BACKGROUND Diabetic skin ulcers,a significant global healthcare burden,are mainly caused by the inhibition of cell proliferation and impaired angiogenesis.XB130 is an adaptor protein that regulates cell proliferation ...BACKGROUND Diabetic skin ulcers,a significant global healthcare burden,are mainly caused by the inhibition of cell proliferation and impaired angiogenesis.XB130 is an adaptor protein that regulates cell proliferation and migration.However,the role of XB130 in the development of diabetic skin ulcers remains unclear.AIM To investigate whether XB130 can regulate the inhibition of proliferation and vascular damage induced by high glucose.Additionally,we aim to determine whether XB130 is involved in the healing process of diabetic skin ulcers,along with its molecular mechanisms.METHODS We conducted RNA-sequencing analysis to identify the key genes involved in diabetic skin ulcers.We investigated the effects of XB130 on wound healing using histological analyses.In addition,we used reverse transcription-quantitative polymerase chain reaction,Western blot,terminal deoxynucleotidyl transferasemediated dUTP nick end labeling staining,immunofluorescence,wound healing,and tubule formation experiments to investigate their effects on cellular processes in human umbilical vein endothelial cells(HUVECs)stimulated with high glucose.Finally,we performed functional analysis to elucidate the molecular mechanisms underlying diabetic skin ulcers.RESULTS RNA-sequencing analysis showed that the expression of XB130 was up-regulated in the tissues of diabetic skin ulcers.Knockdown of XB130 promoted the healing of skin wounds in mice,leading to an accelerated wound healing process and shortened wound healing time.At the cellular level,knockdown of XB130 alleviated high glucose-induced inhibition of cell proliferation and angiogenic impairment in HUVECs.Inhibition of the PI3K/Akt pathway removed the proliferative effects and endothelial protection mediated by XB130.CONCLUSION The findings of this study indicated that the expression of XB130 is up-regulated in high glucose-stimulated diabetic skin ulcers and HUVECs.Knockdown of XB130 promotes cell proliferation and angiogenesis via the PI3K/Akt signalling pathway,which accelerates the healing of diabetic skin ulcers.展开更多
Background：XB 130 is a recently discovered adaptor protein that is highly expressed in many malignant tumors,but few studies have investigated its role in hepatocellular carcinoma （HCC）.Therefore,this study explore...Background：XB 130 is a recently discovered adaptor protein that is highly expressed in many malignant tumors,but few studies have investigated its role in hepatocellular carcinoma （HCC）.Therefore,this study explored the relationship between this protein and liver cancer and investigated its molecular mechanism of action.Methods：The expression of XB 130 between HCC tissues and adjacent nontumor tissues was compared by real-time polymerase chain reaction,immunochemistry,and Western blotting.XB130 silencing was performed using small hairpin RNA.The effect of silencing XB130 was examined using Cell Counting Kit-8,colony assay,wound healing assay,and cell cycle analysis.Results：We found that XB 130 was highly expressed in HCC tissues （cancer tissues vs.adjacent tissues：0.23 ± 0.02 vs.0.17 ± 0.02,P 〈 0.05） and liver cancer cell lines,particularly MHCC97H and HepG2 （MHCC97H and HepG2 vs.normal liver cell line LO-2：2.35 ± 0.26 and 2.04 ± 0.04 vs.1.00 ± 0.04,respectively,all P 〈 0.05）.The Cell Counting Kit-8 assay,colony formation assay,and xenograft model in nude mice showed that silencing XB130 inhibited cell proliferative ability both in vivo and in vitro,with flow cytometry demonstrating that the cells were arrested in the G0/G1 phase in HepG2 （HepG2 XB130-silenced group [shA] vs.HepG2 scramble group [NA]：74.32 ± 5.86% vs.60.21 ± 3.07%,P 〈 0.05） and that the number of G2/M phase cells was decreased （HepG2 shA vs.HepG2 NA：8.06 ± 2.41% vs.18.36 ± 4.42%,P 〈 0.05）.Furthermore,the cell invasion and migration abilities were impaired,and the levels of the epithelial-mesenchymal transition-related indicators vimentin and N-cadherin were decreased,although the level of E-cadherin was increased after silencingXB130.Western blotting showed that the levels of phosphorylated phosphoinositide 3-kinase （PI3K） and phospho-protein kinase B （p-Akt） also increased,although the level of phosphorylated phosphatase and tensin homolog increased,indicating that XB 130 activated the PI3K/ Akt pathway.Furthermore,we found that a reduction in XB130 increased liver cancer cell sensitivity to tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis.Conclusions：Our findings suggest that XB130 might be used as a predictor of liver cancer as well as one of the targets for its treatment.展开更多
基金the National Natural Science Foundation of China,No.82272355Shanghai Science and Technology Committee,No.21410750500.
文摘BACKGROUND Diabetic skin ulcers,a significant global healthcare burden,are mainly caused by the inhibition of cell proliferation and impaired angiogenesis.XB130 is an adaptor protein that regulates cell proliferation and migration.However,the role of XB130 in the development of diabetic skin ulcers remains unclear.AIM To investigate whether XB130 can regulate the inhibition of proliferation and vascular damage induced by high glucose.Additionally,we aim to determine whether XB130 is involved in the healing process of diabetic skin ulcers,along with its molecular mechanisms.METHODS We conducted RNA-sequencing analysis to identify the key genes involved in diabetic skin ulcers.We investigated the effects of XB130 on wound healing using histological analyses.In addition,we used reverse transcription-quantitative polymerase chain reaction,Western blot,terminal deoxynucleotidyl transferasemediated dUTP nick end labeling staining,immunofluorescence,wound healing,and tubule formation experiments to investigate their effects on cellular processes in human umbilical vein endothelial cells(HUVECs)stimulated with high glucose.Finally,we performed functional analysis to elucidate the molecular mechanisms underlying diabetic skin ulcers.RESULTS RNA-sequencing analysis showed that the expression of XB130 was up-regulated in the tissues of diabetic skin ulcers.Knockdown of XB130 promoted the healing of skin wounds in mice,leading to an accelerated wound healing process and shortened wound healing time.At the cellular level,knockdown of XB130 alleviated high glucose-induced inhibition of cell proliferation and angiogenic impairment in HUVECs.Inhibition of the PI3K/Akt pathway removed the proliferative effects and endothelial protection mediated by XB130.CONCLUSION The findings of this study indicated that the expression of XB130 is up-regulated in high glucose-stimulated diabetic skin ulcers and HUVECs.Knockdown of XB130 promotes cell proliferation and angiogenesis via the PI3K/Akt signalling pathway,which accelerates the healing of diabetic skin ulcers.
文摘Background：XB 130 is a recently discovered adaptor protein that is highly expressed in many malignant tumors,but few studies have investigated its role in hepatocellular carcinoma （HCC）.Therefore,this study explored the relationship between this protein and liver cancer and investigated its molecular mechanism of action.Methods：The expression of XB 130 between HCC tissues and adjacent nontumor tissues was compared by real-time polymerase chain reaction,immunochemistry,and Western blotting.XB130 silencing was performed using small hairpin RNA.The effect of silencing XB130 was examined using Cell Counting Kit-8,colony assay,wound healing assay,and cell cycle analysis.Results：We found that XB 130 was highly expressed in HCC tissues （cancer tissues vs.adjacent tissues：0.23 ± 0.02 vs.0.17 ± 0.02,P 〈 0.05） and liver cancer cell lines,particularly MHCC97H and HepG2 （MHCC97H and HepG2 vs.normal liver cell line LO-2：2.35 ± 0.26 and 2.04 ± 0.04 vs.1.00 ± 0.04,respectively,all P 〈 0.05）.The Cell Counting Kit-8 assay,colony formation assay,and xenograft model in nude mice showed that silencing XB130 inhibited cell proliferative ability both in vivo and in vitro,with flow cytometry demonstrating that the cells were arrested in the G0/G1 phase in HepG2 （HepG2 XB130-silenced group [shA] vs.HepG2 scramble group [NA]：74.32 ± 5.86% vs.60.21 ± 3.07%,P 〈 0.05） and that the number of G2/M phase cells was decreased （HepG2 shA vs.HepG2 NA：8.06 ± 2.41% vs.18.36 ± 4.42%,P 〈 0.05）.Furthermore,the cell invasion and migration abilities were impaired,and the levels of the epithelial-mesenchymal transition-related indicators vimentin and N-cadherin were decreased,although the level of E-cadherin was increased after silencingXB130.Western blotting showed that the levels of phosphorylated phosphoinositide 3-kinase （PI3K） and phospho-protein kinase B （p-Akt） also increased,although the level of phosphorylated phosphatase and tensin homolog increased,indicating that XB 130 activated the PI3K/ Akt pathway.Furthermore,we found that a reduction in XB130 increased liver cancer cell sensitivity to tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis.Conclusions：Our findings suggest that XB130 might be used as a predictor of liver cancer as well as one of the targets for its treatment.
