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 CD4...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 autophagyrelated 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-related 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<0.05).Western blot results 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.7901-GCSCs stemness inhibition.Conclusion:PGD2 may affect the stemness of 7901-GCSCs by regulating autophagy.展开更多
Objective:To preliminarily investigate the possible role of prostaglandin D_2(PGD_2) in malaria infections.Methods:Blood and urinary samples(n=120 each) were collected from Thai patients with Plasmodium falciparum(P.f...Objective:To preliminarily investigate the possible role of prostaglandin D_2(PGD_2) in malaria infections.Methods:Blood and urinary samples(n=120 each) were collected from Thai patients with Plasmodium falciparum(P.falciparum) with moderate(n=26) and high(n=4) parasitemia,patients with Plasmodium vivax(P.vivax)(n=30),patients with fever associated with other infections(n=30),and healthy subjects(n=30).PGD_2 concentrations in plasma and urinary samples of healthy subjects,patients with fever associated with other infections and patients with malaria were determined using Prostaglandin D2-MOX express EIA kit(Cayman Chemical,USA).Results:The possible association between PGD_2 and malaria infections is clearly demonstrated with PGD_2 concentration in urine.The urinary PGD_2 concentrations were relatively high(about 5-fold) in patients with P.falciparum with moderate parasitemia and P.vivax infections compared with other groups.Furthermore,the concentration in patients with P.falciparum with moderate parasitemia and P.vivax infection were significantly higher than that in healthy subjects and patients with fever associated with other infections.Conclusions:Urinary PGD_2 concentrations may offer a more dependable and useful tool for predicting malaria severity.Confirmation is this preliminary finding is required with a larger sample size.展开更多
基金Natural Science Foundation of Anhui Province(No.1908085MH258)Scientific Research and Innovation Project of Bengbu Medical College(No.Byycxz21004)。
文摘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 autophagyrelated 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-related 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<0.05).Western blot results 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.7901-GCSCs stemness inhibition.Conclusion:PGD2 may affect the stemness of 7901-GCSCs by regulating autophagy.
基金funded by Thailand Research Fund-Thammasat University Joint Fund and Graduated Student Grant to P.Thongdee(No.PHD/0365/2552)
文摘Objective:To preliminarily investigate the possible role of prostaglandin D_2(PGD_2) in malaria infections.Methods:Blood and urinary samples(n=120 each) were collected from Thai patients with Plasmodium falciparum(P.falciparum) with moderate(n=26) and high(n=4) parasitemia,patients with Plasmodium vivax(P.vivax)(n=30),patients with fever associated with other infections(n=30),and healthy subjects(n=30).PGD_2 concentrations in plasma and urinary samples of healthy subjects,patients with fever associated with other infections and patients with malaria were determined using Prostaglandin D2-MOX express EIA kit(Cayman Chemical,USA).Results:The possible association between PGD_2 and malaria infections is clearly demonstrated with PGD_2 concentration in urine.The urinary PGD_2 concentrations were relatively high(about 5-fold) in patients with P.falciparum with moderate parasitemia and P.vivax infections compared with other groups.Furthermore,the concentration in patients with P.falciparum with moderate parasitemia and P.vivax infection were significantly higher than that in healthy subjects and patients with fever associated with other infections.Conclusions:Urinary PGD_2 concentrations may offer a more dependable and useful tool for predicting malaria severity.Confirmation is this preliminary finding is required with a larger sample size.
