).


Effect of senescence-associated secretory phenotype-conditioned medium on proliferation of human gastric cancer cell lines BGC823
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Changzhang hospital,,,Changzheng Hospital

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Supported by National Natural Science Foundation of China (81372670, 81100629, 81402359, 81602617) and Natural Science Foundation of Shanghai (16ZR1436800).

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    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。

    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.

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  • 收稿日期:2017-06-19
  • 最后修改日期:2017-10-10
  • 录用日期:2017-10-25
  • 在线发布日期: 2017-12-21
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