乳腺癌患者病理特征与Bcl-2、CXCL13、PAX8表达情况的关系分析

目的分析乳腺癌患者病理特征与B细胞淋巴瘤/白血病-2基因(Bcl-2)、趋化因子配体13(CXCL13)、配对盒基因8抗体(PAX8)表达情况的关系。方法收集2021年1月至2023年1月该院收治的160例乳腺癌患者临床资料。采用免疫组化法对其癌组织与癌旁组织Bcl-2、CXCL13、PAX8表达情况进行检测,并分析3项指标与患者病理特征的关系。结果与癌旁组织比较,癌组织Bcl-2、CXCL13、PAX8阳性率更高,差异有统计学意义(P<0.05)。与雌激素受体(ER)阴性、肿瘤最大径≥3 cm、孕激素受体(PR)阴性患者比较,ER阳性、肿瘤最大径<3 cm、PR阳性患者中Bcl-2高表达占比更高,差异有统计学意义(P<0.05);与无淋巴结转移、Ⅰ~Ⅱ期患者比较,淋巴结转移、Ⅲ~Ⅳ期患者中CXCL13高表达占比更高,差异有统计学意义(P<0.05);与Ⅰ~Ⅱ期、高/中分化、无淋巴结转移患者比较,Ⅲ~Ⅳ期、低分化、有淋巴结转移患者中PAX8高表达占比更高,差异有统计学意义(P<0.05)。ER、PR表达情况与Bcl-2表达情况呈正相关(P<0.05),肿瘤最大径与Bcl-2表达情况呈负相关(P<0.05);临床分期、淋巴结转移情况与CXCL13、PAX8表达情况呈正相关(P<0.05);分化程度与PAX8表达情况呈负相关(P<0.05)。结论乳腺癌患者Bcl-2、CXCL13、PAX8表达情况对疾病的发生和发展具有明显影响,有望成为评估乳腺癌患者病情严重程度的标志物。

血清胸腺活化调节趋化因子、CXC亚家族趋化因子13与弥漫大B细胞淋巴瘤患者化疗疗效和预后的关系研究

目的探讨血清胸腺活化调节趋化因子(TARC)、CXC亚家族趋化因子13(CXCL13)与弥漫大B细胞淋巴瘤(DLBCL)患者化疗疗效和预后的关系。方法选取2017年1月至2020年6月该院收治的285例DLBCL患者(DLBCL组)及同期健康体检者160例(对照组)作为研究对象,DLBCL患者接受利妥昔单抗+环磷酰胺+多柔比星+长春新碱+泼尼松(R-CHOP)相关化疗方案,根据化疗效果分为有效组和无效组。比较DLBCL组与对照组、有效组与无效组血清TARC、CXCL13水平;随访统计DLBCL患者3年总生存期(OS)及无进展生存期(PFS),采用Logistic回归分析DLBCL患者预后及疾病进展的影响因素,并构建回归预测模型,采用受试者工作特征(ROC)曲线分析其预测评估效能。采用Kaplan-Meier生存曲线分析不同水平血清TARC、CXCL13与DLBCL患者3年OS及PFS的关系。结果DLBCL组治疗前血清TARC、CXCL13水平高于对照组(P<0.05)。DLBCL患者化疗有效率为85.26%,无效组治疗前血清TARC、CXCL13水平高于有效组(P<0.05)。多因素Logistic分析结果显示,Ann Arbor分期Ⅲ/Ⅳ期、TARC高表达及CXCL13高表达均是DLBCL患者预后的独立危险因素(P<0.05)。乳酸脱氢酶水平升高、TARC高表达及CXCL13高表达均是DLBCL患者疾病进展的独立危险因素(P<0.05)。以上述影响因素构建回归预测模型,ROC曲线分析显示,预测模型预测预后的曲线下面积(AUC)为0.874,预测疾病进展的AUC为0.911。TARC低表达患者的3年OS、PFS优于高表达患者(P<0.05),CXCL13低表达患者的3年OS、PFS优于高表达患者(P<0.05)。结论DLBCL患者血清TARC、CXCL13水平异常升高,血清TARC、CXCL13低表达的DLBCL患者具有更好的化疗效果及预后。包括血清TARC、CXCL13在内的多因子预测模型对DLBCL患者预后具有较高的评估效能。

ILC2和MDSC及其相关细胞因子IL-13和iNOS在宫颈癌中的表达及其列线图模型的构建和评价

目的:研究2型固有淋巴样细胞(ILC2)和髓源性抑制细胞(MDSC)及其相关细胞因子IL-13和诱导型一氧化氮合酶(iNOS)在宫颈癌(CC)中的表达,并基于其构建CC发病风险的列线图预测模型。方法:采集2022年5月至2024年1月在新疆医科大学第一附属医院手术切除的40例CC组织及100例外周血作为CC组,选取同期收治的30例子宫肌瘤经CC筛查为阴性的宫颈组织和100例正常健康个体外周血作为对照组。用多重免疫荧光技术(mIF)及免疫组化染色(IHC)法检测两组组织中ILC2和MDSC细胞浸润及其相关细胞因IL-13和iNOS的表达;使用流式细胞术和ELISA技术分别检测两组外周血中ILC2和MDSC及IL-13和iNOS的表达差异;通过Person相关性分析评估其相关性;使用单因素和多因素Logistic分析来确定ILC2和MDSC及IL-13和iNOS是否为CC发病的独立危险因素,再利用R软件建立免疫预测模型,使用ROC曲线下面积(AUC值)、Hosmer-Lemeshow检验、校准曲线、临床决策曲线和临床影响曲线来分别评估模型。结果:CC组中ILC2及MDSC及其相关细胞因子IL-13和iNOS均高于对照组(均P<0.05),且其均呈正相关(均P<0.05);经过单因素及多因素Logistic回归分析显示,ILC2、MDSC及IL-13、iNOS均是CC发病的独立危险因素(均P<0.05);基于这些危险因素的CC发病列线图,经过验证提示,该列线图模型具有一定的临床实用价值。结论:ILC2和MDSC及其相关的细胞因子IL-13和iNOS在CC组织及外周血中均呈高表达,基于这些危险因素构建的预测模型具有良好的预测能力和一定的实用性,为CC的早期诊断和治疗提供了一种简便有效的辅助工具。

补肾痹通方联合骨髓间充质干细胞对损伤软骨细胞的保护机制及SOX9、MMP-13表达的影响

目的:探讨补肾痹通方(BSBT)联合骨髓间充质干细胞(BMSCs)对白介素(IL)-1β诱导的损伤软骨细胞的保护机制及性别决定区Y框蛋白9(SOX9)、基质金属蛋白酶13(MMP-13)表达的影响。方法:使用10 ng/ml的IL-1β建立损伤软骨细胞模型,实验分为对照组、模型组(IL-1β)、中药组(IL-1β+BSBT)、干细胞组(IL-1β+BMSCs)和联合组(IL-1β+BSBT+BMSCs);CCK-8法检测各组软骨细胞增殖情况;RT-qPCR法和Western blot法分别检测软骨细胞内SOX9、MMP-13、Ⅱ型胶原α1重组蛋白(COL2A1)、IL-10的基因和蛋白表达;ELISA检测各组细胞培养上清中肿瘤坏死因子-α(TNF-α)、IL-6、胰岛素样生长因子1(IGF-1)、碱性成纤维细胞生长因子(bFGF)的水平。结果:与模型组比较,各组软骨细胞活性显著增强(P<0.01),软骨细胞中的SOX9、COL2A1、IL-10的基因和蛋白水平显著升高(P<0.01),MMP-13的基因和蛋白水平明显降低(P<0.01),软骨细胞培养上清中TNF-α、IL-6的含量显著降低(P<0.01),IGF-1、bFGF的水平升高(P<0.01),其中联合组变化最明显(P<0.05)。结论:BSBT联合BMSCs可有效保护损伤软骨细胞,其机制可能是通过上调SOX9,下调MMP-13,抑制炎症,改善软骨细胞微环境,促进关节软骨的修复与再生。

人类恶性肿瘤靶向治疗的新希望—CDK12/CDK13

细胞周期的更替有赖于细胞周期蛋白依赖性激酶(CDKs),CDKs是重要的蛋白激酶家族,在调节细胞周期和调控基因转录方面具有关键的作用。其中CDK12/CDK13在DNA损伤应答、RNA剪接调控、转录及细胞周期调控等方面至关重要。近年发现CDK12/CDK13在多种癌症中表达异常或发生突变,在癌症发展中扮演着重要角色,已成为近年来研究的热点之一。本篇基于Google Scholar、PubMed和万方数据库,归纳总结了CDK12/CDK13的基本结构、生物学功能、与恶性肿瘤的相关性、以及针对性抑制剂的研究进展进行综述。为后续的靶向治疗新型靶点的研发和机制探讨提供方向,为恶性肿瘤的防治、诊断以及治疗提供新思路。

细胞周期蛋白O调控细胞周期蛋白依赖性激酶13对宫颈癌进展的影响

目的:探讨细胞周期蛋白O(CCNO)通过调控细胞周期蛋白依赖性激酶13(CDK13)进而影响宫颈癌细胞周期进展的分子机制。方法:通过生物信息学方法分析CCNO在不同癌症中的表达情况。利用免疫组化(IHC)检测CCNO在宫颈癌患者组织样本中的表达情况,利用蛋白质免疫印迹(WB)检测CCNO在宫颈癌细胞系中的表达情况。通过WB检测CCNO载体沉默效率,通过细胞克隆、胸腺嘧啶核苷类似物实验检测(EDU)和细胞计数试剂盒-8(CCK8)检测肿瘤样本(NC)和肿瘤沉默CCNO样本(si-CCNO)的细胞增殖情况,通过细胞迁移实验和流式细胞分析实验检测2组细胞迁移情况和细胞周期的变化,最后通过免疫共沉淀(CO-IP)实验检测CCNO是否调控细胞周期蛋白依赖性激酶13(CDK13)的表达。结果:CCNO在宫颈癌患者的组织样本中高表达;与对照组比较,si-CCNO组中CCNO表达量降低(P<0.05);与NC组比较,si-CCNO组的细胞增殖降低(P<0.05),si-CCNO组的细胞迁移数目减少(P<0.05);与NC组比较,si-CCNO组停滞在G1期细胞明显增多(P<0.05);与NC组比较,si-CCNO组的CDK13表达显著降低。结论:沉默CCNO能够抑制宫颈癌细胞的增殖、细胞周期进展和转移能力,并且CCNO能够调控CDK13进而对宫颈癌进展产生影响。

兔出血症病毒复制子的构建及其在RK-13细胞中的复制

为研究兔出血症病毒(RHDV)的复制机制、病毒与宿主之间的相互作用以及致病机制等,创建一个安全、有效的技术平台,在前期构建的RHDV侵染性克隆基础上,将病毒的衣壳蛋白编码区删除,保留了RHDV复制必需的所有蛋白酶基因和两端的非编码区,构建了RHDV复制子。试验结果证明,将该复制子RNA导入RK13细胞中后,能够进行高水平的复制和表达。

Interleukin-13 inhibits cytokines synthesis by blocking nuclear factor-κB and c-Jun N-terminal kinase in human mesangial cells

Objective： Monocytes/macrophages, proinflammatory cytokines and chemokines are important in the pathogenesis of glomerulonephritis. Interleukin （IL） -13 has been shown to exert potent anti-inflammatory properties. This study was designed to investigate the effect of IL-13 on the expression of proinflammatory cytokines, chemokines and profibrogenic cytokines and the involved molecular mechanism in cultured human mesangial cells （HMCs）. Methods： The expressions of proinflammatory cytokines, chemokines and profibrogenic cytokines were determined by ribonuclease protection assay （RPA）. Activity of nuclear factor-kappa B （NF-κB） and activa- tor protein-1 （AP-1） was examined by electrophoretic mobility shift assay （EMSA）. NF-κB subunit p65 nuclear transportation and c-Jun N-terminal kinase （JNK） activity were assayed by immunoblot. Results： Recombinant IL-13 inhibited tumor necrosis factor-α （TNF-u）, IL-1α, IL-1β, monocyte chemoattractant protein-1 （MCP-1）, IL-8, and transforming growth factor-β1 （TGF-β1） mRNA expressions in a dose-dependent manner. Lipopoly- sacchorides （LPS） dramatically increased NF-κB DNA binding activity of HMCs, which was inhibited by IL-13 in a dose-dependent manner. LPS-activated NF-κB contained p50 and p65 dimers, but not c-Rel subunit. IL-13 blocked LPS-induced NF-κB subunit p65. LPS stimulated JNK/AP-1 activation, which was inhibited by IL-13 in a dose-dependent manner. Conclusion： IL-13 inhibits proinflammatory cytokines, chemokines, and profibrogenic cytokines synthesis by blocking NF-κB and JNK/AP-1 activation. These observations point to the importance of IL-13 in the modulation of inflammatory processes in the renal glomerulus.

Construction of eukaryotic expression vector of human S100A13 gene and its effect on proliferation of human thyroid cancer cell line TT

Objective To investigate the effect of exogenous S100A13 gene overexpression on the proliferation of human thyroid cancer cell line TT.Methods The recombinant ORF of S100A13 tagged with six histidines at the 5' end was subcloned into the pcDNA3.2/V5/GW/D-TOPO vector and sequenced.The eukaryotic expression plasmid pcDNA3.2/V5 /GW/D-S100A13 and empty vector pcDNA3.2/V5/GW/D were transfected into TT cells.The positive clones were selected by G418.The expressions of S100A13 mRNA and protein were detected by real time reverse transcription-polymerase chain reaction(RT-PCR) and Western blot.The effect of S100A13 on cell proliferation and cell cycle was evaluated by cell growth curve,MTT colorimetric assay and flow cytometry.Results S100A13 gene tagged with six histidines at the 5 ' end was confirmed to be inserted into the pcDNA3.2/V5/GW/D vector correctly.TT-S100A13-V5 cells,which over-expressed S100A13,were constructed successfully.TT-S100A13-V5 cells grew much faster than TT-V5 and TT cells(P <0.001).The proportions of both S and G2/M phase cells were significantly higher in TT-S100A13-V5 cells than those in TT-V5 and TT cells(P <0.001).Conclusion The eukaryotic expression vector containing human S100A13 gene has been successfully constructed,which highly expresses S100A13 in TT cells.Exogenous S100A13 gene overexpression accelerates TT cell proliferation and drives the cell cycle progression of TT cells from G0/G1 phase to S and G2/M phases.

IL-13Ra2-and glioma stem cell-pulsed dendritic cells induce glioma cell death in vitro

Objective Gliomas are the most common malignant tumors in the central nervous system.Despite multiple therapies including surgery,chemotherapy,and radiotherapy,the prognosis of patients remains poor.Immunotherapy is an alternative method of treating glioma,and the use of dendritic cell vaccines is one of the promising treatment options.However,there is no specific tumor cell antigen that can trigger dendritic cells(DCs).IL-13Ra2 is a specific antigen expressed in glioma cells;in the current study,we have attempted to explore whether IL-13Ra2 could be the antigen that triggers DCs and to envisage its application as potential therapy for glioma.Methods The expression of IL-13Ra2 was detected in U251 glioma cell lines and primary glioma tissues using different methods.DCs from human blood were isolated and pulsed with recombinant IL-13Ra2,following which the cytotoxicity of these DCs on glioma cells was detected and analyzed.Results About 55.9% human glioma tissue cells expressed IL-13Ra2,while normal brain tissue cells did not show any expression.DC vaccines loaded with IL-13Ra2,glioma cell antigen,and brain tumor stem cell(BTSC) antigen could significantly stimulate the proliferation of T lymphocytes and induce cell death in the glioma tissue.Compared to other groups,DC vaccines loaded with BTSC antigen showed the strongest ability to activate cytotoxic T lymphocytes(CTLs),while the glioma cell antigen group showed no significant difference.Conclusion IL-13Ra2,which is expressed in gliomas and by glioma stem cells,as well as IL-13Ra2 could prove to be potential antigens for DC vaccine-based immunotherapy.

Tinospora crispa extract inhibits MMP-13 and migration of head and neck squamous cell carcinoma cell lines

Objective: To investigate the effect of Tinospora crispa(T. crispa) extract on matrix metalloproteinase 13(MMP-13) expression and cell migration. Methods: The cytotoxicity of T. crispa extract was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on head and neck squamous cell carcinoma(HNSCC) cell lines. The effect on expression of MMP-13 was analysed by RT-PCR and ELISA. The migration was assessed by wound healing assay. Results: MMP-13 m RNA was highly expressed in the metastatic human HNSCC cell lines, HN22 and HSC-3. T. crispa extract at a concentration of 100.0 μg/m L caused about 50% reduction of cell survival. T. crispa extract at a non-toxic concentration of 12.5, 25.0 and 50.0 μg/m L signii cantly suppressed MMP-13 m RNA expression and secreted MMP-13 in both HN22 and HSC-3. The expression of tissue inhibitors of metalloprotease by HSC-3 cells was attenuated by 25.0 and 50.0 μg/m L of T. crispa extract. Addition of the extract to cells in a wound healing assay showed inhibition of cell migration by HN22 cells. Conclusions: These data suggest that T. crispa could be considered as a potential therapeutic drug to prevent metastasis of HNSCC.

Glycosylation-independent binding to extracellular domains 11-13 of mannose-6-phosphate/insulin-like growth factor-2 receptor mediates the effects of soluble CREG on the phenotypic proliferation of vascular smooth muscle cells

Background The present study aimed to investigate the detailed mode and specific sites for their binding as well as the functional relevance of this binding in the phenotypic proliferation of vascular smooth muscle cells(SMCs). Methods CREG knocked-down SMCs were employed to evaluate the biological activity of wtCREG and mCREG.Expressions of SMC differentiation markers SM myosin heavy chain(SM-MHC),SM-actin,heavy caldesmon and myocardin were determined by Western blotting using specific antibodies. Cellular growth of SMCs was assessed by bromide dewuridine (BrdU) incorporation and cell cycle analysis on fluorescence-activated cell sorting(FACS).A solid-phase binding assay was used to study the binding of CREG to extracellular domains of M6P/IGF2R.The cellular co-localization of the two recombinant CREGs with M6P/IGF2R was detected on SMC surface by immunoprecipitation and immunofluorescence analysis.Results The molecular weight of wtCREG was around 30 kD while that of the mCREG was～25 kD.Treatment of wtCREG with PNGase F reduced its molecular weight from～30 kD to～25 kD,whereas PNGase F treatment had no effect on the molecular weight of mCREG.Both wtCREG and mCREG proteins enhanced SMC differentiation,inhibited BrdU incorporation,and arrested cell cycle progression when added to the culture medium.In CREG knocked-down SMCs,the amount of CREG detected by immunoblotting in M6P/IGF2R immunoprecipitates was significantly reduced when compared to normal cells.Both recombinant CREGs co-immunoprecipitated with M6P/IGF2R, although slightly reduced amount of the mutant CREG was detected in M6P/IGF2R immunoprecipitates.Immunostaining revealed that His-tagged CREGs co-localized with IGF2R on the cell surface in a glycosylation-independent manner.In vitro binding assay showed that CREGs bound to M6P/ IGF2R extracellular domains 7-10 and 11-13 in a glycosylation -dependent and -independent manner,respectively.Further blocking experiments using soluble M6P/IGF2R fragments and M6P/IGF2R neutralizing antibody indicated that the biological activities of recombinant CREGs in SMC growth and the up-regulation of SMC differentiation markers were all abolished by treatment with the M6P/IGF2R neutralizing antibody. However,although the growth inhibitory effect of wtCREG was nearly abolished by D7-10 or D11-13,the effect of mCREG was only reversed by Dll-13,indicating that the binding to domains 11-13 is required for CREG to modulate the proliferation of SMCs.Conclusions These data suggest that solubleCREG proteins can exert their biological function via binding to the extracellular domains 7-10 and 11-13 of cell surface M6P/IGF2R in both a glycosylation-dependent and -independent manner.

Regulation of mitochondrial carrier SLC25A13 on breast cancer cell cycle in vitro

Objective·To investigate the role of mitochondrial solute carrier family 25 member 13(SLC25A13)on breast cancer development.Methods·SLC25A13 mRNA and protein expressions in invasive breast cancer tissues and normal breast tissues were from The Cancer Genome Atlas(TCGA)breast cancer dataset.Survival analysis was conducted online by Kaplan-Meier software.MCF-7 cell line was used for in vitro cell assay.Knockdown of SLC25A13 and sirtuin 2(SIRT2)were conducted by siRNA transfection.Cell viability was measured with trypan blue exclusion.Cell cycle arrest was determined by flow cytometry.The mRNA expression of SLC25A13 and P27 were detected by quantitative PCR.The protein level of SLC25A13,P27 and SIRT2 were detected by Western blotting.Protein half-life of P27 was assessed by Western blotting after cycloheximide treatment.Results·SLC25A13 was up-regulated in invasive breast cancer tissues.High expression of SLC25A13 correlated with poor overall survival and breast cancer recurrence.SLC25A13 knockdown inhibited MCF-7 cell cycle progression.P27 and SIRT2 both accumulated after SLC25A13 knockdown.P27 accumulation resulted from prolonged protein half-life.Knockdown of SIRT2 restored cell cycle arrest as well as P27 accumulation caused by SLC25A13 silencing.Conclusion·High expression of SLC25A13 may promote cell cycle progression via SIRT2 in breast cancer development.

Two potentially specific but relevant patterns of proteomic change Response of SH-SY5Y cells to differentiation with retinoic acid followed by phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate, and susceptibility of differentiated cells to dopamine

Dopamine （DA） exposure at a dose of 100 pmol/L for 24 hours causes oxidative stress in SH-SY5Y cells with induction of neuronal differentiation by retinoic acid （RA,10 pmol/L,72 hours） followed by phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate （TPA,80 nmol/L,72 hours）. However,it remains unclear whether the alteration of phenotype observed in response to oxidative stress is associated with protein regulation in this cellular model for Parkinson＇s disease. The present study detected protein regulation affected by oxidative stress at a proteomic level：selection of differentially altered proteins using two dimensional difference in-gel electrophoresis and identification of these proteins using matrix assisted laser desorption/ionization time-of-flight mass spectrometry. The results demonstrated significant alterations in expression of six proteins in SH-SY5Y cells following the differentiation and fourteen proteins in the differentiated cells following the exposure,exemplified by an increase of tubulin alpha1 in the former but a decrease of tubulin alpha-ubiquitous chain in the latter. These results suggest that two potentially specific but relevant patterns of proteomic change may be produced in SH-SY5Y cells with the induction of differentiation by RA followed by TPA,and in the differentiated cells after DA exposure.

白细胞介素13调控人鼻黏膜上皮基底细胞增殖

目的 探究慢性鼻窦炎伴鼻息肉(CRSwNP)鼻黏膜上皮基底细胞增殖情况,以及白细胞介素13(IL-13)对基底细胞增殖的影响。方法 收集健康对照及CRSwNP患者鼻黏膜组织,并对组织进行切片、免疫荧光染色及real-time PCR检测。分离培养原代人鼻黏膜上皮细胞,经IL-13处理后利用real-time PCR结合免疫荧光法检测基底细胞增殖及相关基因的表达。结果 与对照组相比,CRSwNP患者上皮中表达肿瘤蛋白p63(TP63)的基底细胞数量显著增加;TP63、细胞增殖标志物Ki67(MKI67)、炎症因子IL-13在转录表达水平显著升高;IL-13的表达与TP63、MKI67的表达呈显著正相关。原代鼻黏膜上皮细胞经IL-13刺激,显著促进TP63+基底细胞的增殖,以及TP63、MKI67基因的表达。结论 CRSwNP鼻黏膜上皮基底细胞增殖显著增加可能与炎症因子IL-13的调控有关。

Replication of M13 single-stranded DNA bearing a sitespecific ethenocytosine lesion by Escherichia coil cell extracts

Previous investigation on the mutagenic effects of 3,N4-Ethenocytosine (εC), a nonpairing DNA lesion,revealed the existence of a novel SOS-independent inducible mutagenic mechanism in E. coli termed UVM for UV modulation of mutagenesis. To investigate whether UVM is mediated by an alteration of DNA replication, we have set up an in vitro replication system ill which phage M13 viral single-stranded DNA bearing a single site-specific (εC) residue is replicated by soluble protein extracts from E. coli cells. Replication products were analyzed by agarose gel electrophoresis and the frequency of translesion synthesis was determined by restriction endonuclease analyses. Our data indicate that DNA replication is strongly inhibited by εC, but that translesion DNA synthesis does occur in about 14% of the replicated DNA molecules. These results are very similar to those observed previously in vivo, and suggest that this experimental system may be suitable for evaluating alterations in DNA replication in UVM-induced cells.

TRIP13 is identified as a prognosis biomarker for renal clear cell carcinoma and promotes renal cell carcinoma cell proliferation, migration and invasion

This work aimed to discover new therapeutic targets in renal clear cell carcinoma by bioinformatics and detect the effect of candidate gene TRIP13 in renal cell carcinoma(RCC)cell proliferation,migration,and invasion.Differentially expressed mRNAs were screened based on The Cancer Genome Atlas(TCGA)-Kidney Renal Clear Cell Carcinoma(KIRC)databases,and functional enrichments,survival analysis,receiver operating characteristic curve(ROC),and Protein–Protein Interaction(PPI)protein interaction analysis were performed by R software to screen the candidate gene TRIP13.Then,the expression of candidate gene TRIP13 in 92 pairs of cancer and adjacent normal tissues of renal clear cell carcinoma patients were detected by qRT-PCR,western blotting,and immunochemical analysis.The TRIP13 level and clinicopathological characteristics of patients with renal clear cell carcinoma were analyzed.Using 186-O and ACHN RCC cell lines with TRIP13 overexpressing or downregulating,the effect of TRIP13 on cell viability and proliferation were detected by CCK8 and EdU staining,respectively.The migration and invasion were detected by Transwell assays.A total of 19858 differentially expressed genes,5823 differentially expressed genes,3657 up-regulated genes,and 2166 down-regulated genes were identified.TRIP13 was closed associated with cell cycle regulation,and survival and prognosis of renal clear cell carcinoma were selected as a candidate gene.The mRNA and protein levels of TRIP13 in cancer tissues were higher than that in adjacent normal tissues.TRIP13 level was significantly associated with tumor size,tumor stage,Fuhrman grade,and lymph node metastasis.TRIP13 overexpression significantly increased cell viability,proliferation,migration,and invasion,while downregulating of TRIP13 had opposite effects in both 186-O and ACHN cells.Therefore,TRIP13 promotes RCC proliferation and metastasis,which should be a novel biomarker for early diagnosis,treatment,and prognosis of RCC.

Chemokine Ligand 13 Expression is Abundant in the Tumor Microenvironment and Indicates Poor Prognosis of Kidney Clear Cell Carcinoma

The chemokine ligand 13-chemokine receptor 5(CXCL13-CXCR5)axis has been characterized as a critical tumor-promoting signaling pathway in the tumor microenvironment(TME)in multiple types of solid tumors.In this study,we analyzed the expression profile of CXCL13 in kidney clear cell carcinoma(KIRC)and its correlation with tumor-infiltrating immune cells(TIICs).A monoclonal antibody against CXCL13 with high affinity and purity was generated in our lab for western blot and immunohistochemistry(IHC).Bioinformatic analysis was performed based on bulk-seq data from the Cancer Genome Atlas(TCGA)-KIRC and single-cell RNA-seq data from scRNASeqDB and PanglaoDB.Results showed that high CXCL13 expression in TME was associated with shorter progression-free survival(PFS),disease-specific survival(DSS),and overall survival(OS).KIRC cell lines,as well as several other cancer cell lines,had negative CXCL13 expression.IHC staining from the Human Protein Atlas(HPA)and our tissue array indicated that CXCL13 might be mainly expressed by TIICs,but not KIRC tumor cells.CXCL13 expression was strongly and positively correlated withγδT cell abundance in TME.Besides,γδT cell infiltration was associated with poor survival of KIRC.Methylation 450k array data showed that CXCL13 promoter hypomethylation was common in TIICs.The methylation level of cg16361705 within the CXCL13 promoter might play an important role in modulating CXCL13 transcription.In conclusion,our study revealed that CXCL13 expression andγδT cell infiltration in TME is associated with unfavorable survival of KIRC.TIICs,most possiblyγδT cells,are the dominant source of CXCL13 in KIRC TME.

Irradiation Injury Temporarily Induces Enhancement of APN/CD13 Peptidase Activity on Aorta-gonads-mesonephros (AGM)-derived Stromal Cells

This study was designed to investigate the expression of aminopeptidase N （APN）/CD13 on intraembryonic AGM stromal cells, and the change of its enzymatic activity after irradiation injury. The expression of APN/CD13 on AGM stromal cells was assayed by RT-PCR and immunihistochemistry. After the stromal cells in AGM region were irradiated with 8.0 Gy of ^60Co T-rays, APN/CD13 enzymatic activity was measured by spectrophotometer at different time points. The result showed that AGM stromal cells strongly expressed APN/CD13. The enzymatic activity of APN/CD13 decreased temporarily after irradiation injury, then increased to higher level 4 h after irradiation, and it returned to the pre-irradiation level 24 to 48 h after the irradiation. The enzymatic activity of APN/CD13 was temporarily enhanced after irradiation injury, which might be one of the compensatory mechanisms that promote the hematopoietic recovery after irradiation.

Evaluation of Changes in Actin Filaments of RK13 Cells Infected with <i>Malassezia pachydermatis</i>

Background: Malassezia pachydermatis is the main causative agent of canine otitis and also of a myriad of dermatological problems in companion animals;its interaction mechanisms with host cells are still unclear. Objectives: To establish an in vitro infection model of M. pachydermatis-exposed RK13 cells in order to evaluate cell morphological changes as well as changes in the structure of actin filaments. Methods: Cultures of RK13 cells were infected with M. pachydermatis, alterations caused by the yeast were evaluated by optical and fluorescence microscopy. Results: M. pachydermatis adheres itself to the cell and produces the formation of multiple agglomerates that cause changes in cell morphology, formation of cell aggregates in overlays, presence of syncytia and destruction of cell culture structure. The damaged cells presented changes in the actin filaments consisting of thickening of the cell cortex and loss of stress fibers. On the other hand, the formation of perinuclear actin rings in the yeasts was observed. Conclusions: An in vitro infection model was established with M. pachydermatis and alterations in cell morphology were observed consisting of changes in the structure of the actin filaments, overgrowth of the cells and the presence of syncytia.