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  

Effect of senescence-associated secretory phenotype-conditioned medium on proliferation of human gastric cancer cell lines BGC823

作　　者：王瑜[1] 朱振新[1] 唐渊 冯丹[1] 张鑫[1] 王长明[1] 蔡清萍[1] 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)

机构地区：[1]第二军医大学长征医院普外二科,上海200003

出　　处：《第二军医大学学报》2017年第12期1508-1513,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81372670;81100629;81402359;81602617);上海市自然科学基金(16ZR1436800)~~

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(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。ObjectiveTo investigate the effect of senescence associated secretory phenotype conditioned medium(SASP CM)on the proliferation of human gastric cancer cell lines BGC823.MethodsBGC823cells were divided into three groups:SASP CM group,tumor cells control conditioned medium(CTR CM)group,and normal conditioned medium(NOR CM)group.BGC823cells 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 associatedβgalactosidase staining.The concentrations of major SASP factors in SASP CM were detected by enzyme linked immunosorbent assay,the proliferation ability of BGC823cells was detected by CCK8assay and cell clone formation assay,and the cell cycle and apoptosis were detected by flow cytometry.ResultsAfter treating BGC823cells with35nmol/L PTX for72h,a steady senescence model was established,which had the highest percentage of senescence cells([66.95±3.54]%).The concentrations of interleukin(IL)6,IL8,chemokine(C X C motif)ligand1(CXCL1),chemokine(C C motif)ligand2(CCL2),and interferonγ(INFγ)in the SASP CM were significantly higher than those in the CTR CM(all P<0.01).The relative clone formation rate of BGC823cells 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 BGC823cells in the SASP CM group was significantly higher than that in the CTR CM and NOR CM groups after incubation for48,72and96h(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.ConclusionSASP CM can promote the proliferation of human gastric cancer cell lines BGC823.

关 键 词：胃癌 细胞衰老 衰老相关分泌表型 紫杉醇 细胞增殖 

分 类 号：R364.33[医药卫生—病理学]