BACKGROUNDGastric cancer(GC)has a high prevalence and mortality overall.GEN1 is associatedwith abnormal centrosome amplification,DNA damage and increasedapoptosis.To date,little is known about the function and mechani...BACKGROUNDGastric cancer(GC)has a high prevalence and mortality overall.GEN1 is associatedwith abnormal centrosome amplification,DNA damage and increasedapoptosis.To date,little is known about the function and mechanism of GEN1 inGC.AIMTo explore the cellular processes associated with GC will help to elucidate themechanism of the occurrence and development of GC and discover potentialtherapeutic targets.METHODSThe detection of GEN1 expression at mRNA and protein levels was done by realtimequantitative polymerase chain reaction and western blotting.The function ofGEN1 was verified by loss-of-function experiments in AGS cells.The genes coexpressedwith GEN1 were searched from the stomach adenocarcinomas(STAD)data in The Cancer Genome Atlas database.Kyoto Encyclopedia of Genes andGenomes(KEGG)enrichment analysis of the genes co-expressed with GEN1 tofurther identify the pathways involved in GEN1.Rescue experiments usingferroptosis inhibitor ferrostatin-1 and chemotherapeutic sensitivity assays withcisplatin were also performed.RESULTSSignificant up-regulation of GEN1 was observed in GC cell lines AGS and MGC-803.Inhibition of GEN1 induced cell apoptosis and decreased cell proliferation,cycle progression,migration in AGS cells.There were 264 genes co-expressedwith GEN1 in STAD cohort(r>0.4,P<0.001).KEGG enrichment analysis showed that GEN1 might be associated with the cell cycle,Fanconi anemia pathway,homologous recombination,oocytemeiosis and cellular senescence in GC.Furthermore,CCNA2,CCNB1,CCNB2,cyclin-dependent kinase(CDK)1,CDK2 and polo-like kinase 1 protein levels were lower in GEN1-knockdown AGS cells,manifesting that GEN1 wasassociated with the cell cycle pathway in AGS cells.Downregulation of GEN1 decreased adenosine triphosphatecontent and elevated reactive oxygen species in AGS cells,suggesting that GEN1 silencing led to mitochondrialdysfunction in AGS cells.In addition,GEN1 silencing caused an overt decrease in FTH1 and GPX4 protein levelsand a significant elevation in ACSL4 protein levels,implying that GEN1 silencing promoted AGS cell ferroptosis.Treatment with ferrostatin-1 rescued cell viability loss induced by GEN1 knockdown,confirming ferroptosis as akey death mechanism.Additionally,GEN1-deficient AGS cells showed enhanced sensitivity to cisplatin,with asignificantly reduced half-maximal inhibitory concentration compared to control cells.CONCLUSIONGEN1 promotes GC cell proliferation and migration while suppressing apoptosis and ferroptosis.Targeting GEN1not only disrupts mitochondrial function and cell cycle progression but also sensitizes GC cells to ferroptosis andchemotherapy.These findings highlight GEN1 as a potential therapeutic target for enhancing treatment efficacy ingastric cancer.展开更多
On January 23,we arrived in Shuanghu County,Nagqu City,Xizang.With an average altitude of 5,000 meters,this is the heart of Changtang no man's land.Along the journey,Tibetan wild donkeys and Tibetan antelopes were...On January 23,we arrived in Shuanghu County,Nagqu City,Xizang.With an average altitude of 5,000 meters,this is the heart of Changtang no man's land.Along the journey,Tibetan wild donkeys and Tibetan antelopes were often seen on both sides of the highway instead of traces of human presence,and sand and dust kept hitting our van.As the night fell,the highest county seat in the world finally came into view.展开更多
Zur Aufklarung der Rolle dreier DNA Reparaturenzyme in der Hepatokarzino genese haben wir die hMTH 1,hOGG 1 und hMYHα mRNA Expression mittels RT/semi quantitativer Echtzeit PCR und der 8 OHdG Gehalt mittels H...Zur Aufklarung der Rolle dreier DNA Reparaturenzyme in der Hepatokarzino genese haben wir die hMTH 1,hOGG 1 und hMYHα mRNA Expression mittels RT/semi quantitativer Echtzeit PCR und der 8 OHdG Gehalt mittels HPLC/ECD in Tumor und Peritumorgewebe von 21 Patienten mit hepatozellularem Karzinom (HCC ) bestimmt. Es wurde gezeigt, da der 8 OHdG Gehalt im Peritumorgewebe signif ikant hoher als im Tumorgeweb e war ( P =0.006) und mit dem Entzundungsgrad korrelierte. Die hMTH 1 E xpression war im Tumorgewebe gegenuber dem Peritumorgewebe erhoht ( P =0.014). Umgekehrt war die hMYHα Expression im Peritumorgewebe signifikant hoher als im Tumorgewebe ( P =0.039). Fur die hOGG 1 Expression wurde kein deutlicher Unterschied zwischen Tumor und Peritumorgewebe beobachte t. Eine deutliche lineare Korrelation zwischen der hMTH 1 und der hOGG 1 Exp ression wurde sowohl in Tumor ( r =0.809, P < 0.001) als auch in Peritum orgewebe ( r =0.883, P <0.001) gefunden. Diese Daten sprechen fur ein e reaktive und gegen eine pathogenetische Rolle der untersuchten DNA Reparature nzyme in der Hepatokarzinogenese.展开更多
文摘BACKGROUNDGastric cancer(GC)has a high prevalence and mortality overall.GEN1 is associatedwith abnormal centrosome amplification,DNA damage and increasedapoptosis.To date,little is known about the function and mechanism of GEN1 inGC.AIMTo explore the cellular processes associated with GC will help to elucidate themechanism of the occurrence and development of GC and discover potentialtherapeutic targets.METHODSThe detection of GEN1 expression at mRNA and protein levels was done by realtimequantitative polymerase chain reaction and western blotting.The function ofGEN1 was verified by loss-of-function experiments in AGS cells.The genes coexpressedwith GEN1 were searched from the stomach adenocarcinomas(STAD)data in The Cancer Genome Atlas database.Kyoto Encyclopedia of Genes andGenomes(KEGG)enrichment analysis of the genes co-expressed with GEN1 tofurther identify the pathways involved in GEN1.Rescue experiments usingferroptosis inhibitor ferrostatin-1 and chemotherapeutic sensitivity assays withcisplatin were also performed.RESULTSSignificant up-regulation of GEN1 was observed in GC cell lines AGS and MGC-803.Inhibition of GEN1 induced cell apoptosis and decreased cell proliferation,cycle progression,migration in AGS cells.There were 264 genes co-expressedwith GEN1 in STAD cohort(r>0.4,P<0.001).KEGG enrichment analysis showed that GEN1 might be associated with the cell cycle,Fanconi anemia pathway,homologous recombination,oocytemeiosis and cellular senescence in GC.Furthermore,CCNA2,CCNB1,CCNB2,cyclin-dependent kinase(CDK)1,CDK2 and polo-like kinase 1 protein levels were lower in GEN1-knockdown AGS cells,manifesting that GEN1 wasassociated with the cell cycle pathway in AGS cells.Downregulation of GEN1 decreased adenosine triphosphatecontent and elevated reactive oxygen species in AGS cells,suggesting that GEN1 silencing led to mitochondrialdysfunction in AGS cells.In addition,GEN1 silencing caused an overt decrease in FTH1 and GPX4 protein levelsand a significant elevation in ACSL4 protein levels,implying that GEN1 silencing promoted AGS cell ferroptosis.Treatment with ferrostatin-1 rescued cell viability loss induced by GEN1 knockdown,confirming ferroptosis as akey death mechanism.Additionally,GEN1-deficient AGS cells showed enhanced sensitivity to cisplatin,with asignificantly reduced half-maximal inhibitory concentration compared to control cells.CONCLUSIONGEN1 promotes GC cell proliferation and migration while suppressing apoptosis and ferroptosis.Targeting GEN1not only disrupts mitochondrial function and cell cycle progression but also sensitizes GC cells to ferroptosis andchemotherapy.These findings highlight GEN1 as a potential therapeutic target for enhancing treatment efficacy ingastric cancer.
文摘On January 23,we arrived in Shuanghu County,Nagqu City,Xizang.With an average altitude of 5,000 meters,this is the heart of Changtang no man's land.Along the journey,Tibetan wild donkeys and Tibetan antelopes were often seen on both sides of the highway instead of traces of human presence,and sand and dust kept hitting our van.As the night fell,the highest county seat in the world finally came into view.
文摘Zur Aufklarung der Rolle dreier DNA Reparaturenzyme in der Hepatokarzino genese haben wir die hMTH 1,hOGG 1 und hMYHα mRNA Expression mittels RT/semi quantitativer Echtzeit PCR und der 8 OHdG Gehalt mittels HPLC/ECD in Tumor und Peritumorgewebe von 21 Patienten mit hepatozellularem Karzinom (HCC ) bestimmt. Es wurde gezeigt, da der 8 OHdG Gehalt im Peritumorgewebe signif ikant hoher als im Tumorgeweb e war ( P =0.006) und mit dem Entzundungsgrad korrelierte. Die hMTH 1 E xpression war im Tumorgewebe gegenuber dem Peritumorgewebe erhoht ( P =0.014). Umgekehrt war die hMYHα Expression im Peritumorgewebe signifikant hoher als im Tumorgewebe ( P =0.039). Fur die hOGG 1 Expression wurde kein deutlicher Unterschied zwischen Tumor und Peritumorgewebe beobachte t. Eine deutliche lineare Korrelation zwischen der hMTH 1 und der hOGG 1 Exp ression wurde sowohl in Tumor ( r =0.809, P < 0.001) als auch in Peritum orgewebe ( r =0.883, P <0.001) gefunden. Diese Daten sprechen fur ein e reaktive und gegen eine pathogenetische Rolle der untersuchten DNA Reparature nzyme in der Hepatokarzinogenese.
