目的探究3’-磷酸腺苷-5’-磷酸硫酸盐合酶2(PAPSS2)在胃癌组织和细胞中的表达情况及其对胃癌细胞侵袭和迁移的影响。方法利用GEPIA数据库分析PAPSS2在胃癌组织中的表达情况。收集陕西省人民医院2016年10月至2018年6月40例胃癌手术患者...目的探究3’-磷酸腺苷-5’-磷酸硫酸盐合酶2(PAPSS2)在胃癌组织和细胞中的表达情况及其对胃癌细胞侵袭和迁移的影响。方法利用GEPIA数据库分析PAPSS2在胃癌组织中的表达情况。收集陕西省人民医院2016年10月至2018年6月40例胃癌手术患者的癌组织及癌旁正常组织标本,进一步通过免疫组织化学染色进行验证。分析PAPSS2表达与胃癌患者临床病理资料和预后的关系。利用qPCR和免疫印迹法分别检测PAPSS2在胃癌细胞(HGC-27、SNU-1、AGS、SGC-7901)和正常胃黏膜细胞系(GES-1)的mRNA和蛋白水平表达差异。利用shRNA敲低胃癌细胞PAPSS2的表达,MTT检测其对细胞活性的影响,Transwell检测其对侵袭和迁移能力的影响。结果 (1)利用GEPIA数据库分析发现,PAPSS2在胃癌组织中高表达(P<0.05),且高表达PAPSS2的胃癌患者总体生存率(HR=1.5,P=0.017)和无病生存率(HR=1.6,P=0.014)显著降低。(2)PAPSS2在40例胃癌组织的免疫组织化学染色评分显著高于对应的癌旁组织(7.100±3.169 vs 3.425±2.263,P<0.001)。高表达PAPSS2的胃癌患者肿瘤浸润深度(P=0.015)和淋巴转移率更高(P=0.005)。(3)qPCR显示PAPSS2在胃癌细胞(AGS、HGC-27、SNU-1、SGC-7901)的mRNA表达水平明显高于GES-1细胞(F=15.45,P<0.001),免疫印迹法也得出了类似的结果。(4)MTT实验发现敲低PAPSS2对AGS和SGC-7901细胞的活性没有明显影响。(5)敲低PAPSS2后AGS细胞(386.9±73.75 vs 602.7±79.23,t=4.457,P=0.002)和SGC-7901细胞(237.8±77.60 vs 461.8±86.55,t=4.309,P=0.003)的迁移能力显著降低,AGS细胞(106.0±38.07 vs 201.3±48.79,t=3.446,P=0.009)和SGC-7901细胞(55.0±18.27 vs 93.6±21.07,t=3.093,P=0.015)的侵袭能力也明显降低。结论 PAPSS2在胃癌组织和细胞中呈高表达,其高表达促进了胃癌细胞的侵袭和转移,这可能是胃癌预后较差的重要原因之一。展开更多
Sulfation is a crucial and prevalent conjugation reaction involved in cellular processes and mammalian physiology.3’-Phosphoadenosine 5’-phosphosulfate(PAPS)synthase 2(PAPSS2)is the primary enzyme to generate the un...Sulfation is a crucial and prevalent conjugation reaction involved in cellular processes and mammalian physiology.3’-Phosphoadenosine 5’-phosphosulfate(PAPS)synthase 2(PAPSS2)is the primary enzyme to generate the universal sulfonate donor PAPS.The involvement of PAPSS2-mediated sulfation in adenomatous polyposis coli(APC)mutation-promoted colonic carcinogenesis has not been reported.Here,we showed that the expression of PAPSS2 was decreased in human colon tumors along with cancer stages,and the lower expression of PAPSS2 was correlated with poor prognosis in advanced colon cancer.Gut epithelial-specific heterozygous Apc deficient and Papss2-knockout(Apc^(Δgut-Het)Papss2^(Δgut))mice were created,and the phenotypes were compared to the spontaneous intestinal tumorigenesis of Apc^(Δgut-Het)mice.Apc^(Δgut-Het)Papss2^(Δgutmice) were more sensitive to gut tumorigenesis,which was mechanistically accounted for by the activation of Wnt/β-catenin signaling pathway due to the suppression of chondroitin sulfation and inhibition of the farnesoid X receptor(FXR)-transducin-like enhancer of split 3(TLE3)gene regulatory axis.Chondroitin sulfate supplementation in Apc^(Δgut-Het)Papss2^(Δgutmice) alleviated intestinal tumorigenesis.In summary,we have uncovered the protective role of PAPSS2-mediated chondroitin sulfation and bile acids-FXR-TLE3 activation in the prevention of gut carcinogenesis via the antagonization of Wnt/β-catenin signaling.Chondroitin sulfate may be explored as a therapeutic agent for Papss2 deficiency-associated colonic carcinogenesis.展开更多
文摘目的探究3’-磷酸腺苷-5’-磷酸硫酸盐合酶2(PAPSS2)在胃癌组织和细胞中的表达情况及其对胃癌细胞侵袭和迁移的影响。方法利用GEPIA数据库分析PAPSS2在胃癌组织中的表达情况。收集陕西省人民医院2016年10月至2018年6月40例胃癌手术患者的癌组织及癌旁正常组织标本,进一步通过免疫组织化学染色进行验证。分析PAPSS2表达与胃癌患者临床病理资料和预后的关系。利用qPCR和免疫印迹法分别检测PAPSS2在胃癌细胞(HGC-27、SNU-1、AGS、SGC-7901)和正常胃黏膜细胞系(GES-1)的mRNA和蛋白水平表达差异。利用shRNA敲低胃癌细胞PAPSS2的表达,MTT检测其对细胞活性的影响,Transwell检测其对侵袭和迁移能力的影响。结果 (1)利用GEPIA数据库分析发现,PAPSS2在胃癌组织中高表达(P<0.05),且高表达PAPSS2的胃癌患者总体生存率(HR=1.5,P=0.017)和无病生存率(HR=1.6,P=0.014)显著降低。(2)PAPSS2在40例胃癌组织的免疫组织化学染色评分显著高于对应的癌旁组织(7.100±3.169 vs 3.425±2.263,P<0.001)。高表达PAPSS2的胃癌患者肿瘤浸润深度(P=0.015)和淋巴转移率更高(P=0.005)。(3)qPCR显示PAPSS2在胃癌细胞(AGS、HGC-27、SNU-1、SGC-7901)的mRNA表达水平明显高于GES-1细胞(F=15.45,P<0.001),免疫印迹法也得出了类似的结果。(4)MTT实验发现敲低PAPSS2对AGS和SGC-7901细胞的活性没有明显影响。(5)敲低PAPSS2后AGS细胞(386.9±73.75 vs 602.7±79.23,t=4.457,P=0.002)和SGC-7901细胞(237.8±77.60 vs 461.8±86.55,t=4.309,P=0.003)的迁移能力显著降低,AGS细胞(106.0±38.07 vs 201.3±48.79,t=3.446,P=0.009)和SGC-7901细胞(55.0±18.27 vs 93.6±21.07,t=3.093,P=0.015)的侵袭能力也明显降低。结论 PAPSS2在胃癌组织和细胞中呈高表达,其高表达促进了胃癌细胞的侵袭和转移,这可能是胃癌预后较差的重要原因之一。
基金supported by NIH grants DK117370,DK135538,and ES030429(to Wen Xie,US)a Pilot&Feasibility grant(to Pengfei Xu,US)from the Pittsburgh Liver Research Center funded by NIH grant P30DK120531NIH shared instrumentation grant:Olympus FV3000 Confocal Microscope SIG:NIH S10OD030254-01A1。
文摘Sulfation is a crucial and prevalent conjugation reaction involved in cellular processes and mammalian physiology.3’-Phosphoadenosine 5’-phosphosulfate(PAPS)synthase 2(PAPSS2)is the primary enzyme to generate the universal sulfonate donor PAPS.The involvement of PAPSS2-mediated sulfation in adenomatous polyposis coli(APC)mutation-promoted colonic carcinogenesis has not been reported.Here,we showed that the expression of PAPSS2 was decreased in human colon tumors along with cancer stages,and the lower expression of PAPSS2 was correlated with poor prognosis in advanced colon cancer.Gut epithelial-specific heterozygous Apc deficient and Papss2-knockout(Apc^(Δgut-Het)Papss2^(Δgut))mice were created,and the phenotypes were compared to the spontaneous intestinal tumorigenesis of Apc^(Δgut-Het)mice.Apc^(Δgut-Het)Papss2^(Δgutmice) were more sensitive to gut tumorigenesis,which was mechanistically accounted for by the activation of Wnt/β-catenin signaling pathway due to the suppression of chondroitin sulfation and inhibition of the farnesoid X receptor(FXR)-transducin-like enhancer of split 3(TLE3)gene regulatory axis.Chondroitin sulfate supplementation in Apc^(Δgut-Het)Papss2^(Δgutmice) alleviated intestinal tumorigenesis.In summary,we have uncovered the protective role of PAPSS2-mediated chondroitin sulfation and bile acids-FXR-TLE3 activation in the prevention of gut carcinogenesis via the antagonization of Wnt/β-catenin signaling.Chondroitin sulfate may be explored as a therapeutic agent for Papss2 deficiency-associated colonic carcinogenesis.
