Objective To discover critical genes contributing to the sternness and maintenance of spermatogonial stem cells(SSCs)and provide new insights into the function of the leucine-rich repeat(LRR)family member Lrrc34(leudn...Objective To discover critical genes contributing to the sternness and maintenance of spermatogonial stem cells(SSCs)and provide new insights into the function of the leucine-rich repeat(LRR)family member Lrrc34(leudne-rich repeat-containing 34)in SSCs from mice.Methods Bioinformatic methods,including differentially expressed gene(DEG),gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses,were used to uncover latent pluripotency-related genes.Reverse transcription-polymerase chain reaction(RT-PCR)and immunofluorescence analyses were utilized to verify the mRNA and protein expression levels,respectively.RNA interference of Lrrc34 using siRNA was performed to detect its transient impact on SSCs.Results Eight DEGs between ID4-EGFP+(G)and ID4-EGFP+/TSPAN8Hig11(TH),eight DEGs between G and ID4-EGFP+/TSPAN8Uw(TL)and eleven DEGs between TH and TL were discovered,and eleven proteinprotein interaction(PPI)modules were found to be significant in the PPI network of DEGs.One of the DEGs,Lrrc34,was selected as a potential pluripotency?related gene due to its differential expression among ID4-EGFP+spermatogonia subsets and its interaction with fibroblast growth factor 2 in the fifth module.Immunofluorescence experiments exhibited specific expression of Lrrc34 in a subpopulation of undifferentiated spermatogonia marked by LIN28A,and RT-PCR experiments confirmed the high expression of Lrrc34 in SSCs from P7 and adult mice.The transient knockdown of Lrrc34 in SSCs resulted in reduced colony sizes and significant changes in the transcriptome and apoptotic pathways.Conclusion Lrrc34 is highly expressed in mouse SSCs and is required for SSC proliferation in vitro through effects on transcriptome and signaling transduction pathways.展开更多
Prolonged activation of nuclear factor(NF)-кB signaling significantly contributes to the development of colorectal cancer(CRC).New therapeutic opportunities are emerging from targeting this distorted cell signaling t...Prolonged activation of nuclear factor(NF)-кB signaling significantly contributes to the development of colorectal cancer(CRC).New therapeutic opportunities are emerging from targeting this distorted cell signaling transduction.Here,we discovered the critical role of RING finger 138(RNF138)in CRC tumorigenesis through regulating the NF-кB signaling,which is independent of its Ubiquitin-E3 ligase activity involved in DNA damage response.RNF138^(−/−) mice were hyper-susceptible to the switch from colitis to aggressive malignancy,which coincided with sustained aberrant NF-кB signaling in the colonic cells.Furthermore,RNF138 suppresses the activation of NF-кB signaling pathway through preventing the translocation of NIK and IKK-Beta Binding Protein(NIBP)to the cytoplasm,which requires the ubiquitin interaction motif(UIM)domain.More importantly,we uncovered a significant correlation between poor prognosis and the downregulation of RNF138 associated with reinforced NF-кB signaling in clinical settings,raising the possibility of RNF138 dysregulation as an indicator for the therapeutic intervention targeting NF-кB signaling.Using the xenograft models built upon either RNF138-dificient CRC cells or the cells derived from the RNF138-dysregulated CRC patients,we demonstrated that the inhibition of NF-кB signaling effectively hampered tumor growth.Overall,our work defined the pathogenic role of aberrant NF-кB signaling due to RNF138 downregulation in the cascade events from the colitis switch to colonic neoplastic transformation and progression,and also highlights the possibility of targeting the NF-кB signaling in treating specific subtypes of CRC indicated by RNF138-ablation.展开更多
基金grants from the National Key Research and Development Program of China(2018YFC1003500)the CAMS Innovation Fund for Medical Sciences(2017-I2M-3-009,2016-12M-1-001,and 2017-I2M-1-004)+1 种基金the National Natural Science Foundation of China(81672472 and 31970794)the State Key Laboratory Special Fund from the Ministry of Science of China(2060204).
文摘Objective To discover critical genes contributing to the sternness and maintenance of spermatogonial stem cells(SSCs)and provide new insights into the function of the leucine-rich repeat(LRR)family member Lrrc34(leudne-rich repeat-containing 34)in SSCs from mice.Methods Bioinformatic methods,including differentially expressed gene(DEG),gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses,were used to uncover latent pluripotency-related genes.Reverse transcription-polymerase chain reaction(RT-PCR)and immunofluorescence analyses were utilized to verify the mRNA and protein expression levels,respectively.RNA interference of Lrrc34 using siRNA was performed to detect its transient impact on SSCs.Results Eight DEGs between ID4-EGFP+(G)and ID4-EGFP+/TSPAN8Hig11(TH),eight DEGs between G and ID4-EGFP+/TSPAN8Uw(TL)and eleven DEGs between TH and TL were discovered,and eleven proteinprotein interaction(PPI)modules were found to be significant in the PPI network of DEGs.One of the DEGs,Lrrc34,was selected as a potential pluripotency?related gene due to its differential expression among ID4-EGFP+spermatogonia subsets and its interaction with fibroblast growth factor 2 in the fifth module.Immunofluorescence experiments exhibited specific expression of Lrrc34 in a subpopulation of undifferentiated spermatogonia marked by LIN28A,and RT-PCR experiments confirmed the high expression of Lrrc34 in SSCs from P7 and adult mice.The transient knockdown of Lrrc34 in SSCs resulted in reduced colony sizes and significant changes in the transcriptome and apoptotic pathways.Conclusion Lrrc34 is highly expressed in mouse SSCs and is required for SSC proliferation in vitro through effects on transcriptome and signaling transduction pathways.
基金supported by CAMS Innovation Fund for Medical Sciences(CIFMS,2021-I2M-1-066,2017-I2M-4-002 to H.Z.,2021-I2M-1-019 to W.S.,2021-1-I2M-014 to C.Z.L.)the National Natural Science Foundation of China(81972311,82141127 and 81672461 to H.Z.,81672472,and 31970794 to W.S.,81570780 to C.Z.L.,32000586 to K.L.)+4 种基金the Nonprofit Central Research Institute Fund of Chinese Academy of Medical Sciences(2019PT310026 to H.Z.)Sanming Project of Medicine in Shenzhen(SZSM202011010 to H.Z.)the National Key Research and Development Program of China(2018YFC1003500 to W.S.)the State Key Laboratory Special fund from the Ministry of Science(2060204 to W.S.)the State Key Project on Infection Diseases of China(2017ZX10201021-007-003 to H.Z.).
文摘Prolonged activation of nuclear factor(NF)-кB signaling significantly contributes to the development of colorectal cancer(CRC).New therapeutic opportunities are emerging from targeting this distorted cell signaling transduction.Here,we discovered the critical role of RING finger 138(RNF138)in CRC tumorigenesis through regulating the NF-кB signaling,which is independent of its Ubiquitin-E3 ligase activity involved in DNA damage response.RNF138^(−/−) mice were hyper-susceptible to the switch from colitis to aggressive malignancy,which coincided with sustained aberrant NF-кB signaling in the colonic cells.Furthermore,RNF138 suppresses the activation of NF-кB signaling pathway through preventing the translocation of NIK and IKK-Beta Binding Protein(NIBP)to the cytoplasm,which requires the ubiquitin interaction motif(UIM)domain.More importantly,we uncovered a significant correlation between poor prognosis and the downregulation of RNF138 associated with reinforced NF-кB signaling in clinical settings,raising the possibility of RNF138 dysregulation as an indicator for the therapeutic intervention targeting NF-кB signaling.Using the xenograft models built upon either RNF138-dificient CRC cells or the cells derived from the RNF138-dysregulated CRC patients,we demonstrated that the inhibition of NF-кB signaling effectively hampered tumor growth.Overall,our work defined the pathogenic role of aberrant NF-кB signaling due to RNF138 downregulation in the cascade events from the colitis switch to colonic neoplastic transformation and progression,and also highlights the possibility of targeting the NF-кB signaling in treating specific subtypes of CRC indicated by RNF138-ablation.