海南医学院学报2024，Vol.30Issue(4)：255-261,7.DOI:10.13210/j.cnki.jhmu.20231212.002
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
摘要
Abstract
Objective:To explore the effect and mechanism of prostaglandins D2(PGD2)on the stemness of gastric can-cer 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 stem-ness-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 cytome-try 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 forma-tion 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<0.05).Western blot re-sults also showed that the expression of cellular autophagy-related proteins and stemness-related indexes in the PGD2+CQ group was not significantly changed compared with that of the DMSO group(ns:the difference was not significant),suggesting that the CQ could block the effect of PGD2 on the expression of stemness markers in 7901-GCSCs.Conclusion:PGD2 may affect the stemness of 7901-GCSCs by regulating autophagy.关键词
PGD2/胃癌/胃癌干细胞/自噬/干性Key words
PGD2/Gastric cancer/Gastric cancer stem cells/Autophagy/Stemness分类
医药卫生引用本文复制引用
田恒金,汪非凡,高培垚,陈阿敏,王娜,张强..前列腺素D2调控自噬影响胃癌干细胞的干性[J].海南医学院学报,2024,30(4):255-261,7.基金项目
This study was supported by the Natural Science Foundation of Anhui Province(1908085MH258) （1908085MH258）
Scientific Research and Innovation Project of Bengbu Medical College(Byycxz21004)安徽省自然科学基金面上项目(1908085MH258) （Byycxz21004）
蚌埠医学院科研创新项目(Byycxz21004) （Byycxz21004）
