Background:Pancreatic adenocarcinoma(PAAD)represents a highly fatal form of cancer.The 5-year survival rate for patients with this disease is only around 10%.A significant hurdle in its management is the absence of ch...Background:Pancreatic adenocarcinoma(PAAD)represents a highly fatal form of cancer.The 5-year survival rate for patients with this disease is only around 10%.A significant hurdle in its management is the absence of characteristic early-stage symptoms.As a result,a large majority of pancreatic cancer patients are diagnosed when the disease has reached an advanced stage or has metastasized.Consequently,taking measures to suppress the occurrence of metastasis in pancreatic cancer can bring about a substantial improvement in patients'survival rates and overall prognosis.SKIL,known to promote cancer progression,is implicated in cell proliferation,epithelial–mesenchymal transition(EMT),and metastasis,but its specific function in pancreatic cancer remains unclear.Methods:We investigated the effects of SKIL on the proliferation,apoptosis,and metastasis of pancreatic cancer cells.Through ChIP-seq,we identified the SKIL downstream target gene and further explored the mechanism by which SKIL regulates the metastasis of pancreatic cancer cells through functional experiments and Western blot.Results:A high level of SKIL expression is associated with an unfavorable prognosis in PAAD;it promotes cell migration and EMT.Through ChIP-seq analysis,we identified that SKIL inhibits TSPYL2,a nuclear protein regulating the TGF-βpathway by binding to the TGFB1 promoter.Further studies carried out by us confirmed that SKIL modulates the TGF-βpathway via TSPYL2,facilitating EMT and metastasis in pancreatic cancer cells,independent of Smad4.Conclusions:These findings reveal a novel regulatory mechanism involving SKIL,TSPYL2,and the TGF-βpathway,offering new therapeutic targets for PAAD.展开更多
Cullin-RING E3 ubiquitin ligases(CRLs),the largest family of multi-subunit E3 ubiquitin ligases in eukaryotic cells,represent core cellular machinery for executing protein degradation and maintaining proteostasis.Here...Cullin-RING E3 ubiquitin ligases(CRLs),the largest family of multi-subunit E3 ubiquitin ligases in eukaryotic cells,represent core cellular machinery for executing protein degradation and maintaining proteostasis.Here,we asked what roles Cullin proteins play in human mesenchymal stem cell(hMSC)homeostasis and senescence.To this end,we conducted a comparative aging phenotype analysis by individually knocking down Cullin members in three senescence models:replicative senescent hMSCs,Hutchinson-Gilford Progeria Syndrome hMSCs,and Werner syndrome hMSCs.Among all family members,we found that CUL2 deficiency rendered hMSCs the most susceptible to senescence.To investigate CUL2-specific underlying mechanisms,we then applied CRISPR/Cas9-mediated gene editing technology to generate CUL2-deficient human embryonic stem cells(hESCs).When we differentiated these into h MSCs,we found that CUL2 deletion markedly accelerates hMSC senescence.Importantly,we identified that CUL2 targets and promotes ubiquitin proteasome-mediated degradation of TSPYL2(a known negative regulator of proliferation)through the substrate receptor protein APPBP2,which in turn downregulates one of the canonical aging marker-P21^(waf1/cip1),and thereby delays senescence.Our work provides important insights into how CRL2^(APPBP2)-mediated TSPYL2 degradation counteracts hMSC senescence,providing a molecular basis for directing intervention strategies against aging and aging-related diseases.展开更多
基金supported by the National Natural Science Foundation Fund(Grant Nos.82372696,82172988)the Chinese Academy of Medical Sciences(CAMS)Innovation Fund for Medical Sciences(CIFMS)(Grant No.2021‐I2M‐1‐014)the Cooperation Fund of CHCAMS and SZCH(Grant No.CFA202201006).
文摘Background:Pancreatic adenocarcinoma(PAAD)represents a highly fatal form of cancer.The 5-year survival rate for patients with this disease is only around 10%.A significant hurdle in its management is the absence of characteristic early-stage symptoms.As a result,a large majority of pancreatic cancer patients are diagnosed when the disease has reached an advanced stage or has metastasized.Consequently,taking measures to suppress the occurrence of metastasis in pancreatic cancer can bring about a substantial improvement in patients'survival rates and overall prognosis.SKIL,known to promote cancer progression,is implicated in cell proliferation,epithelial–mesenchymal transition(EMT),and metastasis,but its specific function in pancreatic cancer remains unclear.Methods:We investigated the effects of SKIL on the proliferation,apoptosis,and metastasis of pancreatic cancer cells.Through ChIP-seq,we identified the SKIL downstream target gene and further explored the mechanism by which SKIL regulates the metastasis of pancreatic cancer cells through functional experiments and Western blot.Results:A high level of SKIL expression is associated with an unfavorable prognosis in PAAD;it promotes cell migration and EMT.Through ChIP-seq analysis,we identified that SKIL inhibits TSPYL2,a nuclear protein regulating the TGF-βpathway by binding to the TGFB1 promoter.Further studies carried out by us confirmed that SKIL modulates the TGF-βpathway via TSPYL2,facilitating EMT and metastasis in pancreatic cancer cells,independent of Smad4.Conclusions:These findings reveal a novel regulatory mechanism involving SKIL,TSPYL2,and the TGF-βpathway,offering new therapeutic targets for PAAD.
基金supported by the National Key Research and Development Program of China(2020YFA0804000,2022YFA1103700,2020YFA0112200,2021YFF1201000,the STI2030-Major Projects-2021ZD0202400,2022YFA1103800)the National Natural Science Foundation of China(82201714,81921006,82125011,92149301,92168201,91949209,92049304,92049116,32121001,82192863,82122024,82071588,32000500,82271600,82001477,82201727)+12 种基金the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA16000000)CAS Project for Young Scientists in Basic Research(YSBR-076,YSBR-012)the Program of the Beijing Natural Science Foundation(Z190019)the Fellowship of China Postdoctoral Science Foundation(2022M712216)the Project for Technology Development of Beijing-affiliated Medical Research Institutes(11000023T000002036310)the Pilot Project for Public Welfare Development and Reform of Beijing-affiliated Medical Research Institutes(11000022T000000461062)Youth Innovation Promotion Association of CAS(E1CAZW0401,2022083,2023092)Young Elite Scientists Sponsorship Program by CAST(YESS20200012,YESS20210002)the Informatization Plan of Chinese Academy of Sciences(CAS-WX2021SF-0301,CAS-WX2022SDC-XK14,CASWX2021SF-0101)New Cornerstone Science Foundation through the XPLORER PRIZE(2021-1045)Excellent Young Talents Program of Capital Medical University(12300927)Excellent Young Talents Training Program for the Construction of Beijing Municipal University Teacher Team(BPHR202203105)Beijing Hospitals Authority Youth Programme(QML20230806)。
文摘Cullin-RING E3 ubiquitin ligases(CRLs),the largest family of multi-subunit E3 ubiquitin ligases in eukaryotic cells,represent core cellular machinery for executing protein degradation and maintaining proteostasis.Here,we asked what roles Cullin proteins play in human mesenchymal stem cell(hMSC)homeostasis and senescence.To this end,we conducted a comparative aging phenotype analysis by individually knocking down Cullin members in three senescence models:replicative senescent hMSCs,Hutchinson-Gilford Progeria Syndrome hMSCs,and Werner syndrome hMSCs.Among all family members,we found that CUL2 deficiency rendered hMSCs the most susceptible to senescence.To investigate CUL2-specific underlying mechanisms,we then applied CRISPR/Cas9-mediated gene editing technology to generate CUL2-deficient human embryonic stem cells(hESCs).When we differentiated these into h MSCs,we found that CUL2 deletion markedly accelerates hMSC senescence.Importantly,we identified that CUL2 targets and promotes ubiquitin proteasome-mediated degradation of TSPYL2(a known negative regulator of proliferation)through the substrate receptor protein APPBP2,which in turn downregulates one of the canonical aging marker-P21^(waf1/cip1),and thereby delays senescence.Our work provides important insights into how CRL2^(APPBP2)-mediated TSPYL2 degradation counteracts hMSC senescence,providing a molecular basis for directing intervention strategies against aging and aging-related diseases.
