波形蛋白在不同病理亚型肾癌组织中的表达及其临床意义

目的:检测波形蛋白(vimentin)在不同病理类型肾细胞癌(RCC)组织中的表达并探讨其临床意义.方法:采用免疫组化PV-6000法检测152例手术切除的RCC组织和12例癌旁肾组织中波形蛋白(vimentin)的表达,分析vimentin表达与RCC临床病理特征的相关性.结果:vimentin在正常肾组织的阳性表达率为8.3%,在RCC组织的阳性表达率为76.3%,组间差异有统计学意义(P<0.05).vimentin在透明性肾细胞癌(ccRCC)、乳头状肾细胞癌(PRCC)和嫌色性肾细胞癌(CRCC)组织中阳性表达率分别为88.6%(101/114)、72.2%(13/18)和11.8%(2/17),其在ccRCC和PRCC的阳性表达均高于CRCC,组间差异均具有统计学意义(P<0.05).vimentin阳性表达与ccRCC的组织学分级和临床分期无明显相关性(P>0.05).结论:波形蛋白的表达与肾癌病理类型关系密切,提示vimentin可辅助RCC的病理分型鉴别诊断.波形蛋白在透明性肾细胞癌普遍高表达,提示其在ccRCC的诊断上有一定意义,但尚不足以评估ccRCC的恶性程度.     

Survivin和Vimentin在肾透明细胞癌中的表达及临床意义

目的 探讨凋亡蛋白抑制因子生存素(Survivin)和波形蛋白(Vimentin)在肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)中的表达及临床意义.方法 采用免疫组化SP法检测56例ccRCC、25例癌旁组织及13例正常肾组织中Survivin和Vimentin的表达及定位情况.结果 Survivin在肾透明细胞癌中高表达,而在癌旁及正常组织中未见表达;其表达与ccRCC临床分期及病理分级呈正相关,差异有统计学意义(P＜0.01).Vimentin在正常肾组织及癌旁组织中亦未见表达,而在肾透明细胞癌中高表达,且与临床分期及病理分级呈正相关,差异有统计学意义(P＜0.01).并且两个因子的表达呈显著正相关(r=0.566,P＜0.01).结论 Survivin和Vimentin在ccRCC组织中高表达,共同促进了ccRCC的发生、发展、侵袭与转移,联合两因子检测可为ccRCC的预后评估提供重要的信息.     

组氨酸三聚体核苷结合蛋白1在肾透明细胞癌中的表达及意义

目的:通过检测组氨酸三聚体核苷结合蛋白1(histidine triad nucleotide-binding protein1,HINT1)mRNA及其蛋白质在肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)中的表达情况,探讨其与临床病理特征的关系。方法:应用实时荧光定量(real-time fluorescence quantitative,RFQ)-PCR和免疫组织化学分别从mRNA和蛋白质水平检测HINT1在36例新鲜ccRCC组织、37例新鲜正常肾组织(其中29例配对)、30例石蜡包埋ccRCC组织与12例石蜡包埋正常肾组织中的表达水平,并分析其与临床病理特征间的关系。结果:HINT1mRNA在29例配对ccRCC和癌旁正常肾组织中的表达量分别为0.209±0.033和0.733±0.136(P<0.001),在36例ccRCC和37例癌旁正常肾组织中的表达量分别为0.245±0.035和0.694±0.108(P<0.001)。HINT1蛋白在癌旁正常肾组织中的阳性表达率为100%(12/12),弥漫性分布于细胞核及细胞质中,其中近曲小管表达强度高于远曲小管。肾小球基膜、肾小囊壁层上皮细胞可见HINT1蛋白部分表达,其强度低于肾小管,肾间质未见阳性表达。HINT1蛋白在ccRCC中的阳性表达率为60%(18/30),同样分布于细胞核及细胞质,癌组织间质阴性(P<0.01)。HINT1在mRNA和蛋白质水平表达趋势一致,在ccRCC中表达水平低于癌旁正常肾组织,并且在T3-4晚期肿瘤中表达量低于T1-2早期肿瘤中的表达量(P<0.05);但2者在不同Fuhrman核分级、性别、年龄和肿瘤大小中的表达差异无统计学意义(P>0.05)。结论:HINT1可能在ccRCC发病机制中发挥肿瘤抑制作用,其表达异常可能与翻译水平之前的异常调控有关。推测HINT1mRNA和蛋白质的异常表达可作为ccRCC发生和进展的预后指标。     

CD248在肾透明细胞癌中的表达及临床意义CSCD

目的探讨内皮唾酸蛋白(CD248)在肾透明细胞癌(ccRCC)组织中的表达及临床意义。方法收集115例ccRCC组织及对应癌旁组织的石蜡标本,同时收集其中17例ccRCC组织及对应癌旁组织的新鲜标本。qRT-PCR检测17例ccRCC组织及对应癌旁组织中CD248 mRNA的表达;免疫组织化学法检测115例ccRCC患者石蜡标本的CD248蛋白水平,分析CD248表达与ccRCC临床病理指标及预后的相关性。结果ccRCC组织中的CD248阳性表达免疫组织化学反应强度显著强于癌旁组织,差异有统计学意义(P=0.0061),且CD248蛋白阳性信号定位于细胞核及细胞质。CD248 mRNA在癌旁组织中的表达水平明显低于ccRCC组织(P=0.0026)。CD248高表达组组间总生存率明显低于CD248低表达组(44.4 vs.57.2月,P=0.017)。结论CD248的表达可能参与ccRCC的发生发展。与癌旁组织比较,CD248在ccRCC组织中高表达;CD248高表达的ccRCC患者术后生存时间较短。     

Vimentin在肾透明细胞癌中的表达及临床意义

目的:探讨波形蛋白(Vimentin)在人肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)和癌旁组织样本中的表达情况,并分析其与 ccRCC 临床分期、病理分级和侵袭转移等恶性生物学特性的关系及临床意义。方法:Western blot方法检测29例新鲜肾透明细胞癌及相应癌旁组织中Vimentin的表达;免疫组织化学方法检测120例肾透明细胞癌及相应癌旁组织中Vimentin的表达情况,分析Vimentin在肾透明细胞癌组织中及癌旁正常肾脏组织中的表达水平差异及Vimentin的表达与肿瘤大小、临床分期、组织病理分级、远处转移的关系。结果:Vimentin在肾透明细胞癌及相应癌旁组织中均有表达,前者的表达水平较后者显著升高（P＜0.01）;免疫组织化学显示肾透明细胞癌组织中,Vimentin的阳性表达率高达90.0%,与癌旁组织存在明显差异（P＜0.01）;Vimentin在肾透明细胞癌组织中以胞膜表达为主,其表达随肾癌的临床分期、肾包膜受侵、癌栓形成或远处转移有升高趋势,但组间比较经统计学分析均无显著性差异。结论:Vimentin在肾透明细胞癌组织中的表达明显高于癌旁正常肾组织,提示Vimentin在肾透明细胞癌的发生发展过程中发挥重要的作用。Vimentin在肾透明细胞癌组织中过表达,与肾癌的发展密切相关,可以作为肾细胞癌早期诊断、预后和复发评估的参考指标之一。     

肾透明细胞癌组织中NSUN2的表达及其与临床病理特征和预后的关系

目的:检测NSUN2在肾透明细胞癌(ccRCC)组织中的表达,分析其与临床病理特征、预后之间的关系。方法:收集ccRCC组织蜡块标本及对应的癌旁组织蜡块标本各95例,采用免疫组化法检测NSUN2在两组标本中的表达水平,分析NSUN2与ccRCC患者的临床病理特征关系。采用Kaplan-Meier法绘制生存曲线,使用Log-rank检验进行单因素生存分析,Cox回归模型进行多因素生存分析,分析NSUN2表达与ccRCC患者预后之间的相关性。结果:NSUN2在癌组织的高表达率为69.5%,在癌旁组织中的高表达率为15.8%,两组差异有统计学意义(P<0.05)。NSUN2高表达与肿瘤大小(P<0.05)、组织学分级(P<0.05)、淋巴转移(P<0.05)及临床分期有关(P<0.05)。Kaplan-Meier分析显示,NSUN2表达水平与ccRCC患者预后相关(P<0.05)。Cox回归模型显示,NSUN2表达是预测ccRCC预后的独立危险因素。结论:NSUN2在ccRCC组织中高表达,且与ccRCC的临床病理特征、预后相关,可作为ccRCC预后判断的...     

ALDH1A1在肾透明细胞癌组织中的表达及其临床意义

目的:探讨ALDH1A1(aldehyde dehydrogenase 1A1,ALDH1A1)在肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)组织中的表达及其与患者临床病理特征及预后的关系。方法:应用免疫组化SP法检测95例ccRCC和23例正常肾组织石蜡中ALDH1A1蛋白的表达水平。Realtime PCR和Western Blot检测20例新鲜ccRCC及癌旁组织中ALDH1A1 mRNA和蛋白的表达量。结果:ccRCC石蜡组织中ALDH1A1表达高于正常肾组织,两者之间差异有统计学意义(P<0.05)。新鲜ccRCC组织中ALDH1A1 mRNA及蛋白的表达水平较癌旁正常肾组织显著升高。ALDH1A1表达与肿瘤大小、临床分期、血管侵袭及复发相关(P<0.05)。Log-rank分析进一步表明ALDH1A1高表达提示不良预后。结论:ALDH1A1的表达与ccRCC的生物学行为关系密切,可能为ccRCC的治疗提供新的靶点。     

基于TCGA数据库分析CRB3在肾透明细胞癌（ccRCC）中的表达及其与预后的关系

目的:通过研究细胞极性蛋白CRB3（Crumbs 3）在肾透明细胞癌（clear cell renal cell carcinoma,ccRCC）组织中的表达,阐明其对ccRCC早期诊断及预后的作用。方法:运用癌症基因组图谱（TCGA）数据库和免疫组织化学（immunohistochemistry,IHC）技术分析CRB3在ccRCC组织和正常肾组织的表达情况,通过LinkedOmics和GEPIA阐述CRB3表达与ccRCC临床病理学参数的相关性及对预后的影响。结果:CRB3 mRNA在ccRCC中低表达,并且与ccRCC病理分级、TNM分期呈负相关,并与人种相关。TCGA结果显示CRB3低表达是影响ccRCC患者总生存率和无病生存率的独立预后因素（P<0.05）。IHC结果证实,与正常肾组织相比,ccRCC中CRB3的蛋白表达量显著降低（P<0.05）。结论:在ccRCC中,CRB3低表达是一种重要的不良预后指标,可以作为预测患者转移发生、判断预后的有效生物标志物。     

肾透明细胞癌预后相关蛋白标志物的研究进展

肾细胞癌(renal cell carcinoma,RCC)是人类最常见的泌尿系统恶性肿瘤之一,其最常见的病理类型为肾透明细胞癌（clear cell renal cell carcinoma,ccRCC）。近年来,国内外诸多研究报道多种蛋白的表达与ccRCC的预后密切相关。本文就这些蛋白标志物在判断ccRCC患者预后方面的重要作用做一综述。     

肾透明细胞癌组织40例En-1相关基因表达意义

目的 检测同源异型盒基因家族中En-1及其相关基因FoxO1和Sirt3在肾透明细胞癌(ccRCC)不同病理分级中的表达情况,为ccRCC的治疗提供新的思路.方法 收集2017-07-01-2018-10-31宁夏医科大学总医院术后病理诊断为ccRCC组织及癌旁组织(距癌组织>2 cm)40例,根据病理Fuhrman分...     

Circular RNA hsa_circ_001895 serves as a sponge of microRNA‐296‐5p to promote clear cell renal cell carcinoma progression by regulating SOX12

There is an urgent need to find novel potential therapeutic targets for the diagnosis and treatment of clear cell renal cell carcinoma (ccRCC) due to its highly invasive ability as a common urological malignant tumor. Circular RNAs (circRNAs) have been indicated as potentially critical mediators in various types of tumor progression. We first used qRT‐PCR analysis to find dysregulated circRNAs in ccRCC. A novel circRNA, hsa_circ_001895, was upregulated in ccRCC specimens and associated with metastatic properties of ccRCC. However, the tumorigenic mechanism of hsa_circ_001895 on ccRCC is yet to be found. We first indicated that hsa_circ_001895 predicted a poor prognosis in ccRCC patients. Additionally, overexpression of hsa_circ_001895 not only promoted cell proliferation, invasion and migration of ccRCC, but also inhibited cell apoptosis, whereas hsa_circ_001895 knockdown reversed the effect on ccRCC progression. In vivo s.c. xenotransplanted tumor model also showed that silencing hsa_circ_001895 could suppress in vivo ccRCC growth. Mechanistically, hsa_circ_001895 directly binds with microRNA (miR)‐296‐5p and inhibits its expression. Moreover, sex determining region Y (SRY)‐box 12 (SOX12) was identified as a target of miR‐296‐5p, the expression of which was suppressed by miR‐296‐5p. Notably, the inhibitory effect of hsa_circ_001895 on ccRCC progression was reversed by miR‐296‐5p inhibitor. In general, our findings indicated that hsa_circ_001895 may sponge miR‐296‐5p and promote SOX12 expression, which is the underlying mechanism of hsa_circ_001895‐induced ccRCC progression.     

CircPVT1 promotes progression in clear cell renal cell carcinoma by sponging miR-145-5p and regulating TBX15 expression

Emerging evidence revealed that circular RNAs (circRNAs) play significant roles in regulating tumorigenesis and cancer progression. However, few circRNAs were well characterized in clear cell renal cell carcinoma (ccRCC). We found that circPVT1 was significantly upregulated in ccRCC tissues and positively associated with the clinical stage. The Area Under Curve of tissue and serum circPVT1 expression in ccRCC were 0.93 and 0.86, respectively. Importantly, we demonstrated that circPVT1 promoted ccRCC growth and metastasis in vitro and in vivo. We also found that circPVT1 directly binds to miRNA-145-5p via the Biotin-labelled miRNA pulldown assay and dual-luciferase reporter assay, and miR-145-5p inhibitor significantly attenuated the effect of circPVT1 knockdown on ccRCC cells. Moreover, through RNA sequencing and bioinformatics analysis, we demonstrated that TBX15 was regulated by the circPVT1/miR-145-5p axis and predicted poor prognosis in ccRCC. These findings suggest that circPVT1 promotes ccRCC growth and metastasis through sponging miR-145-5p and regulating downstream target TBX15 expression. The circPVT1/miR-145-5p/TBX15 axis might be a potential diagnostic marker and therapeutic target in ccRCC.     

肾细胞癌的分子遗传学改变研究进展

肾细胞癌（RCC）是肾癌最重要的一种病理分型,其中又以透明细胞性肾细胞癌（ccRCC）最常见,其次是乳头状肾细胞癌（PRCC）和嫌色性肾细胞癌（ChRCC）。随着二代测序技术的普遍运用,关于ccRCC中VHL、PBRM1等常见突变基因以及3p缺失等拷贝数变异（CNV）的研究越来越多;另外,PRCC和ChRCC中的基因突变和CNV研究也不断出现。本文将对ccRCC、PRCC和ChRCC等几种常见RCC的基因突变、CNV和基因融合等分子遗传学改变研究进展作一综述。     

Silencing of lncRNA AFAP1-AS1 Inhibits Cell Growth and Metastasis in Clear Cell Renal Cell Carcinoma

The lncRNA AFAP1-AS1, oriented from an antisense direction to the protein-coding gene AFAP1 in the opposite strand, was upregulated in a variety of tumors and associated with poor prognosis, including lung cancer, breast cancer, ovarian cancer, and so on. However, the biological role of AFAP1-AS1 in clear cell renal cell carcinoma (ccRCC) is still unknown. We observed that AFAP1-AS1 expression was significantly upregulated in ccRCC tissues and that patients with high-level expression of AFAP1-AS1 had a shorter overall survival. Knockdown of AFAP1-AS1 markedly suppressed the progression of proliferation, invasion, migration, and EMT in ccRCC cells. Downregulation of AFAP1-AS1 resulted in an increase in E-cadherin and a decrease in vimentin. Noticeably, we found that PTEN has a negative correlation with the lncRNA AFAP1-AS1 expression. Further studies verified that PTEN deficiency effectively attenuated the ability of AFAP1-AS1 in promoting ccRCC cell proliferation, invasion, migration, and EMT. Moreover, the similar biological response of silencing AFAP1-AS1 was observed in our ccRCC mice model. Knockdown of AFAP1-AS1 evidently suppressed tumor growth. Taken together, our results provide the evidences that silencing of AFAP1-AS1 inhibits cell proliferation, EMT, and metastasis through PTEN-dependent signaling, and our findings elucidate a novel potential therapeutic target or biomarker for the treatment of ccRCC.     

SOX6 represses tumor growth of clear cell renal cell carcinoma by HMG domain-dependent regulation of Wnt/β-catenin signaling

Sex-determining region Y box (SOXs) are expressed in various cells and control cell fate and differentiation in a multitude of physiologic processes. SOX6, a main representative of SOXs, is involved in the regulation of carcinogenesis in various human malignancies. However, the role of SOX6 in clear cell renal cell carcinoma (ccRCC) remains unclear. In this study, SOX6 expression in ccRCC and its clinical significance were investigated. In vitro and in vivo assays were used to explore the tumor-related function and the underlying molecular mechanism of SOX6 in ccRCC. We confirmed that SOX6 was frequently downregulated in ccRCC tissues and cell lines. Besides, downregulation of SOX6 was significantly associated with larger tumor sizes, advanced tumor stage, higher Fuhrman grades, and its expression could act as an independent prognostic factor for ccRCC (hazards ratio = 0.590, P = .026). Gain/loss-of-function experiments demonstrated that SOX6 could remarkably inhibit tumor cell growth and foci formation in vitro and xenograft tumorigenesis in vivo, respectively. Mechanistically, SOX6 could influence cell cycle by regulating the G1/the S phase transition and had an inhibitory effect on Wnt/β-catenin signaling as well as its target genes, c-Myc and cyclin D1. Interesting, the tumor-suppressive function of SOX6 was proved to be dependent on its specific high-mobility-group (HMG) domain. In general, our findings indicated that SOX6 was a novel tumor suppressor and prognostic biomarker in ccRCC. SOX6 could inhibit tumor growth by negatively regulating the Wnt/β-catenin signaling pathway in an HMG domain-dependent manner in ccRCC, which might provide a novel therapeutic approach for ccRCC.     

Transmembrane protein ADAM29 facilitates cell proliferation,invasion and migration in clear cell renal cell carcinoma

Abnormal expression of ADAM29 has been frequently reported in several cancers, however, its role in clear cell renal cell carcinoma (ccRCC) has not evaluated in detail. Herein, we attempt to determine the biological role and the action mechanism of ADAM29 in ccRCC. Bioinformatics analysis based on the ccRCC RNA-Seq dataset from TCGA database revealed that ADAM29 was up-expressed in ccRCC tissues by comparison with normal tissues. And a significant increase of ADAM29 expression was also observed in 3 ccRCC cell lines (UT33A, Caki-1, and786-O) in comparison with normal cell line. Besides, high level of ADAM29 was found to be connected with the poor prognosis and could be considered as an independent prognosticator for patients with ccRCC. Furthermore, functional experiments in vitro demonstrated that ADAM29 promoted the growth, invasion and migration of ccRCC cells. Moreover, Western blot assays indicated that ADAM29 was positively correlated with the level of proliferation-related proteins Cyclin D1 and PCNA and motion-related proteins MMP9 and Snail. Our data indicate that ADAM29 acts as an oncogene that increases tumour cells proliferation, invasion and migration partly by regulating the expression of Cyclin D1/PCNA/MMP9/Snail, suggesting that ADAM29 may become a prognosticator and therapeutic candidate for ccRCC.     

Silent information regulator 2 promotes clear cell renal cell carcinoma progression through deacetylation and small ubiquitin-related modifier 1 modification of glucose 6-phosphate dehydrogenase

The regulatory relationship between silent information regulator 2 (SIRT2) and glucose 6-phosphate dehydrogenase (G6PD) in clear cell renal cell carcinoma (ccRCC) is still unclear. The present study aimed to explore the function of SIRT2 and its regulatory effect on G6PD in ccRCC. The Cancer Genome Atlas data mining of SIRT2 was first analyzed. Quantitative real-time PCR and western blot analyses were used to assess the mRNA and protein expression levels, respectively. Cell viability, colony formation, cell cycle, cell apoptosis, and TUNEL assays and EdU staining were used to investigate the roles of SIRT2 in ccRCC proliferation and apoptosis. The coimmunoprecipitation (Co-IP) assay was used to analyze the association between SIRT2 and G6PD in ccRCC cells. Quantitative Co-IP assay was used to detect the levels of G6PD ubiquitination and small ubiquitin-related modifier 1 (SUMO1). An in vivo experiment was also carried out to confirm in vitro findings. The results indicated that SIRT2 promoted ccRCC proliferation and inhibited apoptosis by regulating cell cycle and apoptosis related proteins. Silent information regulator 2 interacted with G6PD, facilitated its activity through deacetylation, and increased its stability by reducing its ubiquitination and enhancing its SUMO1 modification. Silent information regulator 2 also promoted ccRCC tumor development in vivo. Taken together, the present study indicated that SIRT2 promoted ccRCC progression by increasing G6PD activity and stability, and it could be a potential new diagnostic and therapeutic target for ccRCC.     

转录因子BATF3通过调控波形蛋白促进肾透明细胞癌细胞的恶性生物学行为

目的：探讨碱性亮氨酸拉链转录因子ATF样蛋白3（BATF3）在肾透明细胞癌（ccRCC）组织中的表达及其调控ccRCC细胞恶性生物学行为的分子机制。方法：收集2019 年3月至2022 年1月间在中国人民解放军联勤保障部队第九八〇医院手术切除的64例ccRCC组织和相应癌旁组织,qPCR 法检测ccRCC组织、癌旁组织和肾癌ACHN、786-O细胞中BATF3 mRNA的表达,免疫组化检测ccRCC 组织、癌旁组织中BATF3蛋白的表达,并分析其与临床病理特征的关系。构建BATF3敲减及过表达质粒,分别转染786-O、ACHN 细胞,通过MTS 法、Transwell 实验检测BATF3 对786-O 或ACHN 细胞增殖、迁移、侵袭能力的影响,qPCR 法检测敲减或过表达BATF3 对786-O 或ACHN 细胞EMT 相关基因表达的影响,CHIP、双荧光素酶报告基因实验检测BATF3是否与波形蛋白（VIM）启动子区结合并调控其转录,MTS法、Transwell 实验检测同时过表达BATF3及敲减VIM对786-O细胞增殖、迁移、侵袭能力的影响。结果：与癌旁组织比较,ccRCC 组织中BATF3的mRNA和蛋白均呈高表达(均P<0.01),并且BATF3 mRNA 与ccRCC 的分化程度和TNM分期密切关联（均P<0.01）;与正常肾上皮细胞293T相比,BATF3 在ACHN 及786-O细胞中均呈高表达（均P<0.01）。敲减BATF3 表达均能明显抑制786-O 细胞的增殖、迁移、侵袭能力(均P<0.01),过表达BATF3则均能促进ACHN细胞的增殖、迁移和侵袭能力（均P<0.01）,敲减或过表达BATF3能抑制786-O 细胞或促进ACHN细胞的EMT相关基因的表达（均P<0.01）。BATF3可与VIM启动子区的位点结合,直接调控VIM的转录表达。同时过表达BATF3及敲减VIM可逆转过表达BATF3对786-O 细胞增殖、迁移、侵袭能力的影响。结论：BATF3在ccRCC 组织中呈高表达,并与其分化程度和TNM 分期密切关联;BATF3 通过调控VIM 的表达影响ACHN、786-O 细胞的恶性生物学行为,其可作为临床治疗ccRCC的潜在靶点。