[Objectives]To study the inhibitory activity of two flavonoid glycosides isolated from Chlorophytum comosum Laxum R.Br on human nasopharyngeal carcinoma(NPC)cell line 5-8F in vitro and its mechanism.[Methods]The flavo...[Objectives]To study the inhibitory activity of two flavonoid glycosides isolated from Chlorophytum comosum Laxum R.Br on human nasopharyngeal carcinoma(NPC)cell line 5-8F in vitro and its mechanism.[Methods]The flavonoid glycosides were isolated and purified from the ethanol alcoholic extract of the roots of Liliaceae plant Chlorophytum comosum by silica gel column chromatography,macroporous resin column chromatography,Sephadex LH-20,and reverse column chromatography(ODS).The inhibitory activity of flavonoid glycosides on human nasopharyngeal carcinoma cells was analyzed by CCK-8 method,and the potential mechanism was preliminarily analyzed by molecular docking.[Results]Two flavonoid glycosides were identified as isovitexin 2″-0-rhamnoside and 7-2″-di-O-β-glucopyranosylisovitexin.Two flavonoid glycosides showed promising inhibitory effect on human nasopharyngeal carcinoma cell line 5-8F,with IC_(50) values of 24.8 and 27.5μmol/L,respectively.Molecular docking results showed that the potential targets of two flavonoid glycosides include CyclinD1,Bcl-2β-Catenin,ILK,TGF-β,in addition,two glycosides showed higher predicted binding affinity towards CyclinD1,which verifies the cytotoxicity of the two compounds on human nasopharyngeal carcinoma cell line 5-8F in vitro.[Conclusions]Two flavonoid glycosides are the active molecules in Chlorophytum comosum that can inhibit the proliferation of human nasopharyngeal carcinoma cells,and have the potential to be used in the research and development of anti nasopharyngeal carcinoma drugs.展开更多
Objective:To explore the role of circROBO1 in promoting the invasion of retinal Y79 cells by targeting KLF5 and its possible regulatory mechanism.Methods:RNase R enzyme digestion and qRT-PCR experiments were used to d...Objective:To explore the role of circROBO1 in promoting the invasion of retinal Y79 cells by targeting KLF5 and its possible regulatory mechanism.Methods:RNase R enzyme digestion and qRT-PCR experiments were used to detect the structural stability of circular circROBO1 in retinal Y79 cells;cytoplasmic and nuclear RNAs of retinal Y79 cells were extracted for localization analysis of circROBO1;The expression of circROBO1 in retinal Y79 cells were silenced by siRNA.The effect of circROBO1 on the migration and invasion ability of HT-29 cells was detected by scratch assay,Transwell cell invasion and migration assay.The target binding sites of circROBO1 and its downstream miRNA and that of miRNA and its downstream target gene KLF5 were predicted by CircInteractome and TargetScan online software respectively,and the target regulation relationship between them was verified by double luciferase reporter gene experiment.Western blot was used to detect the effect of siRNA silencing the expression of circROBO1 in Y79 cells on the expression of KLF5.Results:Compared with the control group without RNase R enzyme treatment,relative circROBO1 levels did not change significantly after treatment,while relative linear ROBO1 levels decreased significantly after treatment(t=16.18,P<0.05);the content of circROBO1 in the cytoplasm was significantly higher than that in the nucleus(P<0.05);compared with si-control group,the migration rate and the invasion and migration abilities of Transwell cells were all lower in the si-circROBO1 group(t=22.54,P<0.05);circROBO1 can adsorb miR-885-5p,and there is a target binding site between miR-885-5p and KLF5(t=11.39,P<0.05);compared with the si-control group,the KLF5 protein expression in the si-circROBO1 group was significantly decreased(t=17.26,P<0.05).Conclusions:circROBO1 promotes retinalY79 cell tumor invasion by targeting KLF5.展开更多
BACKGROUND Studies have shown that long non-coding RNAs(lncRNAs)play a key role in almost all key physiological and pathological processes,including different types of malignant tumors.Our previous lncRNA microarray r...BACKGROUND Studies have shown that long non-coding RNAs(lncRNAs)play a key role in almost all key physiological and pathological processes,including different types of malignant tumors.Our previous lncRNA microarray results have shown that lncRNA XLOC_001659 is upregulated in esophageal cancer(EC)tissues,with a fold change of 20.9 relative to normal esophageal tissues.But its effect and the molecular biological mechanisms on proliferation and invasion of EC cells remain unclear.AIM To investigate the effect of lncRNA XLOC_001659 on esophageal squamous cell carcinoma(ESCC)cells and explore the molecular biological mechanisms involved.METHODS RT-qPCR assay was used to quantify the expression levels of lncRNAXLOC-001659 and miR-490-5p.The proliferative capacity of the cells was determined using CCK8 and colony formation assays,and the effect of lncRNAXLOC-001659 on the invasion of ESCC cells was determined by Transwell assay.Dualluciferase reporter assay was used to detect the target genes of lncRNAXLOC-001659 and miR-490-5p.RESULTS The results of RT-qPCR showed that the expression of lncRNAXLOC_001659 was upregulated in ESCC cells.CCK-8 assay showed that knockdown of lncRNAXLOC_001659 significantly inhibited ESCC cell proliferation.Colony formation and Transwell invasion assays showed that knockdown of lncRNAXLOC_001659 or overexpression of miR-490-5p significantly inhibited ESCC cell growth and invasion.Furthermore,lncRNAXLOC_001659 acts as an endogenous sponge by competitively binding to miR-490-5p to downregulate miR-490-5p.Further results confirmed that miR-490-5p targeted PIK3CA,and the recovery of PIK3CA rescued lncRNAXLOC_001659 knockdown or miR-490-5p overexpression-mediated inhibition of cell proliferation and invasion,which suggested the presence of an lncRNAXLOC_001659/miR-490-5p/PIK3CA regulatory axis.CONCLUSION Knockdown of lncRNA XLOC_001659 inhibits proliferation and invasion of ESCC cells via regulation of miR-490-5p/PIK3CA,suggesting that it may play a role in ESCC tumorigenesis and progression.展开更多
基金Supported by Youth Fund Project of Zhaoqing University(QZ202235)Zhaoqing Science and Technology Plan Project(2022040311011).
文摘[Objectives]To study the inhibitory activity of two flavonoid glycosides isolated from Chlorophytum comosum Laxum R.Br on human nasopharyngeal carcinoma(NPC)cell line 5-8F in vitro and its mechanism.[Methods]The flavonoid glycosides were isolated and purified from the ethanol alcoholic extract of the roots of Liliaceae plant Chlorophytum comosum by silica gel column chromatography,macroporous resin column chromatography,Sephadex LH-20,and reverse column chromatography(ODS).The inhibitory activity of flavonoid glycosides on human nasopharyngeal carcinoma cells was analyzed by CCK-8 method,and the potential mechanism was preliminarily analyzed by molecular docking.[Results]Two flavonoid glycosides were identified as isovitexin 2″-0-rhamnoside and 7-2″-di-O-β-glucopyranosylisovitexin.Two flavonoid glycosides showed promising inhibitory effect on human nasopharyngeal carcinoma cell line 5-8F,with IC_(50) values of 24.8 and 27.5μmol/L,respectively.Molecular docking results showed that the potential targets of two flavonoid glycosides include CyclinD1,Bcl-2β-Catenin,ILK,TGF-β,in addition,two glycosides showed higher predicted binding affinity towards CyclinD1,which verifies the cytotoxicity of the two compounds on human nasopharyngeal carcinoma cell line 5-8F in vitro.[Conclusions]Two flavonoid glycosides are the active molecules in Chlorophytum comosum that can inhibit the proliferation of human nasopharyngeal carcinoma cells,and have the potential to be used in the research and development of anti nasopharyngeal carcinoma drugs.
基金Key Project of Hunan Provincial Department of Education(No.21A0285)。
文摘Objective:To explore the role of circROBO1 in promoting the invasion of retinal Y79 cells by targeting KLF5 and its possible regulatory mechanism.Methods:RNase R enzyme digestion and qRT-PCR experiments were used to detect the structural stability of circular circROBO1 in retinal Y79 cells;cytoplasmic and nuclear RNAs of retinal Y79 cells were extracted for localization analysis of circROBO1;The expression of circROBO1 in retinal Y79 cells were silenced by siRNA.The effect of circROBO1 on the migration and invasion ability of HT-29 cells was detected by scratch assay,Transwell cell invasion and migration assay.The target binding sites of circROBO1 and its downstream miRNA and that of miRNA and its downstream target gene KLF5 were predicted by CircInteractome and TargetScan online software respectively,and the target regulation relationship between them was verified by double luciferase reporter gene experiment.Western blot was used to detect the effect of siRNA silencing the expression of circROBO1 in Y79 cells on the expression of KLF5.Results:Compared with the control group without RNase R enzyme treatment,relative circROBO1 levels did not change significantly after treatment,while relative linear ROBO1 levels decreased significantly after treatment(t=16.18,P<0.05);the content of circROBO1 in the cytoplasm was significantly higher than that in the nucleus(P<0.05);compared with si-control group,the migration rate and the invasion and migration abilities of Transwell cells were all lower in the si-circROBO1 group(t=22.54,P<0.05);circROBO1 can adsorb miR-885-5p,and there is a target binding site between miR-885-5p and KLF5(t=11.39,P<0.05);compared with the si-control group,the KLF5 protein expression in the si-circROBO1 group was significantly decreased(t=17.26,P<0.05).Conclusions:circROBO1 promotes retinalY79 cell tumor invasion by targeting KLF5.
基金Supported by Excellent Youth Development Fund of Zhengzhou University,No.1621328001
文摘BACKGROUND Studies have shown that long non-coding RNAs(lncRNAs)play a key role in almost all key physiological and pathological processes,including different types of malignant tumors.Our previous lncRNA microarray results have shown that lncRNA XLOC_001659 is upregulated in esophageal cancer(EC)tissues,with a fold change of 20.9 relative to normal esophageal tissues.But its effect and the molecular biological mechanisms on proliferation and invasion of EC cells remain unclear.AIM To investigate the effect of lncRNA XLOC_001659 on esophageal squamous cell carcinoma(ESCC)cells and explore the molecular biological mechanisms involved.METHODS RT-qPCR assay was used to quantify the expression levels of lncRNAXLOC-001659 and miR-490-5p.The proliferative capacity of the cells was determined using CCK8 and colony formation assays,and the effect of lncRNAXLOC-001659 on the invasion of ESCC cells was determined by Transwell assay.Dualluciferase reporter assay was used to detect the target genes of lncRNAXLOC-001659 and miR-490-5p.RESULTS The results of RT-qPCR showed that the expression of lncRNAXLOC_001659 was upregulated in ESCC cells.CCK-8 assay showed that knockdown of lncRNAXLOC_001659 significantly inhibited ESCC cell proliferation.Colony formation and Transwell invasion assays showed that knockdown of lncRNAXLOC_001659 or overexpression of miR-490-5p significantly inhibited ESCC cell growth and invasion.Furthermore,lncRNAXLOC_001659 acts as an endogenous sponge by competitively binding to miR-490-5p to downregulate miR-490-5p.Further results confirmed that miR-490-5p targeted PIK3CA,and the recovery of PIK3CA rescued lncRNAXLOC_001659 knockdown or miR-490-5p overexpression-mediated inhibition of cell proliferation and invasion,which suggested the presence of an lncRNAXLOC_001659/miR-490-5p/PIK3CA regulatory axis.CONCLUSION Knockdown of lncRNA XLOC_001659 inhibits proliferation and invasion of ESCC cells via regulation of miR-490-5p/PIK3CA,suggesting that it may play a role in ESCC tumorigenesis and progression.
基金This work was supported by the NNSFC(No.50273024)the Natural Foundation of Jiangsu Province(No.BK2002041 and BK2003031)the Foundation of Jiangsu Province Education Committee(No.02KJB430001 and 03KJB150115).
