AIM:To investigate the role of pre-B-cell leukemia homeobox(PBX)3 in migration and invasion of colorectal cancer(CRC)cells.METHODS:We detected PBX3 expression in five cell lines and surgical specimens from 111 patient...AIM:To investigate the role of pre-B-cell leukemia homeobox(PBX)3 in migration and invasion of colorectal cancer(CRC)cells.METHODS:We detected PBX3 expression in five cell lines and surgical specimens from 111 patients with CRC using real-time reverse transcription-polymerase chain reaction.We forced expression of PBX3 in low metastatic HT-29 and SW480 cells and knocked down expression of PBX3 in highly metastatic LOVO and HCT-8 cells.Wound healing and Boyden chamber assays were used to detect cell migration and invasionafter altered expression of PBX3.Western blot was performed to detect the change of signaling molecule ERK1/2 following PBX3 overexpression.RESULTS:High level of PBX3 expression was correlated with the invasive potential of CRC cells,and significantly associated with lymph node invasion(P=0.02),distant metastasis(P=0.04),advanced TNM stage(P=0.03)and poor overall survival of patients(P<0.05).Ectopic expression of PBX3 in low metastatic cells was shown to promote migration and invasion,while inhibited PBX3 expression in highly metastatic cells suppressed migration and invasion.Furthermore,upregulation of phosphorylated extracellular signal-regulated kinase(ERK)1/2 was found to be one of the targeted molecules responsible for PBX3-induced CRC cell migration and invasion.CONCLUSION:PBX3 induces invasion and metastasis of CRC cells partially through activation of the MAPK/ERK signaling pathway.展开更多
BACKGROUND Ectopic expression of miRNAs promotes tumor development and progression.miRNA(miR)-320a is downregulated in many cancers,including gastric cancer(GC).However,the mechanism underlying its downregulation and ...BACKGROUND Ectopic expression of miRNAs promotes tumor development and progression.miRNA(miR)-320a is downregulated in many cancers,including gastric cancer(GC).However,the mechanism underlying its downregulation and the role of miR-320a in GC are unknown.AIM To determine expression and biological functions of miR-320a in GC and investigate the underlying molecular mechanisms.METHODS Quantitative real-time polymerase chain reaction(PCR)was used to determine expression of miR-320a in GC cell lines and tissues.TargetScanHuman7.1,miRDB,and microRNA.org were used to predict the possible targets of miR-320a,and a dual luciferase assay was used to confirm the findings.Western blotting was used to detect the protein levels of pre-B-cell leukemia homeobox 3(PBX3)in GC cells and tissue samples.Cell Counting Kit-8 proliferation,Transwell,wound healing,and apoptosis assays were performed to analyze the biological functions of miR-320a in GC cells.Methylation-specific PCR was used to analyze the methylation level of the miR-320a promoter CpG islands.5-Aza-2’-deoxycytidine(5-Aza-CdR)and trichostatin A(TSA)were used to treat GC cells.RESULTS miR-320a expression was lower in GC cell lines and tissues than in the normal gastric mucosa cell line GES-1 and matched adjacent normal tissues.miR-320a overexpression suppressed GC cell proliferation,invasion and migration,and induced apoptosis.PBX3 was a target of miR-320a in GC.The methylation level of the miR-320a promoter CpG islands was elevated and this was partly reversed by 5-Aza-CdR and TSA.CONCLUSION miR-320a acts as a tumor suppressor and inhibits malignant behavior of GC cells,partly by targeting PBX3.DNA methylation is an important mechanism associated with low expression of miR-320a.展开更多
基金Supported by Beijing Natural Science Foundation,No.5122012National Natural Science Foundation of China,No.81201964 and No.81330051
文摘AIM:To investigate the role of pre-B-cell leukemia homeobox(PBX)3 in migration and invasion of colorectal cancer(CRC)cells.METHODS:We detected PBX3 expression in five cell lines and surgical specimens from 111 patients with CRC using real-time reverse transcription-polymerase chain reaction.We forced expression of PBX3 in low metastatic HT-29 and SW480 cells and knocked down expression of PBX3 in highly metastatic LOVO and HCT-8 cells.Wound healing and Boyden chamber assays were used to detect cell migration and invasionafter altered expression of PBX3.Western blot was performed to detect the change of signaling molecule ERK1/2 following PBX3 overexpression.RESULTS:High level of PBX3 expression was correlated with the invasive potential of CRC cells,and significantly associated with lymph node invasion(P=0.02),distant metastasis(P=0.04),advanced TNM stage(P=0.03)and poor overall survival of patients(P<0.05).Ectopic expression of PBX3 in low metastatic cells was shown to promote migration and invasion,while inhibited PBX3 expression in highly metastatic cells suppressed migration and invasion.Furthermore,upregulation of phosphorylated extracellular signal-regulated kinase(ERK)1/2 was found to be one of the targeted molecules responsible for PBX3-induced CRC cell migration and invasion.CONCLUSION:PBX3 induces invasion and metastasis of CRC cells partially through activation of the MAPK/ERK signaling pathway.
基金Supported by the Natural Science Foundation of Liaoning Province,No.201602817
文摘BACKGROUND Ectopic expression of miRNAs promotes tumor development and progression.miRNA(miR)-320a is downregulated in many cancers,including gastric cancer(GC).However,the mechanism underlying its downregulation and the role of miR-320a in GC are unknown.AIM To determine expression and biological functions of miR-320a in GC and investigate the underlying molecular mechanisms.METHODS Quantitative real-time polymerase chain reaction(PCR)was used to determine expression of miR-320a in GC cell lines and tissues.TargetScanHuman7.1,miRDB,and microRNA.org were used to predict the possible targets of miR-320a,and a dual luciferase assay was used to confirm the findings.Western blotting was used to detect the protein levels of pre-B-cell leukemia homeobox 3(PBX3)in GC cells and tissue samples.Cell Counting Kit-8 proliferation,Transwell,wound healing,and apoptosis assays were performed to analyze the biological functions of miR-320a in GC cells.Methylation-specific PCR was used to analyze the methylation level of the miR-320a promoter CpG islands.5-Aza-2’-deoxycytidine(5-Aza-CdR)and trichostatin A(TSA)were used to treat GC cells.RESULTS miR-320a expression was lower in GC cell lines and tissues than in the normal gastric mucosa cell line GES-1 and matched adjacent normal tissues.miR-320a overexpression suppressed GC cell proliferation,invasion and migration,and induced apoptosis.PBX3 was a target of miR-320a in GC.The methylation level of the miR-320a promoter CpG islands was elevated and this was partly reversed by 5-Aza-CdR and TSA.CONCLUSION miR-320a acts as a tumor suppressor and inhibits malignant behavior of GC cells,partly by targeting PBX3.DNA methylation is an important mechanism associated with low expression of miR-320a.
