【摘要】：目的探討胃癌衰老細(xì)胞相關(guān)分泌表型條件培養(yǎng)基(SASP-CM)對(duì)人胃癌細(xì)胞系BGC823增殖能力的影響。方法取BGC823細(xì)胞分為3組:胃癌SASP-CM組、正常腫瘤細(xì)胞條件培養(yǎng)基(CTR-CM)組和正常培養(yǎng)基(NOR-CM)組。利用紫杉醇(PTX)處理BGC823細(xì)胞構(gòu)建衰老細(xì)胞模型并制備SASP-CM。使用β半乳糖苷酶染色驗(yàn)證衰老細(xì)胞模型的建立;酶聯(lián)免疫吸附實(shí)驗(yàn)檢測(cè)SASP-CM中主要的衰老細(xì)胞相關(guān)分泌表型(SASP)因子的濃度;CCK-8法及細(xì)胞克隆形成實(shí)驗(yàn)檢測(cè)SASP-CM對(duì)BGC823細(xì)胞增殖能力的影響;流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期及細(xì)胞凋亡情況。結(jié)果 35nmol/L PTX誘導(dǎo)BGC823細(xì)胞72h可建立穩(wěn)定的細(xì)胞衰老模型,此條件下衰老細(xì)胞比例最高,為(66.95±3.54)%。SASP-CM中主要的SASP因子白細(xì)胞介素(IL)-6、IL-8、CXC趨化因子配體1(CXCL1)、CC趨化因子配體2(CCL2)、γ干擾素(INF-γ)的濃度均高于CTR-CM(P均0.01)。培養(yǎng)48、72、96h時(shí)SASP-CM組細(xì)胞的增殖活性均高于CTR-CM組和NOR-CM組(P均0.05)。SASP-CM組細(xì)胞的相對(duì)克隆形成率高于CTR-CM組(P0.01)和NOR-CM組(P0.05)。SASP-CM組的S期細(xì)胞比例均高于CTR-CM組和NOR-CM組(P均0.01),各組間細(xì)胞凋亡率差異無(wú)統(tǒng)計(jì)學(xué)意義。結(jié)論胃癌SASP-CM促進(jìn)BGC823細(xì)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 鹵3.54)%. The concentrations of (IL)-6, IL 鈮，

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