Colorectal cancer(CRC)ranks among the top five most common malignant tumors worldwide and has a high mortality rate.Angiogenesis plays an important role in CRC progression;however,anti-angiogenesis therapy still has m...Colorectal cancer(CRC)ranks among the top five most common malignant tumors worldwide and has a high mortality rate.Angiogenesis plays an important role in CRC progression;however,anti-angiogenesis therapy still has many limitations.Long non-coding RNAs(lncRNAs)participate in tumor progression by regulating the expression of vascular endothelial growth factor in metastatic CRC.Thus,targeting specific lncRNAs may provide some new hope for anti-angiogenic strategies.Through analyzing data from both clinical samples and The Cancer Genome Atlas database,we found that the lncRNA LINC01503 was specifically upregulated in CRC tissues and was associated with tumor progression and poor overall survival.We also demonstrated that LINC01503 enhanced the capacity for tube formation and migration of vascular endothelial cells,thus promoting CRC tumorigenesis by upregulating vascular endothelial growth factor A(VEGFA)expression in CRC cells.Mechanistically,LINC01503 promoted the expression of VEGFA by simultaneously regulating both mRNA and protein stability of VEGFA by binding to miR-342-3p and the chaperone HSP60,respectively.The upregulation of LINC01503 in CRC cells was attributed to the CREB-binding protein CBP/p300-mediated H3K27 acetylation of the LINC01503 promoter region.Taken together,our findings clarify the mechanism by which LINC01503 may promote CRC angiogenesis,implying that LINC01503 may serve as a potential prognostic biomarker and therapeutic target for CRC.展开更多
BACKGROUND Cervical cancer(CC)stem cell-like cells(CCSLCs),defined by the capacity of differentiation and self-renewal and proliferation,play a significant role in the progression of CC.However,the molecular mechanism...BACKGROUND Cervical cancer(CC)stem cell-like cells(CCSLCs),defined by the capacity of differentiation and self-renewal and proliferation,play a significant role in the progression of CC.However,the molecular mechanisms regulating their self-renewal are poorly understood.Therefore,elucidation of the epigenetic mechanisms that drive cancer stem cell self-renewal will enhance our ability to improve the effectiveness of targeted therapies for cancer stem cells.AIM To explore how DNA methyltransferase 1(DNMT1)/miR-342-3p/Forkhead box M1(FoxM1),which have been shown to have abnormal expression in CCSLCs,and their signaling pathways could stimulate self-renewal-related stemness in CCSLCs.METHODS Sphere-forming cells derived from CC cell lines HeLa,SiHa and CaSki served as CCSLCs.Self-renewal-related stemness was identified by determining sphere and colony formation efficiency,CD133 and CD49f protein level,and SRY-box transcription factor 2 and octamer-binding transcription factor 4 mRNA level.The microRNA expression profiles between HeLa cells and HeLa-derived CCSLCs or mRNA expression profiles that HeLaderived CCSLCs were transfected with or without miR-342-3p mimic were compared using quantitative PCR analysis.The expression levels of DNMT1 mRNA,miR-342-3p,and FoxM1 protein were examined by quantitative real-time PCR and western blotting.In vivo carcinogenicity was assessed using a mouse xenograft model.The functional effects of the DNMT1/miR-342-3p/FoxM1 axis were examined by in vivo and in vitro gain-of-activity and loss-of-activity assessments.Interplay among DNMT1,miR-342-3p,and FoxM1 was tested by methylationspecific PCR and a respective luciferase reporter assay.RESULTS CCSLCs derived from the established HeLa cell lines displayed higher self-renewal-related stemness,including enhanced sphere and colony formation efficiency,increased CD133 and CD49f protein level,and heightened transcriptional quantity of stemness-related factors SRY-box transcription factor 2 and octamer-binding transcription factor 4 in vitro as well as a stronger tumorigenic potential in vivo compared to their parental cells.Moreover,quantitative PCR showed that the miR-342-3p level was downregulated in HeLa-derived CCSLCs compared to HeLa cells.Its mimic significantly decreased DNMT1 and FoxM1 mRNA expression levels in CCSLCs.Knockdown of DNMT1 or miR-342-3p mimic transfection suppressed DNMT1 expression,increased miR-342-3p quantity by promoter demethylation,and inhibited CCSLC self-renewal.Inhibition of FoxM1 by shRNA transfection also resulted in the attenuation of CCSLC self-renewal but had little effect on the DNMT1 activity and miR-342-3p expression.Furthermore,the loss of CCSLC self-renewal exerted by miR-342-3p mimic was inverted by the overexpression of DNMT1 or FoxM1.Furthermore,DNMT1 and FoxM1 were recognized as straight targets by miR-342-3p in HeLa-derived CCSLCs.CONCLUSION Our findings suggested that a novel DNMT1/miR-342-3p/FoxM1 signal axis promotes CCSLC self-renewal and presented a potential target for the treatment of CC through suppression of CCSLC self-renewal.However,this pathway has been previously implicated in CC,as evidenced by prior studies showing miR-342-3p-mediated downregulation of FoxM1 in cervical cancer cells.Additionally,research on liver cancer further supports the involvement of miR-342-3p in suppressing FoxM1 expression.While our study contributed to this body of knowledge,we did not present a completely novel axis but reinforced the therapeutic potential of targeting the DNMT1/miR-342-3p/FoxM1 axis to suppress CCSLC self-renewal in CC treatment.展开更多
目的:探讨miR-342-3p在鼻咽癌(NPC)中的表达及其意义。方法:miRNA芯片检测人永生化鼻咽上皮细胞系NP69和低分化NPC细胞CNE2中miRNA的差异表达;实时荧光定量PCR(qPCR)法检测NPC组织中miR-342-3p的表达;GO and Pathway分析miR-342-3p在NP...目的:探讨miR-342-3p在鼻咽癌(NPC)中的表达及其意义。方法:miRNA芯片检测人永生化鼻咽上皮细胞系NP69和低分化NPC细胞CNE2中miRNA的差异表达;实时荧光定量PCR(qPCR)法检测NPC组织中miR-342-3p的表达;GO and Pathway分析miR-342-3p在NPC发生发展中可能涉及的功能及其参与的信号通路。结果:miRNA芯片结果表明,miR-342-3p在NPC细胞CNE2中低表达,qPCR结果进一步证实miR-342-3p在NPC组织中低表达;GO分析表明,具有显著差异的10个功能中,与NPC发生发展关系较为密切的是miR-342-3p。Pathway分析显示,miR-342-3p与丝裂原应激活化蛋白(MAPK)信号通路相关性较大。结论:miR-342-3p的低表达可能与NPC的发生和发展相关。展开更多
基金supported by the National Natural Science Foundation of China(Grant No.81972288)the Scientific and Technologic Development Programme of Suzhou(Livelihood Science and Technology-Applied Basic Research in Healthcare,SYS2020057).
文摘Colorectal cancer(CRC)ranks among the top five most common malignant tumors worldwide and has a high mortality rate.Angiogenesis plays an important role in CRC progression;however,anti-angiogenesis therapy still has many limitations.Long non-coding RNAs(lncRNAs)participate in tumor progression by regulating the expression of vascular endothelial growth factor in metastatic CRC.Thus,targeting specific lncRNAs may provide some new hope for anti-angiogenic strategies.Through analyzing data from both clinical samples and The Cancer Genome Atlas database,we found that the lncRNA LINC01503 was specifically upregulated in CRC tissues and was associated with tumor progression and poor overall survival.We also demonstrated that LINC01503 enhanced the capacity for tube formation and migration of vascular endothelial cells,thus promoting CRC tumorigenesis by upregulating vascular endothelial growth factor A(VEGFA)expression in CRC cells.Mechanistically,LINC01503 promoted the expression of VEGFA by simultaneously regulating both mRNA and protein stability of VEGFA by binding to miR-342-3p and the chaperone HSP60,respectively.The upregulation of LINC01503 in CRC cells was attributed to the CREB-binding protein CBP/p300-mediated H3K27 acetylation of the LINC01503 promoter region.Taken together,our findings clarify the mechanism by which LINC01503 may promote CRC angiogenesis,implying that LINC01503 may serve as a potential prognostic biomarker and therapeutic target for CRC.
基金Supported by Guangzhou Basic and Applied Basic Research Foundation,No.202201010121Medical Joint Fund of Jinan University,No.YXZY2024014 and No.YXJC2022001+2 种基金Hospital Achievement Transformation and Cultivation Project,No.ZH201911the Key Discipline Project of Guangdong Province,No.2019-GDXK-0016and the Medical Science and Technology Research Foundation of Guangdong Province,No.B2021145.
文摘BACKGROUND Cervical cancer(CC)stem cell-like cells(CCSLCs),defined by the capacity of differentiation and self-renewal and proliferation,play a significant role in the progression of CC.However,the molecular mechanisms regulating their self-renewal are poorly understood.Therefore,elucidation of the epigenetic mechanisms that drive cancer stem cell self-renewal will enhance our ability to improve the effectiveness of targeted therapies for cancer stem cells.AIM To explore how DNA methyltransferase 1(DNMT1)/miR-342-3p/Forkhead box M1(FoxM1),which have been shown to have abnormal expression in CCSLCs,and their signaling pathways could stimulate self-renewal-related stemness in CCSLCs.METHODS Sphere-forming cells derived from CC cell lines HeLa,SiHa and CaSki served as CCSLCs.Self-renewal-related stemness was identified by determining sphere and colony formation efficiency,CD133 and CD49f protein level,and SRY-box transcription factor 2 and octamer-binding transcription factor 4 mRNA level.The microRNA expression profiles between HeLa cells and HeLa-derived CCSLCs or mRNA expression profiles that HeLaderived CCSLCs were transfected with or without miR-342-3p mimic were compared using quantitative PCR analysis.The expression levels of DNMT1 mRNA,miR-342-3p,and FoxM1 protein were examined by quantitative real-time PCR and western blotting.In vivo carcinogenicity was assessed using a mouse xenograft model.The functional effects of the DNMT1/miR-342-3p/FoxM1 axis were examined by in vivo and in vitro gain-of-activity and loss-of-activity assessments.Interplay among DNMT1,miR-342-3p,and FoxM1 was tested by methylationspecific PCR and a respective luciferase reporter assay.RESULTS CCSLCs derived from the established HeLa cell lines displayed higher self-renewal-related stemness,including enhanced sphere and colony formation efficiency,increased CD133 and CD49f protein level,and heightened transcriptional quantity of stemness-related factors SRY-box transcription factor 2 and octamer-binding transcription factor 4 in vitro as well as a stronger tumorigenic potential in vivo compared to their parental cells.Moreover,quantitative PCR showed that the miR-342-3p level was downregulated in HeLa-derived CCSLCs compared to HeLa cells.Its mimic significantly decreased DNMT1 and FoxM1 mRNA expression levels in CCSLCs.Knockdown of DNMT1 or miR-342-3p mimic transfection suppressed DNMT1 expression,increased miR-342-3p quantity by promoter demethylation,and inhibited CCSLC self-renewal.Inhibition of FoxM1 by shRNA transfection also resulted in the attenuation of CCSLC self-renewal but had little effect on the DNMT1 activity and miR-342-3p expression.Furthermore,the loss of CCSLC self-renewal exerted by miR-342-3p mimic was inverted by the overexpression of DNMT1 or FoxM1.Furthermore,DNMT1 and FoxM1 were recognized as straight targets by miR-342-3p in HeLa-derived CCSLCs.CONCLUSION Our findings suggested that a novel DNMT1/miR-342-3p/FoxM1 signal axis promotes CCSLC self-renewal and presented a potential target for the treatment of CC through suppression of CCSLC self-renewal.However,this pathway has been previously implicated in CC,as evidenced by prior studies showing miR-342-3p-mediated downregulation of FoxM1 in cervical cancer cells.Additionally,research on liver cancer further supports the involvement of miR-342-3p in suppressing FoxM1 expression.While our study contributed to this body of knowledge,we did not present a completely novel axis but reinforced the therapeutic potential of targeting the DNMT1/miR-342-3p/FoxM1 axis to suppress CCSLC self-renewal in CC treatment.
文摘目的:探讨miR-342-3p在鼻咽癌(NPC)中的表达及其意义。方法:miRNA芯片检测人永生化鼻咽上皮细胞系NP69和低分化NPC细胞CNE2中miRNA的差异表达;实时荧光定量PCR(qPCR)法检测NPC组织中miR-342-3p的表达;GO and Pathway分析miR-342-3p在NPC发生发展中可能涉及的功能及其参与的信号通路。结果:miRNA芯片结果表明,miR-342-3p在NPC细胞CNE2中低表达,qPCR结果进一步证实miR-342-3p在NPC组织中低表达;GO分析表明,具有显著差异的10个功能中,与NPC发生发展关系较为密切的是miR-342-3p。Pathway分析显示,miR-342-3p与丝裂原应激活化蛋白(MAPK)信号通路相关性较大。结论:miR-342-3p的低表达可能与NPC的发生和发展相关。
