目的探讨牙龈卟啉单胞菌(Pg)感染对食管鳞癌细胞(ESCC)内IFNGR1棕榈酰化位点突变的机制及临床意义。方法Western blot检测Pg感染不同时间(24、48 h)ESCC细胞KYSE30及KYSE70中IFNGR1蛋白量表达水平。2-BP检测IFNGR1的棕榈酰化作用。构建I...目的探讨牙龈卟啉单胞菌(Pg)感染对食管鳞癌细胞(ESCC)内IFNGR1棕榈酰化位点突变的机制及临床意义。方法Western blot检测Pg感染不同时间(24、48 h)ESCC细胞KYSE30及KYSE70中IFNGR1蛋白量表达水平。2-BP检测IFNGR1的棕榈酰化作用。构建IFNGR1-WT和IFNGR1-C122A棕榈酰化位点突变位点质粒,建立IFNGR1-WT和IFNGR1-C122A稳定细胞系,分别用Pg感染IFNGR1-WT和IFNGR1-C122A细胞,并分为4组:IFNGR1-WT、IFNGR1-C122A、IFNGR1-WT+Pg和IFNGR1-C122A+Pg组。免疫荧光和click-it实验检测IFNGR1在122位点发生棕榈酰化,以及Pg促进IFNGR1在ESCC内发生棕榈酰化。利用平板克隆、划痕实验及Transwell法检测Pg感染前后,IFNGR1-WT和IFNGR1-C122A稳转细胞株的体外增殖、迁移与侵袭能力差异。免疫荧光实验检测IFNGR1与溶酶体标记物LAMP2共定位情况。免疫组化检测50例ESCC组织中Pg感染与IFNGR1蛋白的表达情况;采用卡方检验及R Studio软件分析二者与ESCC患者临床病理特征及生存预后之间的相关性。结果Pg感染ESCC下调IFNGR1的蛋白表达。IFNGR1在122位点发生棕榈酰化。Pg促进IFNGR1在ESCC内棕榈酰化。与IFNGR1-WT组相比,IFNGR1-WT+Pg组体外增殖253±6.245 vs 52±2.45、迁移(0.7816±0.0071)%vs(0.4347±0.0366)%及侵袭709.33±14.57 vs 356.3±17.39能力均显著增强(P<0.05);与IFNGR1-C122A组相比,IFNGR1-C122A+Pg组细胞体外增殖137.33±4.726 vs 29.67±3.055、迁移(0.7477±0.0057)%vs(0.2406±0.0028)%及侵袭587.33±5.033 vs 67.33±2.517能力同样均显著增强(P<0.05)。与IFNGR1-WT+Pg组相比,IFNGR1-C122A+Pg体外增殖137.33±4.726 vs 253±6.245、迁移(0.7477±0.0057)%vs(0.7816±0.0071)%及侵袭587.33±5.033 vs 709.33±14.57能力均显著降低(P<0.05)。Pg及ZDHHC3促进IFNGR1在溶酶体内降解;IFNGR1与Pg表达呈负性相关,且IFNGR1的表达降低与更差的病理特征及生存预后相关。结论Pg感染ESCC下调IFNGR1蛋白表达量,同时诱导IFNGR1棕榈酰化,促进ESCC恶性化进展;IFNGR1的棕榈酰化位点突变减少ESCC增殖、迁移和侵袭作用;Pg感染ESCC靶向IFNGR1到溶酶体内降解可能是由于棕榈酰化作用,因此,清除Pg或抑制棕榈酰化作用能有效抑制食管癌的恶性进展。展开更多
Interferon-gamma(IFN-γ)plays a dual role in cancer;it is both a pro-and an antitumorigenic cytokine,depending on the type of cancer.The deregulation of the IFN-γcanonic pathway is associated with several disorders,i...Interferon-gamma(IFN-γ)plays a dual role in cancer;it is both a pro-and an antitumorigenic cytokine,depending on the type of cancer.The deregulation of the IFN-γcanonic pathway is associated with several disorders,including vulner-ability to viral infections,inflammation,and cancer progression.In particular,the interplay between lung adenocarcinoma(LUAD)and viral infections appears to exist in association with the deregulation of IFN-γsignaling.In this mini-review,we investigated the status of the IFN-γsignaling pathway and the expression level of its components in LUAD.Interestingly,a reduction in IFNGR1 expression seems to be associated with LUAD progression,affecting defenses against viruses such as severe acute respiratory syndrome coronavirus 2.In addition,alterations in the expression of IFNGR1 may inhibit the antiproliferative action of IFN-γsignaling in LUAD.展开更多
文摘目的探讨牙龈卟啉单胞菌(Pg)感染对食管鳞癌细胞(ESCC)内IFNGR1棕榈酰化位点突变的机制及临床意义。方法Western blot检测Pg感染不同时间(24、48 h)ESCC细胞KYSE30及KYSE70中IFNGR1蛋白量表达水平。2-BP检测IFNGR1的棕榈酰化作用。构建IFNGR1-WT和IFNGR1-C122A棕榈酰化位点突变位点质粒,建立IFNGR1-WT和IFNGR1-C122A稳定细胞系,分别用Pg感染IFNGR1-WT和IFNGR1-C122A细胞,并分为4组:IFNGR1-WT、IFNGR1-C122A、IFNGR1-WT+Pg和IFNGR1-C122A+Pg组。免疫荧光和click-it实验检测IFNGR1在122位点发生棕榈酰化,以及Pg促进IFNGR1在ESCC内发生棕榈酰化。利用平板克隆、划痕实验及Transwell法检测Pg感染前后,IFNGR1-WT和IFNGR1-C122A稳转细胞株的体外增殖、迁移与侵袭能力差异。免疫荧光实验检测IFNGR1与溶酶体标记物LAMP2共定位情况。免疫组化检测50例ESCC组织中Pg感染与IFNGR1蛋白的表达情况;采用卡方检验及R Studio软件分析二者与ESCC患者临床病理特征及生存预后之间的相关性。结果Pg感染ESCC下调IFNGR1的蛋白表达。IFNGR1在122位点发生棕榈酰化。Pg促进IFNGR1在ESCC内棕榈酰化。与IFNGR1-WT组相比,IFNGR1-WT+Pg组体外增殖253±6.245 vs 52±2.45、迁移(0.7816±0.0071)%vs(0.4347±0.0366)%及侵袭709.33±14.57 vs 356.3±17.39能力均显著增强(P<0.05);与IFNGR1-C122A组相比,IFNGR1-C122A+Pg组细胞体外增殖137.33±4.726 vs 29.67±3.055、迁移(0.7477±0.0057)%vs(0.2406±0.0028)%及侵袭587.33±5.033 vs 67.33±2.517能力同样均显著增强(P<0.05)。与IFNGR1-WT+Pg组相比,IFNGR1-C122A+Pg体外增殖137.33±4.726 vs 253±6.245、迁移(0.7477±0.0057)%vs(0.7816±0.0071)%及侵袭587.33±5.033 vs 709.33±14.57能力均显著降低(P<0.05)。Pg及ZDHHC3促进IFNGR1在溶酶体内降解;IFNGR1与Pg表达呈负性相关,且IFNGR1的表达降低与更差的病理特征及生存预后相关。结论Pg感染ESCC下调IFNGR1蛋白表达量,同时诱导IFNGR1棕榈酰化,促进ESCC恶性化进展;IFNGR1的棕榈酰化位点突变减少ESCC增殖、迁移和侵袭作用;Pg感染ESCC靶向IFNGR1到溶酶体内降解可能是由于棕榈酰化作用,因此,清除Pg或抑制棕榈酰化作用能有效抑制食管癌的恶性进展。
文摘Interferon-gamma(IFN-γ)plays a dual role in cancer;it is both a pro-and an antitumorigenic cytokine,depending on the type of cancer.The deregulation of the IFN-γcanonic pathway is associated with several disorders,including vulner-ability to viral infections,inflammation,and cancer progression.In particular,the interplay between lung adenocarcinoma(LUAD)and viral infections appears to exist in association with the deregulation of IFN-γsignaling.In this mini-review,we investigated the status of the IFN-γsignaling pathway and the expression level of its components in LUAD.Interestingly,a reduction in IFNGR1 expression seems to be associated with LUAD progression,affecting defenses against viruses such as severe acute respiratory syndrome coronavirus 2.In addition,alterations in the expression of IFNGR1 may inhibit the antiproliferative action of IFN-γsignaling in LUAD.
