The tumor microenvironment is a complex network of cells,extracellular matrix,and signaling molecules that plays a critical role in tumor progression and metastasis.Lymphatic and blood vessels are major routes for sol...The tumor microenvironment is a complex network of cells,extracellular matrix,and signaling molecules that plays a critical role in tumor progression and metastasis.Lymphatic and blood vessels are major routes for solid tumor metastasis and essential parts of tumor drainage conduits.However,recent studies have shown that lymphatic endothelial cells(LECs)and blood endothelial cells(BECs)also play multifaceted roles in the tumor microenvironment beyond their structural functions,particularly in hepatocellular carcinoma(HCC).This comprehensive review summarizes the diverse roles played by LECs and BECs in HCC,including their involvement in angiogenesis,immune modulation,lymphangiogenesis,and metastasis.By providing a detailed account of the complex interplay between LECs,BECs,and tumor cells,this review aims to shed light on future research directions regarding the immune regulatory function of LECs and potential therapeutic targets for HCC.展开更多
Objective:Lymphatic endothelial cell(LEC)proliferation is essential for lymphangiogenesis.Hypoxia induces lymphangiogenesis,but it directly inhibits LEC proliferation and the underlying mechanisms have not been fully ...Objective:Lymphatic endothelial cell(LEC)proliferation is essential for lymphangiogenesis.Hypoxia induces lymphangiogenesis,but it directly inhibits LEC proliferation and the underlying mechanisms have not been fully understood.The aim of this study was to investigate the role of carcinoembryonic antigen-related cell adhesion molecule 1(CEACAM1)in hypoxia-repressed LEC proliferation.Methods:Human dermal lymphatic endothelial cells(HDLECs)were cultured under normoxic or hypoxic conditions,and cell proliferation was determined using MTT or CCK-8 assays.CEACAM1 expression was silenced by siRNA transfection.Activation of mitogen-activated protein kinases(MAPKs)was examined by Western blotting and blocked by specific inhibitors.Results:Under hypoxia,HDLECs proliferation was suppressed and CEACAM1 expression was downregulated.Silence of CEACAM1 in normoxia inhibited HDLECs proliferation and did not further decrease proliferation in HDLECs in response to hypoxia,suggesting that CEACAM1 may mediate hypoxia-induced inhibition of HDLECs proliferation.In addition,silence of CEACAM1 increased phosphorylation of MAPK molecules:extracellular signal-regulated kinase(ERK),p38 MAPK and Jun N-terminal kinase(JNK)in HDLECs.However,only inhibition of the JNK pathway rescued the reduction of HDLEC proliferation induced by CEACAM1 silence.Conclusion:Our results suggested that hypoxia downregulates CEACAM1 expression by activation of the JNK pathway,leading to inhibition of HDLEC proliferation.These findings may help to understand the mechanisms of LEC-specific response to hypoxia and develop novel therapies for pathological lymphangiogenesis.展开更多
Lymphatic metastasis is a continuous and complicated process. The detailed mechanisms of lymphatic metastasis are still not very clear, despite considerable research efforts in recent years. Previously, it was commonl...Lymphatic metastasis is a continuous and complicated process. The detailed mechanisms of lymphatic metastasis are still not very clear, despite considerable research efforts in recent years. Previously, it was commonly accepted that there were no lymphatic vessels in the primary tumor. However, recent studies have demonstrated that lymphatic vessels are detectable in certain types of cancer, and more and more evidence has shown that cancer cells invade into local lymph nodes mainly via peritumoral lymphatic vessels, Moreover, activated endothelial cells may also be important, having an influence on lymphatic metastasis of cancer cells. This article, based on recent research findings, provides an in-depth discussion of the relationship between lymphangiogenesis, tumor-derived lymphatic endothelial cells and lymphatic metastasis in head and neck cancer.展开更多
Objective:To study explores the effect of HLEC on the secreted proteins of epithelial ovarian cancer(EOC)cells(SKOV3-PM4)with directional highly lymphatic metastasis.Methods:Supernatants of four groups of cultured cel...Objective:To study explores the effect of HLEC on the secreted proteins of epithelial ovarian cancer(EOC)cells(SKOV3-PM4)with directional highly lymphatic metastasis.Methods:Supernatants of four groups of cultured cells,namely,SKOV3(A),SKOV3+HLEC(B),SKOV3-PM4(C),SKOV3-PM4+HLEC(D),were collected,and their proteins were detected by antibody arrays and iTRAQ-2D-LC-MALDITOF/TOF/MS.Significantly differential proteins were further analyzed via bioinformatics and validated in human serums and cell media via ELISA.Results:Results of antibody arrays and mass spectrometry demonstrated that GRN and VEGFA were upregulated in group C(compared with group A),whereas IGFBP7 and SPARC were downregulated in group D(compared with group C).Comprehensive bioinformatics analysis results showed that IGFBP7 and VEGFA were closely linked to each other.Further validation with serums showed statistical significance in VEGFA and IGFBP7 levels among groups of patients with ovarian cancers,benign tumors,and control groups.Two proteins were upegulated in the first group.VEGFA in the control group was downregulated.For IGFBP,upregulation in the control group and down-regulation in the first group were also observed.Conclusion:The HLEC microenvironment is closely associated with directional metastasis to lymph nodes and with differential proteins including cell stromal proteins and adhesion factors.The upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 are closely associated with directional metastasis to lymph nodes in EOC cells.展开更多
OBJECTIVE To examine the expression of vascular endothelial growth factor C(VEGF-C)in human esophageal squamous cell carcinoma(ESCC), and to clarify its role in lymphatic metastasis in ESCC patients. METHODS Esophagea...OBJECTIVE To examine the expression of vascular endothelial growth factor C(VEGF-C)in human esophageal squamous cell carcinoma(ESCC), and to clarify its role in lymphatic metastasis in ESCC patients. METHODS Esophageal carcinoma EC9706 cel s and samples from 49 patients with primary ESCC were investigated by using S-P immunohisto- chemistry(IHC),the semi-quantitative reverse transcriptase-polymerase chain reaction(RT-PCR)and in situ hybridization(ISH)methods for VEGF-C expression. RESULTS VEGF-C positive expression was found in EC9706 cells through IHC,ISH and RT-PCR.Positive IHC for VEGF-C was observed in 36 of 49 cases of ESCC.There was a significant difference between the expres- sion of VEGF-C in a lymph-node-positive group compared to a node-nega- tive group(χ2=4.7,P<0.05).Positive ISH for VEGF-C mRNA was observed in 23 of 49 cases of ESCC.There was a significant difference between the expression of VEGF-C in the lymph-node-positive group and node-negative group(χ2=31.3,P<0.01).The expression of VEGF-C was significantly higher in the lymph-node-positive group compared to the node-negative group.Of 49 ESCC tissues,RT-PCR for VEGF-C mRNA was observed positively in 29 cases.There was a significant difference between the expression of VEGF-C in the lymph-node-positive group and node-negative group(χ2=23.3, P<0.01).The expression of VEGF-C was significantly higher in the lymph- node-positive group compared to the node-negative group.Expressions of VEGF-C were not significantly associated with age,gender,and pathological grade.There was a relationship between VEGF-C mRNA expressions by RT-PCR and ISH(χ2=18.5,P<0.01)in ESCC cases,but with no significant difference between the two methods. CONCLUSION VEGF-C expression may induce lymphangiogenesis in human ESCC.There was a close correlation between VEGF-C expression and lymph node metastasis.VEGF-C can serve as a useful prognostic factor for ESCC patients.展开更多
Background Pancreatic cancer is one of the most aggressive human malignancies. Lymphangiogenesis plays an important role in lymph node metastasis of many solid tumors. It is well known that low molecular weight hepari...Background Pancreatic cancer is one of the most aggressive human malignancies. Lymphangiogenesis plays an important role in lymph node metastasis of many solid tumors. It is well known that low molecular weight heparins （LMWHs） can inhibit cell growth, cell invasion and angiogenesis, which are key processes in tumor progression. Methods We measured the expression of vascular endothelial growth factor C （VEGF-C） in pancreatic cancer cells （PANC-1） using reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting. We used an in vitro assay to evaluate the anti-lymphangiogenic effect of an LMWH, Fragmin, on human lymphatic endothelial cell （HLEC） proliferation. Results Fragmin at a low concentration can effectively inhibits HLEC proliferation induced by VEGF-C. VEGF-C secreted by PANC-1 cells stimulated HLEC proliferation. Low concentration LMWH suppressed HLEC proliferation induced by VEGF-C but did not affect proliferation or VEGF-C expression of PANC-1 cells, whereas high concentrations of LMWH inhibited PANC-1 cell proliferation. Conclusions These results suggest that VEGF-C released by cancer cells plays an important role in promoting HLEC proliferation. The LMWH Fragmin has anti-lymphangiogenic effects and may inhibit lymphatic metastasis in pancreatic cancer.展开更多
Lymphangiogenesis, the growth of new lymphatic vessels, has long been regarded as a putative efficient pathway to neoplastic metastization. Recent results have shown the necessity of lymphatic molecular markers and gr...Lymphangiogenesis, the growth of new lymphatic vessels, has long been regarded as a putative efficient pathway to neoplastic metastization. Recent results have shown the necessity of lymphatic molecular markers and growth factors for lymphangiogenesis. Importantly, lymphatic endothelial receptor tyrosine kinase VEGFR-3 and its ligands VEGF-C and VEGF-D play crucial roles in promoting lymphatic vascular growth both during development and in pathological conditions. Isolation of pure cultures of lymphatic and blood vascular endothelial cells and systematic characterization of their transcriptomes provide useful cell culture models and novel potential vascular markers and offer further insights into the lymphatic vascular biology. Ectopic expression of the lymphatic endothelial specific homeobox transcription factor Prox1 in blood endothelial cells results in a shift in the gene expression profile towards the lymphatic endothelial phenotype. It demonstrates the plasticity of endothelial cells and offers the possibility of transcriptional reprogramming of vascular endothelial cells as future putative therapeutic applications.展开更多
Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolyti...Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.展开更多
基金Supported by National Natural Science Foundation of China,No.81702923,and No.81971503Open Project of State Key Laboratory of Medical Immunology,No.NKLMI2023K03+1 种基金Shanghai Shen Kang Hospital Development Center Clinical Science and Technology Innovation Project,No.SHDC12020104Basic Medical Research Project of Naval Medical University,No.2022QN072.
文摘The tumor microenvironment is a complex network of cells,extracellular matrix,and signaling molecules that plays a critical role in tumor progression and metastasis.Lymphatic and blood vessels are major routes for solid tumor metastasis and essential parts of tumor drainage conduits.However,recent studies have shown that lymphatic endothelial cells(LECs)and blood endothelial cells(BECs)also play multifaceted roles in the tumor microenvironment beyond their structural functions,particularly in hepatocellular carcinoma(HCC).This comprehensive review summarizes the diverse roles played by LECs and BECs in HCC,including their involvement in angiogenesis,immune modulation,lymphangiogenesis,and metastasis.By providing a detailed account of the complex interplay between LECs,BECs,and tumor cells,this review aims to shed light on future research directions regarding the immune regulatory function of LECs and potential therapeutic targets for HCC.
基金supported by grants from the National Natural Science Foundation of China(No.81873473 and No.91939110)Academic Promotion Program of Shandong First Medical University(No.2019QL014)Shandong Taishan Scholarship(Ju Liu).
文摘Objective:Lymphatic endothelial cell(LEC)proliferation is essential for lymphangiogenesis.Hypoxia induces lymphangiogenesis,but it directly inhibits LEC proliferation and the underlying mechanisms have not been fully understood.The aim of this study was to investigate the role of carcinoembryonic antigen-related cell adhesion molecule 1(CEACAM1)in hypoxia-repressed LEC proliferation.Methods:Human dermal lymphatic endothelial cells(HDLECs)were cultured under normoxic or hypoxic conditions,and cell proliferation was determined using MTT or CCK-8 assays.CEACAM1 expression was silenced by siRNA transfection.Activation of mitogen-activated protein kinases(MAPKs)was examined by Western blotting and blocked by specific inhibitors.Results:Under hypoxia,HDLECs proliferation was suppressed and CEACAM1 expression was downregulated.Silence of CEACAM1 in normoxia inhibited HDLECs proliferation and did not further decrease proliferation in HDLECs in response to hypoxia,suggesting that CEACAM1 may mediate hypoxia-induced inhibition of HDLECs proliferation.In addition,silence of CEACAM1 increased phosphorylation of MAPK molecules:extracellular signal-regulated kinase(ERK),p38 MAPK and Jun N-terminal kinase(JNK)in HDLECs.However,only inhibition of the JNK pathway rescued the reduction of HDLEC proliferation induced by CEACAM1 silence.Conclusion:Our results suggested that hypoxia downregulates CEACAM1 expression by activation of the JNK pathway,leading to inhibition of HDLEC proliferation.These findings may help to understand the mechanisms of LEC-specific response to hypoxia and develop novel therapies for pathological lymphangiogenesis.
文摘Lymphatic metastasis is a continuous and complicated process. The detailed mechanisms of lymphatic metastasis are still not very clear, despite considerable research efforts in recent years. Previously, it was commonly accepted that there were no lymphatic vessels in the primary tumor. However, recent studies have demonstrated that lymphatic vessels are detectable in certain types of cancer, and more and more evidence has shown that cancer cells invade into local lymph nodes mainly via peritumoral lymphatic vessels, Moreover, activated endothelial cells may also be important, having an influence on lymphatic metastasis of cancer cells. This article, based on recent research findings, provides an in-depth discussion of the relationship between lymphangiogenesis, tumor-derived lymphatic endothelial cells and lymphatic metastasis in head and neck cancer.
基金supported by the National Natural Science Foundation of China (Grant No. 81060218)Guangxi Natural Science Foundation (Grant No. 2012GXNSFAA053157)
文摘Objective:To study explores the effect of HLEC on the secreted proteins of epithelial ovarian cancer(EOC)cells(SKOV3-PM4)with directional highly lymphatic metastasis.Methods:Supernatants of four groups of cultured cells,namely,SKOV3(A),SKOV3+HLEC(B),SKOV3-PM4(C),SKOV3-PM4+HLEC(D),were collected,and their proteins were detected by antibody arrays and iTRAQ-2D-LC-MALDITOF/TOF/MS.Significantly differential proteins were further analyzed via bioinformatics and validated in human serums and cell media via ELISA.Results:Results of antibody arrays and mass spectrometry demonstrated that GRN and VEGFA were upregulated in group C(compared with group A),whereas IGFBP7 and SPARC were downregulated in group D(compared with group C).Comprehensive bioinformatics analysis results showed that IGFBP7 and VEGFA were closely linked to each other.Further validation with serums showed statistical significance in VEGFA and IGFBP7 levels among groups of patients with ovarian cancers,benign tumors,and control groups.Two proteins were upegulated in the first group.VEGFA in the control group was downregulated.For IGFBP,upregulation in the control group and down-regulation in the first group were also observed.Conclusion:The HLEC microenvironment is closely associated with directional metastasis to lymph nodes and with differential proteins including cell stromal proteins and adhesion factors.The upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 are closely associated with directional metastasis to lymph nodes in EOC cells.
基金This work was supported by a grant from theNational Natural Science Foundation of China(No.30470779)the Henan InnovationProject for University Prominent ResearchTalents(No.2006KYCX016)
文摘OBJECTIVE To examine the expression of vascular endothelial growth factor C(VEGF-C)in human esophageal squamous cell carcinoma(ESCC), and to clarify its role in lymphatic metastasis in ESCC patients. METHODS Esophageal carcinoma EC9706 cel s and samples from 49 patients with primary ESCC were investigated by using S-P immunohisto- chemistry(IHC),the semi-quantitative reverse transcriptase-polymerase chain reaction(RT-PCR)and in situ hybridization(ISH)methods for VEGF-C expression. RESULTS VEGF-C positive expression was found in EC9706 cells through IHC,ISH and RT-PCR.Positive IHC for VEGF-C was observed in 36 of 49 cases of ESCC.There was a significant difference between the expres- sion of VEGF-C in a lymph-node-positive group compared to a node-nega- tive group(χ2=4.7,P<0.05).Positive ISH for VEGF-C mRNA was observed in 23 of 49 cases of ESCC.There was a significant difference between the expression of VEGF-C in the lymph-node-positive group and node-negative group(χ2=31.3,P<0.01).The expression of VEGF-C was significantly higher in the lymph-node-positive group compared to the node-negative group.Of 49 ESCC tissues,RT-PCR for VEGF-C mRNA was observed positively in 29 cases.There was a significant difference between the expression of VEGF-C in the lymph-node-positive group and node-negative group(χ2=23.3, P<0.01).The expression of VEGF-C was significantly higher in the lymph- node-positive group compared to the node-negative group.Expressions of VEGF-C were not significantly associated with age,gender,and pathological grade.There was a relationship between VEGF-C mRNA expressions by RT-PCR and ISH(χ2=18.5,P<0.01)in ESCC cases,but with no significant difference between the two methods. CONCLUSION VEGF-C expression may induce lymphangiogenesis in human ESCC.There was a close correlation between VEGF-C expression and lymph node metastasis.VEGF-C can serve as a useful prognostic factor for ESCC patients.
文摘Background Pancreatic cancer is one of the most aggressive human malignancies. Lymphangiogenesis plays an important role in lymph node metastasis of many solid tumors. It is well known that low molecular weight heparins （LMWHs） can inhibit cell growth, cell invasion and angiogenesis, which are key processes in tumor progression. Methods We measured the expression of vascular endothelial growth factor C （VEGF-C） in pancreatic cancer cells （PANC-1） using reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting. We used an in vitro assay to evaluate the anti-lymphangiogenic effect of an LMWH, Fragmin, on human lymphatic endothelial cell （HLEC） proliferation. Results Fragmin at a low concentration can effectively inhibits HLEC proliferation induced by VEGF-C. VEGF-C secreted by PANC-1 cells stimulated HLEC proliferation. Low concentration LMWH suppressed HLEC proliferation induced by VEGF-C but did not affect proliferation or VEGF-C expression of PANC-1 cells, whereas high concentrations of LMWH inhibited PANC-1 cell proliferation. Conclusions These results suggest that VEGF-C released by cancer cells plays an important role in promoting HLEC proliferation. The LMWH Fragmin has anti-lymphangiogenic effects and may inhibit lymphatic metastasis in pancreatic cancer.
文摘Lymphangiogenesis, the growth of new lymphatic vessels, has long been regarded as a putative efficient pathway to neoplastic metastization. Recent results have shown the necessity of lymphatic molecular markers and growth factors for lymphangiogenesis. Importantly, lymphatic endothelial receptor tyrosine kinase VEGFR-3 and its ligands VEGF-C and VEGF-D play crucial roles in promoting lymphatic vascular growth both during development and in pathological conditions. Isolation of pure cultures of lymphatic and blood vascular endothelial cells and systematic characterization of their transcriptomes provide useful cell culture models and novel potential vascular markers and offer further insights into the lymphatic vascular biology. Ectopic expression of the lymphatic endothelial specific homeobox transcription factor Prox1 in blood endothelial cells results in a shift in the gene expression profile towards the lymphatic endothelial phenotype. It demonstrates the plasticity of endothelial cells and offers the possibility of transcriptional reprogramming of vascular endothelial cells as future putative therapeutic applications.
基金supported by the National Natural Science Foundation of China(No.81974321)the Discipline Innovation and Talent Introduction Program to Universities from Ministry of Education of China(Project 111)+3 种基金the Tackling Project for Science and Technology of Xinxiang City(No.GG2019003)the Natural Science Foundation of Henan Province of China(No.212300410173)the State Administration of Traditional Chinese Medicine Young Qi Huang Scholar,and the Innovation Team Project of Scientific Research of Traditional Chinese Medicine of Shanghai Health Committee(No.2022CX001)China.
文摘Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.
