Quantitative trait loci identification reveals zinc finger protein CONSTANS-LIKE 4 as the key candidate gene of stigma color in watermelon(Citrullus lanatus)

Stigma color is a critical agronomic trait in watermelon that plays an important role in pollination.However,there are few reports on the regulation of stigma color in watermelon.In this study,a genetic analysis of the F2 population derived from ZXG1553(P1,with orange stigma)and W1-17(P2,with yellow stigma)indicated that stigma color is a quantitative trait and the orange stigma is recessive compared with the yellow stigma.Bulk segregant analysis sequencing(BSA-seq)revealed a 3.75 Mb segment on chromosome 6 that is related to stigma color.Also,a major stable effective QTL Clqsc6.1(QTL stigma color)was detected in two years between cleaved amplified polymorphic sequencing(CAPS)markers Chr06_8338913 and Chr06_9344593 spanning a~1.01 Mb interval that harbors 51 annotated genes.Cla97C06G117020(annotated as zinc finger protein CONSTANS-LIKE 4)was identified as the best candidate gene for the stigma color trait through RNA-seq,quantitative real-time PCR(qRT-PCR),and gene structure alignment analysis among the natural watermelon panel.The expression level of Cla97C06G117020 in the orange stigma accession was lower than in the yellow stigma accessions with a significant difference.A nonsynonymous SNP site of the Cla97C06G117020 coding region that causes amino acid variation was related to the stigma color variation among nine watermelon accessions according to their re-sequencing data.Stigma color formation is often related to carotenoids,and we also found that the expression trend of ClCHYB(annotated asβ-carotene hydroxylase)in the carotenoid metabolic pathway was consistent with Cla97C06G117020,and it was expressed in low amounts in the orange stigma accession.These data indicated that Cla97C06G117020 and ClCHYB may interact to form the stigma color.This study provides a theoretical basis for gene fine mapping and mechanisms for the regulation of stigma color in watermelon.

Expression Pattern Analysis of Zinc Finger Protein Genes in Wheat(Triticum aestivum L.) Under Phosphorus Deprivation

Zinc finger protein（ZFP） genes comprise a large and diverse gene family, and are involved in biotic and abiotic stress responses in plants. In this study, a total of 126 ZFP genes classified into various types in wheat were characterized and subjected to expression pattern analysis under inorganic phosphate（Pi） deprivation. The wheat ZFP genes and their corresponding GenBank numbers were obtained from the information of a 4×44K wheat gene expression microarray chip. They were confirmed by sequence similarity analysis and named based on their homologs in Brachypodium distachyon or Oriza sativa. Expression analysis based on the microarray chip revealed that these ZFP genes are categorized into 11 classes according to their gene expression patterns in a 24-h of Pi deprivation regime. Among them, ten genes were differentially up-regulated, ten genes differentially downregulated, and two genes both differentially up- and down-regulated by Pi deprivation. The differentially up- or down-regulated genes exhibited significantly more or less transcripts at one, two, or all of the checking time points（1, 6, and 24 h） of Pi stress in comparison with those of normal growth, respectively. The both differentially up- and down-regulated genes exhibited contrasting expression patterns, of these, TaWRKY70;5 showed significantly up-regulated at 1 and 6 h and down-regulated at 24 h whereas TaAN1AN20-8;2 displayed significantly upregulated at 1 h and downregulated at 6 h under deprivation Pi condition. Real time PCR analysis confirmed the expression patterns of the differentially expressed genes obtained by the microarray chip. Our results indicate that numerous ZFP genes in wheat respond to Pi deprivation and have provided further insight into the molecular basis that plants respond to Pi deprivation mediated by the ZFP gene family.

Molecular Cloning and Expression Analysis of a Cys2/His2 Type Zinc Finger Protein Gene in Upland Cotton

The zinc finger proteins belong to the largest family of transcription factors.But there is little research of Cys2/His2 type zinc finger proteins in cotton,and there is no submission of correlating

Zinc finger protein A20 protects rats against chronic liver allograft dysfunction

AIM:To investigate the effect of zinc finger protein A20 on chronic liver allograft dysfunction in rats.METHODS:Allogeneic liver transplantation from DA rats to Lewis rats was performed.Chronic liver allograft dysfunction was induced in the rats by administering low-dose tacrolimus at postoperative day (POD) 5.Hepatic overexpression of A20 was achieved by recombinant adenovirus (rAd.)-mediated gene transfer administered intravenously every 10 d starting from POD 10.The recipient rats were injected with physiological saline,rAdEasy-A20 (1 × 109 pfu/30 g weight) or rAdEasy (1 × 109 pfu/30 g weight) every 10 d through the tail vein for 3 mo starting from POD 10.Liver tissue samples were harvested on POD 30 and POD 60.RESULTS:Liver-transplanted rats treated with only tacrolimus showed chronic allograft dysfunction with severe hepatic fibrosis.A20 overexpression ameliorated the effects on liver function,attenuated liver allograft fibrosis and prolonged the survival of the recipient rats.Treatment with A20 suppressed hepatic protein production of tumor growth factor (TGF)1,interleukin1 ,caspase-8,CD40,CD40L,intercellular adhesion molecule-1,vascular cell adhesion molecule-1 and E-selectin.A20 treatment suppressed liver cell apoptosis and inhibited nuclear factorB activation of Kupffer cells (KCs),liver sinusoidal endothelial cells (LSECs) and hepatic stellate cells (HSCs),and it subsequently decreased cytokine mRNA expression in KCs and LSECs and reduced the production of TGF1 in HSCs.CONCLUSION:A20 might prevent chronic liver allograft dysfunction by re-establishing functional homeostasis of KCs,LSECs and HSCs.

Zinc Finger Protein-activating Transcription Factor Up-regulates Vascular Endothelial Growth Factor-A Expression in Vitro

Objective To construct the zinc finger protein-activating transcription factor (ZFP-ATF) plasmid and evaluate its efficacy in inducing vascular endothelial growth factor (VEGF) expression in EY.HY926 endothelial cells. Methods Firstly, we constructed the ZFP-ATF plasmid, then testified the quantity of VEGF protein in EY.HY926 endothelial cells after transfected with ZFP-ATP plasmid by Western blot, finally, we used the RT-PCR to testify whether the ZFP-ATF can stimulate expression of VEGF splice variants. Results The ZFP-ATF DNA sequences were located the multiclone sites of PVAX1 vector between the site of BamH Ⅰ and Xhol. Western blot result showed VEGF expression in EY.HY926 endothelial cells transfected with ZFP-ATF plasmid was significantly higher than that in cells transfected with VEGF165 (19.95±3.95 vs. 12.15±1.55 μg/μL, P<0.01). RT-PCR result showed VEGF-A mRNA expression level induced by ZFP-ATF was high than that induced by VEGF165. Conclusion ZFP-ATF can up-regulate the VEGF-A expression in comparison with VEGF165, which might have beneficial effects in angiogenesis process.

Zinc finger protein 139 expression in gastric cancer and its clinical significance

AIM:To investigate the expression of zinc finger protein 139(ZNF139)in gastric cancer(GC),and to analyze its clinical significance.METHODS:A total of 108 patients who were diagnosed with GC and underwent surgery between January 2005and March 2007 were enrolled in this study.Gastric tumor specimens and paired tumor-adjacent tissues were collected and paraffin-embedded,and the clinicopathologic characteristics and prognosis were recorded.The expression of ZNF139,Bcl-2,Bax,and caspase-3 were determined by immunohistochemistry,and apoptosis was assessed by terminal deoxynucleotidyl transferasemediated d UTP-biotin nick end labeling.SPSS 13.0software was used for data processing and analyses,and significance was determined at P<0.05.RESULTS:The expression of ZNF139 was stronger in tumors than in tumor-adjacent tissues(66.67%vs 44.44%;P<0.01).Overexpression of ZNF139correlated with tumor differentiation,invasion depth,clinical stage,lymphatic metastasis,and blood vessel invasion(all P s<0.05).Patients with overexpression of ZNF139 had a poorer prognosis(P<0.01),and overexpression of ZNF139 was an independent factor for the prognosis of GC patients by a Cox survival analysis(P=0.02).A negative relationship between ZNF139 and the apoptosis index was observed(r=-0.686;P<0.01).The expression of Bcl-2 in GC was stronger than in tumor-adjacent tissues(66.67%vs41.67%),whereas the expression levels of Bax and caspase-3 were lower in primary tumors(54.63%and47.22%,respectively)than in tumor-adjacent tissues(73.15%and 73.15%,respectively)(all Ps<0.05).The expression of ZNF139 negatively correlated with caspase-3(r=-0.370;P<0.01).The expressions of Bcl-2 and Bax were also negatively correlated(r=-0.231;P=0.02).The expressions of caspase-3 and Bax protein were positively correlated(r=0.217;P=0.024).CONCLUSION:ZNF139 is related to clinicopathologic characteristics and prognosis of GC.Furthermore,it is overexpressed and involved in apoptosis in GC tissues by regulating caspase-3.

The C_2H_2 -type Zinc Finger Protein ZFP182 is Involved in Abscisic Acid-Induced Antioxidant Defense in Rice

C2H2-type zinc finger proteins （ZFPs） are thought to play important roles in modulating the responses of plants to drought, salinity and oxidative stress. However, direct evidence is lacking for the involvement of these ZFPs in abscisic acid （ABA）-induced antioxidant defense in plants. In this study, the role of the rice （Oryza sativa L. sub.japonica cv. Nipponbare） C2H2-type ZFP ZFP182 in ABA-induced antioxidant defense and the relationship between ZFP182 and two rice MAPKs, OsMPK1 and OsMPK5 in ABA signaling were investigated. ABA treatment induced the increases in the expression of ZFP182, OsMPK1 and OsMPK5, and the activities of superoxide dismutase （SOD） and ascorbate peroxidase （APX） in rice leaves. The transient gene expression analysis and the transient RNA interference （RNAi） analysis in protoplasts showed that ZFP182, OsMPK1 and OsMPK5 are involved in ABA-induced up-regulation in the activities of SOD and APX. Besides, OsMPK1 and OsMPK5 were shown to be required for the up-regulation in the expression of ZFP182 in ABA signaling, but ZFP182 did not mediate the ABA-induced up-regulation in the expression of OsMPK1 and OsMPKS. These results indicate that ZFP182 is required for ABA-induced antioxidant defense and the expression of ZFP182 is regulated by rice MAPKs in ABA signaling.

A zinc finger protein, interacted with cyclophilin,affects root development via IAA pathway in rice

The plant hormone auxin plays a crucial role in lateral root development. To better understand the rnolecular mechanisms underlying lateral root formation, an auxin-responsive gene OsCYP2 （Oso2g0121300） was characterized from rice. Compared to the wild type, OsCYP2-RNAi （RNA interference） lines exhibited distinctive defects in lateral root development. Yeast two-hybrid and glutathione S-transferase puIl-down results confirmed that OsCYP2 interacted with a C2HC-type zinc finger protein （OsZFP, O501g0252900） which is located in the rice nucleus. T2OsZFP-RNAi lines had significantly fewer lateral roots than did wild-type plants, which suggests a role for OsCYP2 and OsZFP in regulating lateral root development.Quantitative real-time polymerase chain reaction showed that the expression of certain Aux/IAA （auxin/indole-3- acetic acid） genes was altered in OsCYP2- and OsZFP-RNAi lines in response to IAA. These findings imply that OsCYP2 and OsZFP participate in IAA signal pathways controlling lateral root development. More importantly, OslAA11 showed functional redundancy not only in OsCYP2-RNAi lines but also in OsZFP-RNAi lines, which provides important clues for the elucidation of mechanisms controlling lateral root development in response to auxin.

Involvement of C2H2 zinc finger proteins in the regulation of epidermal cell fate determination in Arabidopsis

Cell fate determination is a basic developmental process during the growth of multicellular organisms.Trichomes and root hairs of Arabidopsis are both readily accessible structures originating from the epidermal cells of the aerial tissues and roots respectively, and they serve as excellent models for understanding the molecular mechanisms controlling cell fate determination and cell morphogenesis. The regulation of trichome and root hair formationis a complex program that consists of the integration of hormonal signals with a large number of transcriptional factors, including MYB and b HLH transcriptional factors.Studies during recent years have uncovered an important role of C2H2 type zinc finger proteins in the regulation of epidermal cell fate determination. Here in this minireview we briefly summarize the involvement of C2H2 zinc finger proteins in the control of trichome and root hair formation in Arabidopsis.

Fruit size control by a zinc finger protein regulating pericarp cell size in tomato

Fruit size is largely defined by the number and size of cells in the fruit.Endoreduplication–a specialized cell cycle–is highly associated with cell expansion during tomato fruit growth.However,how endoreduplication coupled with cell size is regulated remains poorly understood.In this study,we identified a zinc finger gene SlPZF1(Solanum lycopersicum PERICARP-ASSOCIATED ZINC FINGER PROTEIN 1)that was highly expressed in the pericarp of developing fruits.Plants with altered SlPZF1 expression produced smaller fruits due to the reduction in cell size associated with weakened endoreduplication.Overexpressing SlPZF1 delayed cell division phase by enhancing early expression of several key cell cycle regulators including SlCYCD3;1 and two plant specific mitotic cyclin-dependent protein kinase(SlCDKB1 and SlCDKB2)in the pericarp tissue.Furthermore,we identified 14 putative SlPZF1 interacting proteins(PZFIs)via yeast two hybrid screening.Several PZFIs,including Pre-mRNA-splicing factor(SlSMP1/PZFI4),PAPA-1-like conserved region family protein(PZFI6),Fanconi anemia complex components(PZFI3 and PZFI10)and bHLH transcription factor LONESOME HIGHWAY(SILHW/PZFI14),are putatively involved in cell cycle regulation.Our results demonstrate that fruit growth in tomato requires balanced expression of the novel cell size regulator SlPZF1.

Cloning of a novel human zinc finger protein(ZNF199)cDNA

ZINC finger proteins play an important role in the regulation of gene expression by binding to DNA/RNA in a sequence-specific manner. Now nearly 200 human zinc finger genes have been cloned. According to the difference of the conservative domain, the zinc finger proteins can be classified into several types.Most of zinc finger proteins belong to the C2H2 type containing the consensus amino acid sequence YECX2CX3FX5LX2HX3HTGEKP, which is rather conservative especially in the link region between two zinc finger motifs （TGEKP）

The roles of zinc finger proteins in non-alcoholic fatty liver disease

Non-alcoholic fatty liver disease(NAFLD)is a common chronic disease characterized by excessive fat accumulation in hepatocytes in the absence of alcohol consumption.Modern trends towards excessive calorie intake and sedentary life styles have increased the prevalence of NAFLD accompanied by obesity and type 2 diabetes.However,the molecular mechanisms underlying the initiation and progression of NAFLD are not clear.Zinc finger proteins(ZFPs)are a superfamily of metalloproteins that contain zinc finger motifs.ZFPs play diverse physiological roles in tissue homeostasis and also contribute to many pathological conditions,including metabolic,cardiovascular,and neurodegenerative diseases and various types of cancer.In this review,we highlight our current knowledge of several ZFPs that play critical roles in the progression of NAFLD,describe their mechanistic functional networks,and discuss the potential for ZFPs as therapeutic targets for NAFLD.

Expression of putative zinc-finger protein lcn61 gene in lymphocystis disease virus China (LCDV-cn) genome

An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified and inserted into pET24a (+) vector. Then it expressed in E. coli BL21 (DE3), and His-tag fusion protein of high yield was obtained. It was found that the fusion protein existed in E. coli mainly as inclusion bodies. The bioinformatics analysis indicates that LCN61 is C2H2 type zinc-finger protein containing four C2H2 zinc-finger motifs. This work provides a theory for functional research of lcn61 gene.

Expression and purification of a novel ZNF191 zinc finger protein——ZNF191 protein and its truncated zinc finger region ZNF191 (243—368)

ZNF191 is a new zine finger gene whieh has a 1107 bp open reading frame （ORF） and eneodes a 368 amino avid protein including a putative DNA-binding domain of four Cys2 His2 zine finger motifs at its C-terminal region. The ZNF191 cDNA is loeated in the chromosome 18q12.1 region where it is known that a variety of hereditary diseases are related to. Probably, this protein has potential function of stimulating the gene franscription. In order to examine the function and structure of ZNF191 protein, the ORF of ZNF191 and its zine finger region ZNF191（243--368） genes were inserted into PTSA-18 expression vector by PCR amplification, then the constructed genes were expressed in the PTSA-18/B121 （DE3） system. The two proteins were purified by DEAE-52, CM-23 and Heparin-Sepharose 4B columns. The pooled proteins showed a single band as assayed by Coomasie Brillian Blue Staining of an SDS/polyacryamide gel.

Zinc finger structure-function in Ikaros

The zinc finger motif was used as a vehicle for the initial discovery of Ikaros in the context of T-cell differentiation and has been central to all subsequent analyses of Ikaros function.The Ikaros gene is alternately spliced to produce several isoforms that confer diversity of function and consequently have complicated analysis of the function of Ikaros in vivo.Key features of Ikaros in vivo function are associated with six C2H2 zinc fingers;four of which are alternately incorporated in the production of the various Ikaros isoforms.Although no complete structures are available for the Ikaros protein or any of its family members,considerable evidence has accumulated about the structure of zinc fingers and the role that this structure plays in the functions of the Ikaros family of proteins.This review summarizes the structural aspects of Ikaros zinc fingers,individually,and in tandem to provide a structural context for Ikaros function and to provide a structural basis to inform the design of future experiments with Ikaros and its family members.

斑点叉尾鮰ZBTB38的原核表达、多克隆抗体制备及应用

为获得斑点叉尾鮰(Ictalurus punctatus)ZBTB38的多克隆抗体,并研究其在性腺中的表达情况,将斑点叉尾鮰zbtb38基因部分序列经密码子优化后连接至pET32a(+)载体,构建重组质粒pET32a(+)-zbtb38,再转入大肠杆菌(Escherichia coli)感受态细胞BL21(DE3)中诱导表达,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、免疫印迹(Western blot)和液相色谱-质谱联用(LC-MS)等技术进行鉴定。用纯化后的重组蛋白制备兔抗斑点叉尾鮰ZBTB38多克隆抗体,通过酶联免疫吸附(ELISA)和Western blot技术检测抗体效价及其特异性,最后采用Western blot方法检测ZBTB38在性腺中的表达情况,并使用实时荧光定量PCR(qRT-PCR)技术对检测结果进行验证。结果表明,制备的兔抗斑点叉尾鮰ZBTB38多克隆抗体效价可达1:(5.12×10^(5)),能够特异性识别斑点叉尾鮰性腺组织中表达的ZBTB38蛋白,其中精巢组织中的表达水平显著高于卵巢,Western blot与qRT-PCR检测结果较为一致。综上所述,研究成功制备了斑点叉尾鮰ZBTB38的多克隆抗体,为下一步深入研究斑点叉尾鮰zbtb38基因的功能奠定了基础。

脓毒症并发急性肾损伤患者血清HDAC4和KLF5的表达及其临床价值

目的探究脓毒症并发急性肾损伤(AKI)患者血清组蛋白去乙酰基转移酶4(HDAC4)和锌指蛋白转录因子5(KLF5)表达及临床价值。方法选取2020年9月—2022年9月本院收治的60例脓毒症并发AKI患者作为AKI组,选取同期北京市大兴区人民医院收治的75例脓毒症未发生AKI患者作为非AKI组。比较两组的临床资料、血清HDAC4和KLF5水平。ROC分析血清HDAC4和KLF5对脓毒症患者并发AKI的诊断价值。Logistic回归分析影响脓毒症患者并发AKI的因素。结果与非AKI组相比,AKI组患者血清HDAC4、KLF5、降钙素原(PCT)、肌酐(Cr)、序贯性器官衰竭(SOFA)、急性生理学及慢性健康状况评分系统Ⅱ(APACHEⅡ)评分较高,AKI组血小板计数(PLT)较低,差异有统计学意义(P<0.05)。随着AKI组患者分期升高,血清HDAC4和KLF5水平依次升高,差异有统计学意义(P<0.05)。ROC曲线分析显示,血清HDAC4、KLF5可辅助诊断脓毒症患者是否并发AKI的曲线下面积(AUC)是0.800(95%CI:0.723~0.876)、0.810(95%CI:0.735~0.886);二者联合诊断的AUC为0.908(95%CI:0.856~0.961),均优于各自单独检测(Z=2.277、2.102,P<0.05)。Logistic回归分析显示,APACHEⅡ评分、SOFA评分、HDAC4、KLF5是影响脓毒症患者是否并发AKI的危险因素(P<0.05)。结论脓毒症并发AKI患者血清HDAC4、KLF5水平升高,且二者联合检测对脓毒症并发AKI的诊断效能较高,对评估脓毒症并发AKI有较好的临床诊断价值。

miR-592靶向调控PRDM5影响肾细胞癌侵袭转移及自噬的机制研究

目的探究miR-592靶向调控PR结构域锌指蛋白5(PR domain zinc finger protein 5,PRDM5)影响肾细胞癌(renal cell carcinoma,RCC)侵袭转移及自噬的机制。方法RT-qPCR、蛋白质印迹检测RCC组织和细胞中miR-592、PRDM5 mRNA和蛋白水平。A498、786-O细胞分为anti-miR-NC组、anti-miR-592组、anti-miR-592+si-NC组、anti-miR-592+si-PRDM5组。RNA免疫共沉淀(RIP)检测miR-592和PRDM5的结合;CCK-8、Transwell实验检测细胞增殖、迁移和侵袭能力;透射电镜观察自噬小体形成;蛋白质印迹检测细胞PRDM5、基质金属蛋白酶(MMP)-2、MMP-9、Beclin1、微管相关蛋白1轻链3(LC3)Ⅱ/Ⅰ水平。建立移植瘤裸鼠模型,分析敲低miR-592表达对移植瘤生长的影响。结果RCC组织和细胞中PRDM5 mRNA和蛋白水平降低,miR-592水平增加(P<0.05)。敲低miR-592表达后,细胞A_(450 nm)、迁移和侵袭细胞数、MMP-2、MMP-9水平降低,自噬小体数量、Beclin1、LC3Ⅱ/Ⅰ水平增加(P<0.05)。敲低PRDM5表达能部分逆转敲低miR-592对细胞增殖、侵袭及自噬的影响(P<0.05)。敲低miR-592表达能够抑制裸鼠移植瘤生长(P<0.05)。结论miR-592通过靶向抑制PRDM5表达,促进RCC细胞增殖、侵袭转移能力,并抑制自噬能力。

ZNF772甲基化及HR-HPV检测在意义不明的非典型鳞状细胞病人分流中的意义

目的:探讨锌指蛋白(ZNF)772甲基化及高危型人乳头瘤病毒(HR-HPV)检测在意义不明的非典型鳞状细胞(ASCUS)病人分流中的意义。方法:选取初筛ASCUS病人195例,均接受HC2-HPV-DNA检测及ZNF772基因DNA甲基化检测;比较宫颈上皮内瘤样变(CIN)Ⅱ及以上与CINⅡ以下、CINⅢ及以上及CINⅢ以下病人ZNF772基因位点甲基化水平的差异,并评估ZNF772甲基化水平及HR-HPV对ASCUS病人CINⅡ及以上、CINⅢ及以上的分流价值。结果:病理学检查显示,195例ASCUS病人中炎症80例(41.03%)、CINⅠ64例(32.82%)、CINⅡ29例(14.87%)、CINⅢ/CIS/AIS 18例(9.23%)、SCC/AC 4例(2.05%)。HR-HPV(HC2)阳性141例(72.31%)、HR-HPV(HC2)阴性54例(27.69%);CINⅡ及以上病人HR-HPV(HC2)阳性检出率高于CINⅡ以下(P<0.05);CINⅢ及以上病人HR-HPV(HC2)阳性检出率高于CINⅢ以下(P<0.05)。CINⅡ及以上、CINⅢ及以上病人ZNF772基因-420、-422位点甲基化水平分别高于CINⅡ以下、CINⅢ以下病人(P<0.05)。ZNF772基因-420、-422位点甲基化水平及HR-HPV诊断CINⅡ及以上的AUC(95%CI)分别为0.867(0.811~0.911)、0.806(0.743~0.859)、0.777(0.712~0.834),诊断CINⅢ及以上的AUC(95%CI)分别为0.891(0.838~0.931)、0.868(0.813~0.912)、0.780(0.716~0.836)。ZNF772基因-420位点甲基化水平诊断CINⅡ及以上、CINⅢ及以上的AUC面积最大。结论:相较于HC2-HPV-DNA检测,ZNF772甲基化检测更能准确地识别ASCUS中CINⅡ及以上、CINⅢ及以上病变病人,有望成为分流ASCUS的有效替代手段。

ZNF554 Inhibits Endometrial Cancer Progression via Regulating RBM5 and Inactivating WNT/β-Catenin Signaling Pathway

Objective:Uterine corpus endometrial carcinoma(UCEC),a kind of gynecologic malignancy,poses a significant risk to women’s health.The precise mechanism underlying the development of UCEC remains elusive.Zinc finger protein 554(ZNF554),a member of the Krüppel-associated box domain zinc finger protein superfamily,was reported to be dysregulated in various illnesses,including malignant tumors.This study aimed to examine the involvement of ZNF554 in the development of UCEC.Methods:The expression of ZNF554 in UCEC tissues and cell lines were examined by qRT-PCR and Western blot assay.Cells with stably overexpressed or knocked-down ZNF554 were established through lentivirus infection.CCK-8,wound healing,and Transwell invasion assays were employed to assess cell proliferation,migration,and invasion.Propidium iodide(PI)staining combined with fluorescence-activated cell sorting(FACS)flow cytometer was utilized to detect cell cycle distribution.qRT-PCR and Western blotting were conducted to examine relative mRNA and protein levels.Chromatin immunoprecipitation assay and luciferase reporter assay were used to explore the regulatory role of ZNF554 in RNA binding motif 5(RBM5).Results:The expression of ZNF554 was found to be reduced in both UCEC samples and cell lines.Decreased expression of ZNF554 was associated with higher tumor stage,decreased overall survival,and reduced disease-free survival in UCEC.ZNF554 overexpression suppressed cell proliferation,migration,and invasion,while also inducing cell cycle arrest.In contrast,a decrease in ZNF554 expression resulted in the opposite effect.Mechanistically,ZNF554 transcriptionally regulated RBM5,leading to the deactivation of the Wingless(WNT)/β-catenin signaling pathway.Moreover,the findings from rescue studies demonstrated that the inhibition of RBM5 negated the impact of ZNF554 overexpression onβ-catenin and p-glycogen synthase kinase-3β(p-GSK-3β).Similarly,the deliberate activation of RBM5 reduced the increase inβ-catenin and p-GSK-3βcaused by the suppression of ZNF554.In vitro experiments showed that ZNF554 overexpression-induced decreases in cell proliferation and migration were counteracted by RBM5 knockdown.Additionally,when RBM5 was overexpressed,it hindered the improvements in cell proliferation and migration caused by reducing the ZNF554 levels.Conclusion:ZNF554 functions as a tumor suppressor in UCEC.Furthermore,ZNF554 regulates UCEC progression through the RBM5/WNT/β-catenin signaling pathway.ZNF554 shows a promise as both a prognostic biomarker and a therapeutic target for UCEC.