前列腺素D2调控自噬影响胃癌干细胞的干性  

Prostaglandin D2 regulation of autophagy affects stemness of gastric cancer stem cells

作　　者：田恒金 汪非凡 高培垚 陈阿敏 王娜 张强 TIAN Hengjin;WANG Feifan;GAO Peiyao;CHEN Amin;WANG Na;ZHANG Qiang(Department of Clinical Laboratory,the First Affiliated Hospital of Bengbu Medical University,Bengbu 233004,China;Department of Blood Transfusion,the First Affiliated Hospital of Naval Medical University,Bengbu 233004,China;Key Laboratory of Cancer Research and Clinical Laboratory Diagnosis,Bengbu Medical University,Bengbu 233004,China)

机构地区：[1]蚌埠医科大学第一附属医院检验科,安徽蚌埠233004 [2]海军军医大学第一附属医院输血科,安徽蚌埠233004 [3]蚌埠医科大学肿瘤基础研究与临床检验诊断重点实验室,安徽蚌埠233004

出　　处：《海南医学院学报》2024年第4期255-261,共7页Journal of Hainan Medical University

基　　金：安徽省自然科学基金面上项目(1908085MH258);蚌埠医学院科研创新项目(Byycxz21004)。

摘　　要：目的:探究前列腺素D2(Prostaglandins D2,PGD2)对胃癌干细胞(gastric cancer stem cells,GCSCs)干性的影响及机制。方法:通过无血清培养法富集7901‐GCSCs;然后通过流式细胞术检测无血清培养的7901‐GCSCs干性标志物CD44的阳性率;通过干细胞成球实验检测不同浓度PGD2(2.5、5、10)μg/mL刺激后的成球能力,并采用Western blot实验检测不同浓度PGD2刺激后干性相关指标(OCT4、CD44)和自噬相关蛋白(LC3、Beclin‐1)的表达。采用Western blot实验检测不同浓度CQ(2.5、5、10)μmol/L刺激后自噬相关蛋白的表达。通过Western blot实验检测PGD2以及PGD2+CQ处理后自噬相关蛋白(LC3、Beclin‐1)和干性相关指标(OCT4、CD44)的蛋白表达。结果:流式细胞术结果显示,与胃癌细胞SGC‐7901相比,无血清培养的7901‐GCSCs中CD44阳性率表达增加(P<0.05)。干细胞成球实验结果表明,与DMSO组相比,PGD2组中7901‐GCSCs的成球能力明显减弱(P<0.05)。Western blot结果显示,与DMSO组相比,PGD2组中7901‐GCSCs的干性相关指标(OCT4、CD44)蛋白表达下调(P<0.05),而自噬相关蛋白(LC3、Beclin‐1)表达增加(P<0.05)。与DMSO组相比,CQ组中自噬相关蛋白(LC3、Beclin‐1)表达降低(P<0.05)。Western blot结果还显示,与DMSO组相比,PGD2+CQ组中细胞自噬相关蛋白以及干性相关指标表达并无明显变化,提示CQ可阻断PGD2对7901‐GCSCs的干性抑制作用。结论:PGD2可能通过调控自噬影响7901‐GCSCs的干性。Objective:To explore the effect and mechanism of prostaglandins D2(PGD2)on the stemness of gastric cancer stem cells(GCSCs).Methods:7901‐GCSCs were enriched by serum‐free culture method;then the positivity rate of CD44,a stemness marker,was detected by flow cytometry in serum‐free cultured 7901‐GCSCs;the sphere‐forming ability was detected by the sphere‐forming assay after stimulation with different concentrations of PGD2(2.5,5,10)μg/mL,and the expression of stemness‐related indicators(OCT4,CD44)and autophagy‐related proteins(LC3,Beclin‐1)after PGD2 stimulation was detected by the Western blot assay in different concentrations.The expression of stemness‐related indexes(OCT4,CD44)and autophagy‐re‐lated proteins(LC3,Beclin‐1)were detected by Western blot assay after stimulation with different concentrations of PGD2.The expression of autophagy‐related proteins after stimulation with different concentrations of CQ(2.5,5,10)μM was detected by Western blot experiment.The protein expression of autophagy‐related proteins(LC3,Beclin‐1)and stemness‐related indexes(OCT4,CD44)was detected by Western blot experiment after PGD2 as well as PGD2+CQ treatment.Results:Flow cytometry results showed that the expression of CD44 positivity was increased in serum‐free cultured 7901‐GCSCs compared with gastric cancer cells SGC‐7901(P<0.05),which fulfilled the needs of subsequent experiments.The results of stem cell spheroid formation assay showed that the spheroid formation ability of 7901‐GCSCs in the PGD2 group was significantly weakened compared with that of the DMSO group(P<0.05).Western blot results showed that the protein expression of stemness‐related indexes(OCT4,CD44)was down‐regulated in the 7901‐GCSCs in the PGD2 group compared with that of the DMSO group(P<0.05),and the expression of autophagy‐related proteins(LC3,Beclin‐1)expression increased(P<0.05).Compared with the DMSO group,the expression of autophagy‐related proteins(LC3,Beclin‐1)was decreased in the CQ group(P

关 键 词：PGD2 胃癌 胃癌干细胞 自噬 干性 

分 类 号：R735.2[医药卫生—肿瘤]