BACKGROUND Tankyrase 2(TNKS2)is a potential candidate molecular target for the prognosis and treatment of non-small cell lung cancer(NSCLC),but its biological functions are unclear.AIM To investigate the biological fu...BACKGROUND Tankyrase 2(TNKS2)is a potential candidate molecular target for the prognosis and treatment of non-small cell lung cancer(NSCLC),but its biological functions are unclear.AIM To investigate the biological functions of TNKS2 in NSCLC.METHODS Using a lentiviral vector,we generated H647 model cells with TNKS2 knockdown by RNA interference and A549 model cells with TNKS2 overexpression by tran-sfection with a TNKS2 overexpressing plasmid.Increased and decreased exp-ression levels of TNKS2 in the two cell lines were verified using real-time reverse transcriptase-polymerase chain reaction and Western blot analyses.Cell apopto-sis,proliferation,and migration were determined using flow cytometry,carbo-xyfluorescein succinimidyl ester staining,and scratch assay,respectively.Im-munofluorescence staining was conducted to examine TNKS2 andβ-catenin ex-pression levels in the two transfected cell lines and the non-transfected cells.RESULTS TNKS2 mRNA and protein expression was significantly higher in the highly malignant NCI-H647 cells,while it remained at a low level in the less malignant A549 cells.Lentivirus-mediated overexpression of TNKS2 in A549 cells resulted in a 3-fold increase in gene expression and a 1.7-fold increase in protein expression(P<0.01).Conversely,shRNA interference targeting TNKS2 Led to an 8-fold decrease in gene expression and a 3-fold decrease in protein expression(P<0.01)in NCI-H647 cells.Furthermore,the cell apoptosis rate was significantly reduced(50%)and cell migration rate was increased(35%)in the TNKS2 overexpression group than in the control group(P<0.05).In contrast,shTNKS2 promoted apoptosis by more than one fold and reduced migration by 60%(P<0.05).Immunofluorescence analysis revealed enhanced nuclear localization ofβ-catenin fluorescence signal associated with high TNKS2 expression levels.Western blot analysis investigating TNKS2/β-catenin-related proteins indicated consistent changes between TNKS2 andβ-catenin expression in lung cancer cells,whereas Axin displayed an opposite trend(P<0.05).CONCLUSION The obtained results revealed that TNKS2 may serve as an adverse prognostic factor and a potential therapeutic target in NSCLC.展开更多
This study compared tankyrase 1 expression and autophagy quantity between erectile dysfunction （ED） and non-ED rats＇ corpus cavernosum smooth muscle cells （CSMCs）. This study aslo explored the effect and possible...This study compared tankyrase 1 expression and autophagy quantity between erectile dysfunction （ED） and non-ED rats＇ corpus cavernosum smooth muscle cells （CSMCs）. This study aslo explored the effect and possible mechanism of tankyrase 1 on autophagy and cell proliferation in ageing ED rats＇ CSMCs. The intracavernous pres- sure and mean systemic arterial pressure were measured to investigate erectile function so that eight 24-month-old ED and eight 8-month-old male Wistar rats were choosed respectively. The rat CSMCs were isolated and cultured by enzyme digestion, in which tankyrase 1 expression and autophagy quantity were compared. Tankyrase 1 over-expression was induced with plasmid transfection by Lipofectamine^TM. The effect of tankyrase 1 overexpression on proliferation, autophagy and mTOR pathway in 24-month-old ED rats＇ CSMCs was measured by the cell growth curve in MTT assay, cell cycle analysis in flow cytometry （FCM）, key protein expression in Western blot, autophagy quantity in transmission electron microscopy, monodansylcadaverine staining and GFP-LC3 fluorescence. The primary CSMCs were confirmed by immunofluorescence, and the purity was 99.1% in FCM. Compared with that of 8-month-old rats, tankyrase 1 expression and autophagy quantity significantly decreased in 24-month-old ED rats＇ primary CSMCs （P 〈 0.01）. Tankyrase 1 overexpression significantly increased the growth rate （P 〈 0.05） and increased the S phase of cell cycle （P 〈 0.01）. The autophagosome quantity was remarkably increased （P 〈 0.01）, LC3-Ⅰ/Ⅱ and Beclin 1 were upregulated （P 〈 0.01 and P 〈 0.05）, and p-p70S6K （Thr389） was downregulated in 24-month-old ED rat CSMCs （P 〈 0.05）. In conclusion, Tankyrase 1 and autophagy decrease in the CSMCs from aging rats with ED, and tankyrase 1 may have a positive effect on proliferation by enhancing autophagy and regulating the mTOR signalling pathway.展开更多
The WNT/-catenin and phosphoinositide 3-kinase(PI3K/AKT) signaling cascades both have been implicated in the formation and progression of colorectal cancer.Oncogenic PI3K/AKT signaling suppresses the activity of forkh...The WNT/-catenin and phosphoinositide 3-kinase(PI3K/AKT) signaling cascades both have been implicated in the formation and progression of colorectal cancer.Oncogenic PI3K/AKT signaling suppresses the activity of forkhead box O3a(FOXO3a) transcription factor through phosphorylation leading to its nuclear exclusion.Inhibition of the PI3K/AKT signaling by PI3K or AKT inhibitors results in the translocation of FOXO3a to the nucleus,and is considered to be a promising therapeutic strategy for many cancers including colon cancer.Now,however,a new study in Nature Medicine has revealed a nuclear interaction of-catenin with FOXO3a as a promoter of metastatic progression in colon cancer.The work has important implications for the treatment of colon cancers,suggests a companion biomarker strategy to enable a personalized medicine approach,and offers an alternative therapeutic strategy to overcome resistance to PI3K and AKT inhibitors.展开更多
Poly(ADP-ribose)(PAR)is a highly negatively charged polymer.PAR is synthesized by poly(ADP-ribose)polymerases(PARPs)and is involved in the assembly and stabilization of macromolecular complexes.Here,the presence and p...Poly(ADP-ribose)(PAR)is a highly negatively charged polymer.PAR is synthesized by poly(ADP-ribose)polymerases(PARPs)and is involved in the assembly and stabilization of macromolecular complexes.Here,the presence and putative roles of poly(ADP-ribosyl)ation(PARylation)associated to adherens junctions(AJ)and the actin cytoskeleton in epithelial and Schwann cells,is reviewed.The hypothesis generated by analogy,stating that PAR is associated to AJ in other cell types,is postulated.According to this hypothesis,PAR associated to puncta adherentia in chemical synapses would participate in plasticity,learning and memory.In turn,PAR associated to fascia adherens in cardiomyocytes,would affect heart beating.PARP inhibitors are currently under development and clinical testing.Basic research in different tissues will probably influence their clinical uses.展开更多
基金Supported by Traditional Chinese Medicine Foundation of Zhejiang Province,No.2019ZA020。
文摘BACKGROUND Tankyrase 2(TNKS2)is a potential candidate molecular target for the prognosis and treatment of non-small cell lung cancer(NSCLC),but its biological functions are unclear.AIM To investigate the biological functions of TNKS2 in NSCLC.METHODS Using a lentiviral vector,we generated H647 model cells with TNKS2 knockdown by RNA interference and A549 model cells with TNKS2 overexpression by tran-sfection with a TNKS2 overexpressing plasmid.Increased and decreased exp-ression levels of TNKS2 in the two cell lines were verified using real-time reverse transcriptase-polymerase chain reaction and Western blot analyses.Cell apopto-sis,proliferation,and migration were determined using flow cytometry,carbo-xyfluorescein succinimidyl ester staining,and scratch assay,respectively.Im-munofluorescence staining was conducted to examine TNKS2 andβ-catenin ex-pression levels in the two transfected cell lines and the non-transfected cells.RESULTS TNKS2 mRNA and protein expression was significantly higher in the highly malignant NCI-H647 cells,while it remained at a low level in the less malignant A549 cells.Lentivirus-mediated overexpression of TNKS2 in A549 cells resulted in a 3-fold increase in gene expression and a 1.7-fold increase in protein expression(P<0.01).Conversely,shRNA interference targeting TNKS2 Led to an 8-fold decrease in gene expression and a 3-fold decrease in protein expression(P<0.01)in NCI-H647 cells.Furthermore,the cell apoptosis rate was significantly reduced(50%)and cell migration rate was increased(35%)in the TNKS2 overexpression group than in the control group(P<0.05).In contrast,shTNKS2 promoted apoptosis by more than one fold and reduced migration by 60%(P<0.05).Immunofluorescence analysis revealed enhanced nuclear localization ofβ-catenin fluorescence signal associated with high TNKS2 expression levels.Western blot analysis investigating TNKS2/β-catenin-related proteins indicated consistent changes between TNKS2 andβ-catenin expression in lung cancer cells,whereas Axin displayed an opposite trend(P<0.05).CONCLUSION The obtained results revealed that TNKS2 may serve as an adverse prognostic factor and a potential therapeutic target in NSCLC.
基金Acknowledgment We are grateful to Dr Tamotsu Yoshimori for providing the GFP-LC3 plasmid and Dr H. Seimiya for providing the tankyrase 1 plasmid. This study was supported by the National Natural Science Foundation of China （No. 30772285） and Beijing Municipal Commission of Science Technology, China （No. Z080507030808011）.
文摘This study compared tankyrase 1 expression and autophagy quantity between erectile dysfunction （ED） and non-ED rats＇ corpus cavernosum smooth muscle cells （CSMCs）. This study aslo explored the effect and possible mechanism of tankyrase 1 on autophagy and cell proliferation in ageing ED rats＇ CSMCs. The intracavernous pres- sure and mean systemic arterial pressure were measured to investigate erectile function so that eight 24-month-old ED and eight 8-month-old male Wistar rats were choosed respectively. The rat CSMCs were isolated and cultured by enzyme digestion, in which tankyrase 1 expression and autophagy quantity were compared. Tankyrase 1 over-expression was induced with plasmid transfection by Lipofectamine^TM. The effect of tankyrase 1 overexpression on proliferation, autophagy and mTOR pathway in 24-month-old ED rats＇ CSMCs was measured by the cell growth curve in MTT assay, cell cycle analysis in flow cytometry （FCM）, key protein expression in Western blot, autophagy quantity in transmission electron microscopy, monodansylcadaverine staining and GFP-LC3 fluorescence. The primary CSMCs were confirmed by immunofluorescence, and the purity was 99.1% in FCM. Compared with that of 8-month-old rats, tankyrase 1 expression and autophagy quantity significantly decreased in 24-month-old ED rats＇ primary CSMCs （P 〈 0.01）. Tankyrase 1 overexpression significantly increased the growth rate （P 〈 0.05） and increased the S phase of cell cycle （P 〈 0.01）. The autophagosome quantity was remarkably increased （P 〈 0.01）, LC3-Ⅰ/Ⅱ and Beclin 1 were upregulated （P 〈 0.01 and P 〈 0.05）, and p-p70S6K （Thr389） was downregulated in 24-month-old ED rat CSMCs （P 〈 0.05）. In conclusion, Tankyrase 1 and autophagy decrease in the CSMCs from aging rats with ED, and tankyrase 1 may have a positive effect on proliferation by enhancing autophagy and regulating the mTOR signalling pathway.
文摘The WNT/-catenin and phosphoinositide 3-kinase(PI3K/AKT) signaling cascades both have been implicated in the formation and progression of colorectal cancer.Oncogenic PI3K/AKT signaling suppresses the activity of forkhead box O3a(FOXO3a) transcription factor through phosphorylation leading to its nuclear exclusion.Inhibition of the PI3K/AKT signaling by PI3K or AKT inhibitors results in the translocation of FOXO3a to the nucleus,and is considered to be a promising therapeutic strategy for many cancers including colon cancer.Now,however,a new study in Nature Medicine has revealed a nuclear interaction of-catenin with FOXO3a as a promoter of metastatic progression in colon cancer.The work has important implications for the treatment of colon cancers,suggests a companion biomarker strategy to enable a personalized medicine approach,and offers an alternative therapeutic strategy to overcome resistance to PI3K and AKT inhibitors.
文摘Poly(ADP-ribose)(PAR)is a highly negatively charged polymer.PAR is synthesized by poly(ADP-ribose)polymerases(PARPs)and is involved in the assembly and stabilization of macromolecular complexes.Here,the presence and putative roles of poly(ADP-ribosyl)ation(PARylation)associated to adherens junctions(AJ)and the actin cytoskeleton in epithelial and Schwann cells,is reviewed.The hypothesis generated by analogy,stating that PAR is associated to AJ in other cell types,is postulated.According to this hypothesis,PAR associated to puncta adherentia in chemical synapses would participate in plasticity,learning and memory.In turn,PAR associated to fascia adherens in cardiomyocytes,would affect heart beating.PARP inhibitors are currently under development and clinical testing.Basic research in different tissues will probably influence their clinical uses.
