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
发布时间：2019-06-30 21:20
【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 35nmol/L PTX诱导BGC823细胞72h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、96h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(P0.01)和NOR-CM组(P0.05)。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 gastric cancer aging cell associated secretory phenotypic conditioned medium (SASP-CM) on the proliferation of human gastric cancer cell line BGC823. Methods BGC823 cells were divided into three groups: gastric cancer SASP-CM group, normal tumor cell conditioned medium (CTR-CM) group and normal culture medium (NOR-CM) group. Establishment of aging cell model and preparation of SASP-CM. in BGC823 cells treated with paclitaxel (PTX) 尾 galactosidase staining was used to verify the establishment of aging cell model, enzyme linked immunosorbent assay (Elisa) was used to detect the concentration of phenotypic (SASP) factors secreted by aging cells in SASP-CM, CCK- 8 assay and cell clone formation assay were used to detect the effect of SASP-CM on the proliferation of BGC823 cells, and flow cytometry was used to detect cell cycle and apoptosis. Results BGC823 cells induced by 35nmol/L PTX for 72 h could establish a stable cell senescence model, and the proportion of senescent cells was the highest (66.95 卤3.54)%. The concentrations of (IL)-6, IL 鈮，
本文编号：2508261
[Abstract]:Objective to investigate the effect of gastric cancer aging cell associated secretory phenotypic conditioned medium (SASP-CM) on the proliferation of human gastric cancer cell line BGC823. Methods BGC823 cells were divided into three groups: gastric cancer SASP-CM group, normal tumor cell conditioned medium (CTR-CM) group and normal culture medium (NOR-CM) group. Establishment of aging cell model and preparation of SASP-CM. in BGC823 cells treated with paclitaxel (PTX) 尾 galactosidase staining was used to verify the establishment of aging cell model, enzyme linked immunosorbent assay (Elisa) was used to detect the concentration of phenotypic (SASP) factors secreted by aging cells in SASP-CM, CCK- 8 assay and cell clone formation assay were used to detect the effect of SASP-CM on the proliferation of BGC823 cells, and flow cytometry was used to detect cell cycle and apoptosis. Results BGC823 cells induced by 35nmol/L PTX for 72 h could establish a stable cell senescence model, and the proportion of senescent cells was the highest (66.95 卤3.54)%. The concentrations of (IL)-6, IL 鈮，
本文编号：2508261
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