AIM:To explore the DNA methylation of COL4A1 in ultraviolet-B(UVB)-induced age-related cataract(ARC)models in vitro and in vivo.METHODS:Human lens epithelium B3(HLEB3)cells and Sprague Dawley rats were exposure to UVB...AIM:To explore the DNA methylation of COL4A1 in ultraviolet-B(UVB)-induced age-related cataract(ARC)models in vitro and in vivo.METHODS:Human lens epithelium B3(HLEB3)cells and Sprague Dawley rats were exposure to UVB respectively.The MTT assay was utilized to evaluate cell proliferation.Flow cytometry was employed for analysis of cell apoptosis and cell cycle.COL4A1 expression in HLEB3 cells and anterior lens capsules were assessed using Western blot and reverse transcription-polymerase chain reaction(RTPCR).The localization of COL4A1 in HLEB3 cells was determined by immunofluorescence.The methylation status of CpG islands located in COL4A1 promoter was verified using bisulfite-sequencing PCR(BSP).DNMTs and TETs mRNA levels was examined by RT-PCR.RESULTS:UVB exposure decreased HLEB3 cells proliferation,while increased the apoptosis rate and cells were arrested in G0/G1 phase.COL4A1 expression was markedly inhibited in UVB treated cells compared to the controls.Hypermethylation status was detected in the CpG islands within COL4A1 promoter in HLEB3 cells subjected to UVB exposure.Expressions of DNMTs including DNMT1/2/3 were elevated in UVB treated HLEB3 cells compared to that in the controls,while expressions of TETs including TET1/2/3 showed the opposite trend.Results from the UVB treated rat model further confirmed the decreased expression of COL4A1,hypermethylation status of the CpG islands at promoter of COL4A1 and abnormal expression of DNMT1/2/3 and TET1/2/in UVB exposure group.CONCLUSION:DNA hypermethylation of COL4A1 promoter CpG islands is correlated with decreased COL4A1 expression in UVB induced HLEB3 cells and anterior lens capsules of rats.展开更多
Objectives:Thyroid cancer(THCA)is the most common malignant tumor in endocrine system and the incidence has been increasing worldwide.And the number of patients dying from THCA has also gradually risen because the inc...Objectives:Thyroid cancer(THCA)is the most common malignant tumor in endocrine system and the incidence has been increasing worldwide.And the number of patients dying from THCA has also gradually risen because the incidence continues to increase,so the mechanisms related to effective targets is necessary to improve the survival.This study was to preliminarily investigate the effects of the COL4A2 gene on the regulation of thyroid cancer(THCA)cell proliferation and the associated pathways.Methods:Bioinformatics analysis revealed that COL4A2 was closely associated with cancer development.COL4A2 expression in THCA tissues was analyzed using immunohistochemistry,and survival information was determined via Kaplan-Meier curves.The expression of COL4A2 and AKT pathway-related genes were analyzed using qPCR and western blot analyses.Colony formation as well as CCK-8 assays exhibited the cell proliferation level and cell activity,respectively.Downstream of COL4A2 was identified by Gene set enrichment analysis(GSEA).The effects of the COL4A2 and AKT pathways on THCA tumor growth in vivo were determined using a mouse model.Results:Bioinformatics analysis exhibited that COL4A2 plays a significant role in cancer and that the AKT pathway is downstream of COL4A2.THCA patients with high COL4A2 expression had shorter recurrence-free survival.Upregulation of COL4A2 gene expression in 2 THCA cell lines promoted tumor cell growth and activity.The use of AKT pathway blockers also restrained the growth and activity of the 2 THCA cell lines.The use of AKT pathway blockers reduced tumor volume and mass and prolonged mouse survival.Conclusions:COL4A2 can promote the growth as well as development of THCA through the AKT pathway and COL4A2 could be used as a target for THCA.展开更多
AIM:To evaluate the regulation of the aberrant expression of collagen typeⅣalpha 1 chain(COL4A1)in the development of age-related cataract(ARC).METHODS:Quantitative reverse transcription-polymerase chain reaction(qRT...AIM:To evaluate the regulation of the aberrant expression of collagen typeⅣalpha 1 chain(COL4A1)in the development of age-related cataract(ARC).METHODS:Quantitative reverse transcription-polymerase chain reaction(qRT-PCR)and Western blot analysis were employed to evaluate the expression of COL4A1 in ARC patients and healthy controls.The proliferation,apoptosis,cell cycle and epithelial-mesenchymal transition(EMT)of human lens epithelial cell(HLE-B3)were further analyzed under the condition of COL4A1 gene silence.Alteration of gene expression at mRNA level after knockdown COL4A1 were also evaluated by qRT-PCR on HLE-B3 cells.RESULTS:The aberrant expression of COL4A1 was identified a clinically associated with the ARC.Silencing of COL4A1 promoted the apoptosis and inhibited the proliferation of HLE-B3 by blocking the cell cycle.Moreover,COL4A1 gene silence didn’t affect the cytoskeleton of HLE-B3 but down-regulated the Collagen typeⅣAlpha 2 Chain(COL4A2),paired box 6(PAX6),procollagen-lysine 2-oxoglutarate 5-dioxygenases 1(PLOD1)and procollagenlysine 2-oxoglutarate 5-dioxygenases 2(PLOD2)expression levels in HLE-B3 cells.Silencing the COL4A1 gene induced EMT of the HLE-B3 cells by promoting the transforming growth factor beta(TGF-β)expression.CONCLUSION:Silencing of COL4A1 induces S-phase arrest,also inhibits the proliferation and enhance HLE-B3 apoptosis and EMT,and down-regulates the expression of COL4A2,PAX6,PLOD1 and PLOD2.Thus,the expression alteration of COL4A1 may play a critical role in the pathogenesis of ARC.展开更多
Efficient and stable expression of foreign genes in cells and transgenic animals is important for gain-of-function studies and the establishment of bioreactors.Safe harbor loci in the animal genome enable consistent o...Efficient and stable expression of foreign genes in cells and transgenic animals is important for gain-of-function studies and the establishment of bioreactors.Safe harbor loci in the animal genome enable consistent overexpression of foreign genes,without side effects.However,relatively few safe harbor loci are available in pigs,a fact which has impeded the development of multi-transgenic pig research.We report a strategy for efficient transgene knock-in in the endogenous collagen type I alpha 1 chain(COL1A1)gene using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9(CRISPR/Cas9)system.After the knock-in of a 2A peptide-green fluorescence protein(2A-GFP)transgene in the last codon of COL1A1 in multiple porcine cells,including porcine kidney epithelial(PK15),porcine embryonic fibroblast(PEF)and porcine intestinal epithelial(IPI-2I)cells,quantitative PCR(qPCR),Western blotting,RNA-seq and CCK8 assay were performed to assess the safety of COL1A1 locus.The qPCR results showed that the GFP knock-in had no effect(P=0.29,P=0.66 and P=0.20 for PK15,PEF and IPI-2I cells,respectively)on the mRNA expression of COL1A1 gene.Similarly,no significant differences(P=0.64,P=0.48 and P=0.80 for PK15,PEF and IPI-2I cells,respectively)were found between the GFP knock-in and wild type cells by Western blotting.RNA-seq results revealed that the transcriptome of GFP knock-in PEF cells had a significant positive correlation(P<2.2e–16)with that of the wild type cells,indicating that the GFP knock-in did not alter the global expression of endogenous genes.Furthermore,the CCK8 assay showed that the GFP knock-in events had no adverse effects(P_(24)h=0.31,P_(48)h=0.96,P_(72)h=0.24,P_(96)h=0.17,and P_(120)h=0.38)on cell proliferation of PK15 cells.These results indicate that the COL1A1 locus can be used as a safe harbor for foreign genes knock-in into the pig genome and can be broadly applied to farm animal breeding and biomedical model establishment.展开更多
目的对一个常染色体显性遗传的Van der Hoeve综合征家系进行详尽的临床表型分析及基因突变检测,明确该家系的致病基因突变位点及该突变对基因编码的影响。方法对收集到的Van der Hoeve综合征家系进行包括病史、体格检查及辅助检查在内...目的对一个常染色体显性遗传的Van der Hoeve综合征家系进行详尽的临床表型分析及基因突变检测,明确该家系的致病基因突变位点及该突变对基因编码的影响。方法对收集到的Van der Hoeve综合征家系进行包括病史、体格检查及辅助检查在内的临床资料的收集及外周血液样本的采集,并对22位家系成员进行外显子组测序以及Sanger测序,利用生物信息学软件分析数据。结果该家系共五代,各代连续发病,且每一代男女均可患病,符合常染色体显性遗传特点。该家系中12例患者均自出生时巩膜即呈蓝色且身材矮小,8例患者有骨折病史,可正常愈合,3例患者考虑有Van der Hoeve综合征所致的听力下降,12例患者的COL1A1基因第17号外显子有一个碱基的缺失(c.1128delT),使第376位后的氨基酸编码改变,在第539位提前结束氨基酸编码,该家系中10例无症状者无此突变。结论该家系患者确定为由COL1A1基因c.1128delT突变导致的Van der Hoeve综合征。展开更多
目的探究COL1A2的表达与肺腺癌患者临床病理特征和预后的关系。方法从癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中下载TCGA-肺腺癌的RNA-Seq表达谱和相应的临床数据,通过生物信息学分析COL1A2在肺腺癌组织与正常肺组织中的...目的探究COL1A2的表达与肺腺癌患者临床病理特征和预后的关系。方法从癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中下载TCGA-肺腺癌的RNA-Seq表达谱和相应的临床数据,通过生物信息学分析COL1A2在肺腺癌组织与正常肺组织中的表达差异及与肺腺癌患者生存率的关系。收集2018年1月~2020年12月石河子大学第一附属医院收治的肺腺癌患者82例,采用免疫组化法验证COL1A2的表达与肺腺癌患者临床病理特征及预后的关系。结果COL1A2在肺腺癌组织中呈高表达(P<0.05)。免疫组化结果表明,COL1A2表达与肺腺癌患者TNM分期及是否有远处转移有关(P<0.05);Kaplan-Meier生存分析结果表明,COL1A2的表达与肺腺癌患者预后相关(χ^(2)=9.639,P=0.002);多因素COX回归分析结果表明,COL1A2高表达是肺腺癌患者预后的独立危险因素(HR=2.657,95%CI:1.062~6.646,P=0.037)。结论COL1A2在肺腺癌中呈高表达,与肺腺癌患者肿瘤分期、是否有远处转移及预后相关。展开更多
Propylea japonica(Coleoptera:Coccinellidae)is a natural enemy insect with a wide range of predation in Chinese mainland and is commonly used in pest management.However,its genetic pattern(i.e.,genetic variation,geneti...Propylea japonica(Coleoptera:Coccinellidae)is a natural enemy insect with a wide range of predation in Chinese mainland and is commonly used in pest management.However,its genetic pattern(i.e.,genetic variation,genetic structure,and historical population dynamics)is still unclear,impeding the development of biological control of insect pests.Population genetic research has the potential to optimize strategies at different stages of the biological control processes.This study used 23 nuclear microsatellite sites and mitochondrial COI genes to investigate the population genetics of Propylea japonica based on 462 specimens collected from 30 sampling sites in China.The microsatellite dataset showed a moderate level of genetic diversity,but the mitochondrial genes showed a high level of genetic diversity.Populations from the Yellow River basin were more genetically diverse than those in the Yangtze River basin.Propylea japonica has not yet formed a significant genealogical structure in China,but there was a population structure signal to some extent,which may be caused by frequent gene flow between populations.The species has experienced population expansion after a bottleneck,potentially thanks to the tri-trophic plant–insect–natural enemy relationship.Knowledge of population genetics is of importance in using predators to control pests.Our study complements existing knowledge of an important natural predator in agroecosystems through estimating its genetic diversity and population differentiation and speculating about historical dynamics.展开更多
BACKGROUND Gastric cancer(GC)is a malignant tumor with high morbidity and mortality.Expression of COL5A2 is significantly elevated in GC.Abnormal expression of noncoding RNAs(ncRNAs)have been found in GC,including mic...BACKGROUND Gastric cancer(GC)is a malignant tumor with high morbidity and mortality.Expression of COL5A2 is significantly elevated in GC.Abnormal expression of noncoding RNAs(ncRNAs)have been found in GC,including microRNA(miRNA)and long noncoding RNA(lncRNA).Competing endogenous RNA network plays an important regulatory role in GC.However,its specific regulatory mechanism has not been elucidated.AIM To gain insight into the ncRNA regulatory mechanism and immune microenvironment related to COL5A2 in GC.METHODS RNA sequencing data and clinical information from The Cancer Genome Atlas data portal were used to analyze the expressions of COL5A2,miRNA and lncRNA related to the prognosis of GC.Cox regression analysis and Kyoto Encyclopedia of Genes and Genomes analysis were performed to assess the risk factors and relevant function of COL5A2.StarBase was used to predict the interaction of miRNA–lncRNA or miRNA–mRNA in GC.The relationship between COL5A2,miR-144-3p and ENTPD1-AS1 were verified by dual luciferase reporter assay.The association of COL5A2 with immune cell infiltration were analyzed using the Tumor Immune Estimation Resource database and single sample gene set enrichment analysis.The expression of COL5A2 and macrophages in paired GC tissues were detected by immunohistochemical staining.RESULTS We verified that the upregulation of COL5A2 expression was associated with the prognosis of GC and was an independent risk factor for GC.miR-144-3p was downregulated and correlated with the prognosis of GC.miR-144-3p regulated the expression of COL5A2 through direct interaction with COL5A2.ENTPD1-AS1 was elevated in GC and competitively bound to miR-144-3p,thus inhibiting the expression of miR-144-3p.ENTPD1-AS1 enhanced the expression of COL5A2 through sponging miR-144-3p.Compared to paired normal tissue,COL5A2 expression was upregulated at the protein level,especially in the middle and late stages of GC.The high expression of COL5A2 was positively linked to macrophage infiltration in GC.CONCLUSION COL5A2 regulated by ENTPD1-AS1–miR-144-3p was associated with poor prognosis and macrophage infiltration in GC.This could be a new biomarker and therapeutic target in GC.展开更多
基金Supported by the Shaanxi Provincial Department of Science and Technology Agency Project(No.2022SF-502)Xi’an Medical University Scientific Research Capacity Improvement Project(No.2022NLTS104)+2 种基金The Fifth Batch of Key Disciplines of Xi’an Medical University(No.medical technology12202306)Yunzhen Optometry Special Fund(No.2021HXZR10)Innovation and Entrepreneurship Training Program for College Students(No.S202211840043).
文摘AIM:To explore the DNA methylation of COL4A1 in ultraviolet-B(UVB)-induced age-related cataract(ARC)models in vitro and in vivo.METHODS:Human lens epithelium B3(HLEB3)cells and Sprague Dawley rats were exposure to UVB respectively.The MTT assay was utilized to evaluate cell proliferation.Flow cytometry was employed for analysis of cell apoptosis and cell cycle.COL4A1 expression in HLEB3 cells and anterior lens capsules were assessed using Western blot and reverse transcription-polymerase chain reaction(RTPCR).The localization of COL4A1 in HLEB3 cells was determined by immunofluorescence.The methylation status of CpG islands located in COL4A1 promoter was verified using bisulfite-sequencing PCR(BSP).DNMTs and TETs mRNA levels was examined by RT-PCR.RESULTS:UVB exposure decreased HLEB3 cells proliferation,while increased the apoptosis rate and cells were arrested in G0/G1 phase.COL4A1 expression was markedly inhibited in UVB treated cells compared to the controls.Hypermethylation status was detected in the CpG islands within COL4A1 promoter in HLEB3 cells subjected to UVB exposure.Expressions of DNMTs including DNMT1/2/3 were elevated in UVB treated HLEB3 cells compared to that in the controls,while expressions of TETs including TET1/2/3 showed the opposite trend.Results from the UVB treated rat model further confirmed the decreased expression of COL4A1,hypermethylation status of the CpG islands at promoter of COL4A1 and abnormal expression of DNMT1/2/3 and TET1/2/in UVB exposure group.CONCLUSION:DNA hypermethylation of COL4A1 promoter CpG islands is correlated with decreased COL4A1 expression in UVB induced HLEB3 cells and anterior lens capsules of rats.
文摘Objectives:Thyroid cancer(THCA)is the most common malignant tumor in endocrine system and the incidence has been increasing worldwide.And the number of patients dying from THCA has also gradually risen because the incidence continues to increase,so the mechanisms related to effective targets is necessary to improve the survival.This study was to preliminarily investigate the effects of the COL4A2 gene on the regulation of thyroid cancer(THCA)cell proliferation and the associated pathways.Methods:Bioinformatics analysis revealed that COL4A2 was closely associated with cancer development.COL4A2 expression in THCA tissues was analyzed using immunohistochemistry,and survival information was determined via Kaplan-Meier curves.The expression of COL4A2 and AKT pathway-related genes were analyzed using qPCR and western blot analyses.Colony formation as well as CCK-8 assays exhibited the cell proliferation level and cell activity,respectively.Downstream of COL4A2 was identified by Gene set enrichment analysis(GSEA).The effects of the COL4A2 and AKT pathways on THCA tumor growth in vivo were determined using a mouse model.Results:Bioinformatics analysis exhibited that COL4A2 plays a significant role in cancer and that the AKT pathway is downstream of COL4A2.THCA patients with high COL4A2 expression had shorter recurrence-free survival.Upregulation of COL4A2 gene expression in 2 THCA cell lines promoted tumor cell growth and activity.The use of AKT pathway blockers also restrained the growth and activity of the 2 THCA cell lines.The use of AKT pathway blockers reduced tumor volume and mass and prolonged mouse survival.Conclusions:COL4A2 can promote the growth as well as development of THCA through the AKT pathway and COL4A2 could be used as a target for THCA.
基金Supported by Supporting Fund Project of Shaanxi Provincial Department of Science and Technology Agency Project(No.2022SF-502)Special Scientific Research Program of Education Department of Shaanxi Provincial Government(No.21JK0891)+1 种基金Young Talent Lifting Project of Xi’an Science and Technology Association(No.095920221365)Innovation and Entrepreneurship Training Program for College students of Xi’an Medical University(No.121521113)。
文摘AIM:To evaluate the regulation of the aberrant expression of collagen typeⅣalpha 1 chain(COL4A1)in the development of age-related cataract(ARC).METHODS:Quantitative reverse transcription-polymerase chain reaction(qRT-PCR)and Western blot analysis were employed to evaluate the expression of COL4A1 in ARC patients and healthy controls.The proliferation,apoptosis,cell cycle and epithelial-mesenchymal transition(EMT)of human lens epithelial cell(HLE-B3)were further analyzed under the condition of COL4A1 gene silence.Alteration of gene expression at mRNA level after knockdown COL4A1 were also evaluated by qRT-PCR on HLE-B3 cells.RESULTS:The aberrant expression of COL4A1 was identified a clinically associated with the ARC.Silencing of COL4A1 promoted the apoptosis and inhibited the proliferation of HLE-B3 by blocking the cell cycle.Moreover,COL4A1 gene silence didn’t affect the cytoskeleton of HLE-B3 but down-regulated the Collagen typeⅣAlpha 2 Chain(COL4A2),paired box 6(PAX6),procollagen-lysine 2-oxoglutarate 5-dioxygenases 1(PLOD1)and procollagenlysine 2-oxoglutarate 5-dioxygenases 2(PLOD2)expression levels in HLE-B3 cells.Silencing the COL4A1 gene induced EMT of the HLE-B3 cells by promoting the transforming growth factor beta(TGF-β)expression.CONCLUSION:Silencing of COL4A1 induces S-phase arrest,also inhibits the proliferation and enhance HLE-B3 apoptosis and EMT,and down-regulates the expression of COL4A2,PAX6,PLOD1 and PLOD2.Thus,the expression alteration of COL4A1 may play a critical role in the pathogenesis of ARC.
基金supported by the Major Scientific Research Tasks for Scientific and Technological Innovation Projects of the Chinese Academy of Agricultural Sciences(CAAS-ZDRW202006)the National Transgenic Breeding Project(2018ZX08010-10B)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences(ASTIP-IAS05).
文摘Efficient and stable expression of foreign genes in cells and transgenic animals is important for gain-of-function studies and the establishment of bioreactors.Safe harbor loci in the animal genome enable consistent overexpression of foreign genes,without side effects.However,relatively few safe harbor loci are available in pigs,a fact which has impeded the development of multi-transgenic pig research.We report a strategy for efficient transgene knock-in in the endogenous collagen type I alpha 1 chain(COL1A1)gene using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9(CRISPR/Cas9)system.After the knock-in of a 2A peptide-green fluorescence protein(2A-GFP)transgene in the last codon of COL1A1 in multiple porcine cells,including porcine kidney epithelial(PK15),porcine embryonic fibroblast(PEF)and porcine intestinal epithelial(IPI-2I)cells,quantitative PCR(qPCR),Western blotting,RNA-seq and CCK8 assay were performed to assess the safety of COL1A1 locus.The qPCR results showed that the GFP knock-in had no effect(P=0.29,P=0.66 and P=0.20 for PK15,PEF and IPI-2I cells,respectively)on the mRNA expression of COL1A1 gene.Similarly,no significant differences(P=0.64,P=0.48 and P=0.80 for PK15,PEF and IPI-2I cells,respectively)were found between the GFP knock-in and wild type cells by Western blotting.RNA-seq results revealed that the transcriptome of GFP knock-in PEF cells had a significant positive correlation(P<2.2e–16)with that of the wild type cells,indicating that the GFP knock-in did not alter the global expression of endogenous genes.Furthermore,the CCK8 assay showed that the GFP knock-in events had no adverse effects(P_(24)h=0.31,P_(48)h=0.96,P_(72)h=0.24,P_(96)h=0.17,and P_(120)h=0.38)on cell proliferation of PK15 cells.These results indicate that the COL1A1 locus can be used as a safe harbor for foreign genes knock-in into the pig genome and can be broadly applied to farm animal breeding and biomedical model establishment.
文摘目的对一个常染色体显性遗传的Van der Hoeve综合征家系进行详尽的临床表型分析及基因突变检测,明确该家系的致病基因突变位点及该突变对基因编码的影响。方法对收集到的Van der Hoeve综合征家系进行包括病史、体格检查及辅助检查在内的临床资料的收集及外周血液样本的采集,并对22位家系成员进行外显子组测序以及Sanger测序,利用生物信息学软件分析数据。结果该家系共五代,各代连续发病,且每一代男女均可患病,符合常染色体显性遗传特点。该家系中12例患者均自出生时巩膜即呈蓝色且身材矮小,8例患者有骨折病史,可正常愈合,3例患者考虑有Van der Hoeve综合征所致的听力下降,12例患者的COL1A1基因第17号外显子有一个碱基的缺失(c.1128delT),使第376位后的氨基酸编码改变,在第539位提前结束氨基酸编码,该家系中10例无症状者无此突变。结论该家系患者确定为由COL1A1基因c.1128delT突变导致的Van der Hoeve综合征。
文摘目的探究COL1A2的表达与肺腺癌患者临床病理特征和预后的关系。方法从癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中下载TCGA-肺腺癌的RNA-Seq表达谱和相应的临床数据,通过生物信息学分析COL1A2在肺腺癌组织与正常肺组织中的表达差异及与肺腺癌患者生存率的关系。收集2018年1月~2020年12月石河子大学第一附属医院收治的肺腺癌患者82例,采用免疫组化法验证COL1A2的表达与肺腺癌患者临床病理特征及预后的关系。结果COL1A2在肺腺癌组织中呈高表达(P<0.05)。免疫组化结果表明,COL1A2表达与肺腺癌患者TNM分期及是否有远处转移有关(P<0.05);Kaplan-Meier生存分析结果表明,COL1A2的表达与肺腺癌患者预后相关(χ^(2)=9.639,P=0.002);多因素COX回归分析结果表明,COL1A2高表达是肺腺癌患者预后的独立危险因素(HR=2.657,95%CI:1.062~6.646,P=0.037)。结论COL1A2在肺腺癌中呈高表达,与肺腺癌患者肿瘤分期、是否有远处转移及预后相关。
基金supported by a grant from the Key S&T Special Project of Henan,China(201300111500)the National Key R&D Program of China(2018YFD0200600)+1 种基金the Modern Agricultural System in Industry Technology of Henan Province,China(S2015-02-G05)the Key R&D and Promotion Project in Henan Province,China(212102110471).
文摘Propylea japonica(Coleoptera:Coccinellidae)is a natural enemy insect with a wide range of predation in Chinese mainland and is commonly used in pest management.However,its genetic pattern(i.e.,genetic variation,genetic structure,and historical population dynamics)is still unclear,impeding the development of biological control of insect pests.Population genetic research has the potential to optimize strategies at different stages of the biological control processes.This study used 23 nuclear microsatellite sites and mitochondrial COI genes to investigate the population genetics of Propylea japonica based on 462 specimens collected from 30 sampling sites in China.The microsatellite dataset showed a moderate level of genetic diversity,but the mitochondrial genes showed a high level of genetic diversity.Populations from the Yellow River basin were more genetically diverse than those in the Yangtze River basin.Propylea japonica has not yet formed a significant genealogical structure in China,but there was a population structure signal to some extent,which may be caused by frequent gene flow between populations.The species has experienced population expansion after a bottleneck,potentially thanks to the tri-trophic plant–insect–natural enemy relationship.Knowledge of population genetics is of importance in using predators to control pests.Our study complements existing knowledge of an important natural predator in agroecosystems through estimating its genetic diversity and population differentiation and speculating about historical dynamics.
基金Supported by National Natural Science Foundation of China,No.81971489Natural Science Foundation of Guangdong Province,No.2022A1515011122。
文摘BACKGROUND Gastric cancer(GC)is a malignant tumor with high morbidity and mortality.Expression of COL5A2 is significantly elevated in GC.Abnormal expression of noncoding RNAs(ncRNAs)have been found in GC,including microRNA(miRNA)and long noncoding RNA(lncRNA).Competing endogenous RNA network plays an important regulatory role in GC.However,its specific regulatory mechanism has not been elucidated.AIM To gain insight into the ncRNA regulatory mechanism and immune microenvironment related to COL5A2 in GC.METHODS RNA sequencing data and clinical information from The Cancer Genome Atlas data portal were used to analyze the expressions of COL5A2,miRNA and lncRNA related to the prognosis of GC.Cox regression analysis and Kyoto Encyclopedia of Genes and Genomes analysis were performed to assess the risk factors and relevant function of COL5A2.StarBase was used to predict the interaction of miRNA–lncRNA or miRNA–mRNA in GC.The relationship between COL5A2,miR-144-3p and ENTPD1-AS1 were verified by dual luciferase reporter assay.The association of COL5A2 with immune cell infiltration were analyzed using the Tumor Immune Estimation Resource database and single sample gene set enrichment analysis.The expression of COL5A2 and macrophages in paired GC tissues were detected by immunohistochemical staining.RESULTS We verified that the upregulation of COL5A2 expression was associated with the prognosis of GC and was an independent risk factor for GC.miR-144-3p was downregulated and correlated with the prognosis of GC.miR-144-3p regulated the expression of COL5A2 through direct interaction with COL5A2.ENTPD1-AS1 was elevated in GC and competitively bound to miR-144-3p,thus inhibiting the expression of miR-144-3p.ENTPD1-AS1 enhanced the expression of COL5A2 through sponging miR-144-3p.Compared to paired normal tissue,COL5A2 expression was upregulated at the protein level,especially in the middle and late stages of GC.The high expression of COL5A2 was positively linked to macrophage infiltration in GC.CONCLUSION COL5A2 regulated by ENTPD1-AS1–miR-144-3p was associated with poor prognosis and macrophage infiltration in GC.This could be a new biomarker and therapeutic target in GC.
