BACKGROUND Liver cancer is one of the most highly malignant cancers,characterized by easy metastasis and chemoradiotherapy resistance.Emerging evidence indicates that long noncoding RNAs(LncRNAs),including Lnc524369,a...BACKGROUND Liver cancer is one of the most highly malignant cancers,characterized by easy metastasis and chemoradiotherapy resistance.Emerging evidence indicates that long noncoding RNAs(LncRNAs),including Lnc524369,are highly involved in the initiation,progression,radioresistance,and chemoresistance of hepatocellular carcinoma(HCC).However,the function of Lnc524369 remains unclear.AIM To explore the function of Lnc524369 in HCC.METHODS To investigate the effect of Lnc524369,tissue from 41 HCC patients were analyzed using CCK8,migration,and invasion assays.Lnc524369 and YWHAZ(also named 14-3-3ζ)mRNA were detected by qPCR,and YWHAZ and RAF1 proteins were detected by western blot in liver cancer cell lines and human HCC tissues.The Cancer Cell Line Encyclopedia(CCLE)databases,STRING database,Human Protein Atlas database,and the TCGA database were used for bioinformatic analysis.RESULTS Lnc524369 was significantly upregulated in the nucleus of liver cancer cells and human HCC tissues.Overexpression of Lnc524369 was associated with the proliferation,migration,and invasion of liver cancer cells.YWHAZ and RAF1 proteins and YWHAZ mRNA were overexpressed in liver cancer,which could be attenuated by overexpression of Lnc524369.Lnc524369 and its downstream target YWHAZ and RAF1 proteins were negatively associated with overall survival time.CONCLUSION Lnc524369 might be a promising target of HCC as it can enhance liver cancer progression and decrease the overall survival time of HCC by activating the YWHAZ/RAF1 pathway.展开更多
Objective To investigate the therapeutic effect of gentisic acid(GA)on rheumatoid arthritis(RA)based on the miR-19b-3p/RAF1 axis.Methods The cell counting kit-8 method was used to detect the growth inhibitory effect o...Objective To investigate the therapeutic effect of gentisic acid(GA)on rheumatoid arthritis(RA)based on the miR-19b-3p/RAF1 axis.Methods The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells,and the drug concentration of GA was determined in the experiment.The quantificational real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-19b-3p and RAF1.RAF1,extracellular regulated protein kinases1/2(ERK1/2)and phospho-ERK1/2(p-ERK1/2)were examined by Western blotting.Three methods(dual-luciferase assay,qRT-PCR and Western blot analysis)were used to verify miR-19b-3p targeting RAF1.Flow cytometry was performed to detect MH7A cell apoptosis.Transwell and wound healing assays were used to determine the invasion and migration capacities of MH7A cells.Results The growth of MH7A cells was gradually inhibited with increasing GA concentration.When the GA concentration exceeded 80 mmol/L,GA was significantly cytotoxic to MH7A cells,so the half maximal inhibitory concentration of GA for MH7A cells was calculated as 67.019 mmol/L.GA upregulated miR-19b-3p expression,downregulated RAF1 expression,inhibited ERK1/2 phosphorylation,induced MH7A cell apoptosis and suppressed MH7A cell invasion and migration(P<0.05 or P<0.01).RAF1 was identified as the target of miR-19b-3p and reversed inhibitory effects on miR-19b-3p expression(P<0.05 or P<0.01).The miR-19b-3p inhibitor upregulated RAF1 expression and ERK1/2 phosphorylation,suppressed MH7A cell apoptosis and induced MH7A cell invasion and migration(P<0.01).Conclusion GA regulated miR-19b-3p/RAF1 axis to mediate ERK pathway and inhibit the development of RA.展开更多
目的应用全外显子测序技术对疑似Noonan综合征(NS)的危重早产儿及其父母进行基因诊断,探讨基因型与临床表型之间的关系。方法收集患儿及其父母的外周血标本及临床资料,应用Agilent Sure Select Human All Exome V5试剂盒进行全基因组外...目的应用全外显子测序技术对疑似Noonan综合征(NS)的危重早产儿及其父母进行基因诊断,探讨基因型与临床表型之间的关系。方法收集患儿及其父母的外周血标本及临床资料,应用Agilent Sure Select Human All Exome V5试剂盒进行全基因组外显子捕获,Illumina 550测序仪进行双端高通量测序,Sanger测序验证家系成员以明确突变遗传方式。结果全外显子测序发现NS患儿RAF1基因杂合错义突变c. 770C>T,p. S257L、Sanger测序验证家系成员为新发突变。结合患儿特殊面容、双侧心房及右室增大、中度肺动脉高压(PAH)等临床症状诊断为Noonan综合征。结论应用全外显子测序技术能够快速发现Noonan综合征基因突变,对重症新生儿病例的明确诊断及临床遗传咨询具有重要意义。展开更多
基金the Medical and Health Science and Technology Project of Zhejiang Province(No.2021KY043)Chinese Foundation for Hepatitis Prevention and Control-TianQing Liver Disease Research Fund Subject(No.TQGB2020168).
文摘BACKGROUND Liver cancer is one of the most highly malignant cancers,characterized by easy metastasis and chemoradiotherapy resistance.Emerging evidence indicates that long noncoding RNAs(LncRNAs),including Lnc524369,are highly involved in the initiation,progression,radioresistance,and chemoresistance of hepatocellular carcinoma(HCC).However,the function of Lnc524369 remains unclear.AIM To explore the function of Lnc524369 in HCC.METHODS To investigate the effect of Lnc524369,tissue from 41 HCC patients were analyzed using CCK8,migration,and invasion assays.Lnc524369 and YWHAZ(also named 14-3-3ζ)mRNA were detected by qPCR,and YWHAZ and RAF1 proteins were detected by western blot in liver cancer cell lines and human HCC tissues.The Cancer Cell Line Encyclopedia(CCLE)databases,STRING database,Human Protein Atlas database,and the TCGA database were used for bioinformatic analysis.RESULTS Lnc524369 was significantly upregulated in the nucleus of liver cancer cells and human HCC tissues.Overexpression of Lnc524369 was associated with the proliferation,migration,and invasion of liver cancer cells.YWHAZ and RAF1 proteins and YWHAZ mRNA were overexpressed in liver cancer,which could be attenuated by overexpression of Lnc524369.Lnc524369 and its downstream target YWHAZ and RAF1 proteins were negatively associated with overall survival time.CONCLUSION Lnc524369 might be a promising target of HCC as it can enhance liver cancer progression and decrease the overall survival time of HCC by activating the YWHAZ/RAF1 pathway.
基金Supported by a grant from Zunyi Science and Technology Bureau,China(No.[2019]194)。
文摘Objective To investigate the therapeutic effect of gentisic acid(GA)on rheumatoid arthritis(RA)based on the miR-19b-3p/RAF1 axis.Methods The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells,and the drug concentration of GA was determined in the experiment.The quantificational real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-19b-3p and RAF1.RAF1,extracellular regulated protein kinases1/2(ERK1/2)and phospho-ERK1/2(p-ERK1/2)were examined by Western blotting.Three methods(dual-luciferase assay,qRT-PCR and Western blot analysis)were used to verify miR-19b-3p targeting RAF1.Flow cytometry was performed to detect MH7A cell apoptosis.Transwell and wound healing assays were used to determine the invasion and migration capacities of MH7A cells.Results The growth of MH7A cells was gradually inhibited with increasing GA concentration.When the GA concentration exceeded 80 mmol/L,GA was significantly cytotoxic to MH7A cells,so the half maximal inhibitory concentration of GA for MH7A cells was calculated as 67.019 mmol/L.GA upregulated miR-19b-3p expression,downregulated RAF1 expression,inhibited ERK1/2 phosphorylation,induced MH7A cell apoptosis and suppressed MH7A cell invasion and migration(P<0.05 or P<0.01).RAF1 was identified as the target of miR-19b-3p and reversed inhibitory effects on miR-19b-3p expression(P<0.05 or P<0.01).The miR-19b-3p inhibitor upregulated RAF1 expression and ERK1/2 phosphorylation,suppressed MH7A cell apoptosis and induced MH7A cell invasion and migration(P<0.01).Conclusion GA regulated miR-19b-3p/RAF1 axis to mediate ERK pathway and inhibit the development of RA.
文摘目的应用全外显子测序技术对疑似Noonan综合征(NS)的危重早产儿及其父母进行基因诊断,探讨基因型与临床表型之间的关系。方法收集患儿及其父母的外周血标本及临床资料,应用Agilent Sure Select Human All Exome V5试剂盒进行全基因组外显子捕获,Illumina 550测序仪进行双端高通量测序,Sanger测序验证家系成员以明确突变遗传方式。结果全外显子测序发现NS患儿RAF1基因杂合错义突变c. 770C>T,p. S257L、Sanger测序验证家系成员为新发突变。结合患儿特殊面容、双侧心房及右室增大、中度肺动脉高压(PAH)等临床症状诊断为Noonan综合征。结论应用全外显子测序技术能够快速发现Noonan综合征基因突变,对重症新生儿病例的明确诊断及临床遗传咨询具有重要意义。
