沉默EMMPRIN对肾透明细胞癌增殖的影响

目的:探讨细胞外基质金属蛋白酶诱导因子（EMMPRIN）在肾透明细胞癌增殖中的作用。方法:利用免疫组化、实时定量PCR(RT-PCR)和蛋白印迹(Western blot)等方法检测肾癌组织与肾透明细胞癌细胞株中EMMPRIN的表达情况。通过小分子干扰核糖核酸（siRNA）EMMPRIN在786-O细胞中的表达,研究EMMPRIN下调后对786-O肾癌细胞株生物学功能的影响。结果:EMMPRIN在肾癌组织中高表达,肾癌细胞系中EMMPRIN的表达高于正常的肾小管上皮细胞。通过小干扰RNA来敲除EMMPRIN的表达,并影响肾透明细胞癌的增殖。结论:EMMPRIN在肾肿瘤组织和肾癌细胞株中皆呈现出过表达的状态。下调其表达可以降低肾细胞癌的增殖水平,表明EMMPRIN的高表达可能是肾细胞癌患者预后较差的指标,可被视为肾癌治疗的潜在靶点。     

桧木醇诱导人肾透明细胞癌786-O细胞凋亡及其机制研究*

目的:探讨桧木醇（hinokitiol ）对人肾透明细胞癌786-O细胞株的增殖抑制作用及诱导凋亡效应,初步阐明其机制。方法:采用CCK-8 法检测桧木醇对肾癌786-O细胞增殖的抑制作用;流式细胞术检测细胞凋亡情况;转染EGFP-LC3 后镜下观察细胞自噬状态;采用Western blot对细胞Caspase- 3 剪切体、LC3 和P62蛋白表达量进行检测。结果:桧木醇可以显著抑制肾癌786-O细胞增殖,通过激活Caspase途径诱导细胞凋亡。桧木醇诱导肾癌786-O细胞发生自噬,LC3 蛋白表达显著上调,而P62蛋白表达下调。结论:桧木醇可显著抑制肾癌786-O细胞的增殖并通过过度激活细胞自噬促进肾癌细胞凋亡。      

lncRNA NONHSAT113026在肾透明细胞癌中的表达及功能

目的:本研究旨在探讨lncRNA NONHSAT113026（lncRNA 026）在肾透明细胞癌（clear cell renal cell carcinoma,ccRCC）组织中的表达及对肾癌细胞增殖、迁移和侵袭的影响。方法:采用荧光定量聚合酶链反应（quantitative real-time polymerase chain reaction,qRT-PCR）分别在83例肾透明细胞癌和对应癌旁组织、肾癌细胞株786-O及ACHN中检测lncRNA 026的表达水平,并分析其表达水平与患者临床病理参数的相关性。通过慢病毒载体技术构建稳定过表达lncRNA 026的肾癌细胞786-O及ACHN,采用细胞计数试剂盒（cell counting kit-8,CCK-8）检测细胞增殖活性,Transwell小室检测细胞转移能力,蛋白印迹法（Western blot）分析PI3K/Akt信号通路中相关蛋白的表达情况。结果:与癌旁组织相比,lncRNA 026在ccRCC组织中的表达水平显著下调（P＜0.000 1）;在肾癌细胞786-O和ACHN中过表达lncRNA 026能显著抑制细胞的增殖、迁移和侵袭（P＜0.01）;Western blot结果显示,过表达lncRNA 026可下调PI3K/Akt信号通路中p-PI3K、p-Akt、GSK-3β的表达（P＜0.01）。结论:lncRNA 026在肾透明细胞癌组织中低表达,并且过表达lncRNA 026能显著降低肾癌细胞的增殖、侵袭和迁移能力,其作用机制可能与其抑制PI3K/Akt信号通路活化有关。提示lncRNA 026在肾癌中发挥了重要作用,有望成为治疗肾癌的潜在药物作用靶点。     

siRNA干扰FLOT2基因表达对肾细胞癌细胞增殖的影响

目的:探讨FLOT2在肾细胞癌细胞株中的表达,并观察小干扰RNA沉默FLOT2基因对肾细胞癌786-O细胞增殖能力的影响。方法:将化学合成的FLOT2的小干扰siRNA用脂质体 Lipofectamine2000转染至肾细胞癌786-O细胞;分别用RT-PCR、Western blot方法检测细胞转染前后FLOT2以及细胞增殖相关因子CCND1的表达。CCK-8与克隆形成实验检测其增殖情况。结果:转染FLOT2-siRNA的肾细胞癌786-O细胞,RT-PCR、Western blot检测显示FLOT2在基因及蛋白水平明显下调（P<0.05）;与对照组比较786-O细胞增殖能力明显下降（P<0.05）,细胞增殖相关因子CCND1明显下调（P<0.05）。结论:FLOT2在肾细胞癌细胞系中高表达,采用特异性FLOT2-siRNA 能够显著降低FLOT2和CCND1表达。干扰FLOT2表达能显著抑制786-O细胞的增殖,FLOT2基因可能参与了肾细胞癌的生物学行为。     

LNX1通过调控TIAM1/ERK信号通路抑制肾透明细胞癌细胞的增殖、侵袭和迁移

目的：探讨泛素链接酶LNX1(ligand of numb-protein X 1)对肾透明细胞癌细胞增殖、侵袭和迁移的影响及其可能的分子作用机制。方法：利用基因表达谱交互分析（Gene Expression Profiling Interactive Analysis,GEPIA）数据库分析LNX1 m RNA在肾透明细胞癌组织中的表达水平,及其与肾透明细胞癌患者预后的相关性。采用慢病毒感染的方法将携带特异性针对LNX1基因的shRNA(shLNX1)转入肾透明细胞癌细胞786-O和ACHN中;采用瞬时转染的方法将重组质粒flag-LNX1转入786-O和ACHN细胞中。分别采用CCK-8实验和细胞集落形成实验检测沉默或过表达LNX1对786-O和ACHN细胞增殖能力的影响,Transwell小室实验检测沉默或过表达LNX1对786-O和ACHN细胞侵袭和迁移能力的影响。采用生物信息学分析法筛选LNX1的下游靶基因,采用蛋白质印迹法检测沉默或过表达LNX1对T淋巴瘤侵袭和转移诱导蛋白1(T-lymphoma invasion and metastasis-inducing prote...     

反义survivin基因对786-O细胞的体外作用及其对表阿霉素诱导细胞凋亡作用的观察

的研究survivin反义寡核苷酸（ASODN）转染对肾透明细胞癌786-O细胞的survivin蛋白表达、细胞凋亡、增殖的影响,及其对表阿霉素诱导细胞凋亡的作用。方法设计并合成特异性靶向survivjn的ASODN及正义寡核苷酸（SODN）,将其转入786-O细胞。设空白对照组、脂质体对照组、SODN组和600nmol／L ASODN组,处理24h后收获各组细胞。透射电子显微镜观察细胞形态变化,免疫组化法检测各组细胞survivin表达情况,流式细胞术检测各组细胞增殖和凋亡指数。结果ASODN组的细胞呈典型凋亡的形态学改变,而对照组细胞生长良好;ASODN组细胞survivin表达减弱,凋亡指数明显升高（P〈0．05）;转染ASODN 24h后,表阿霉素诱导786-O细胞凋亡的作用明显增强。结论survivin ASODN能下调survivin蛋白表达,诱导肾透明细胞癌786-O细胞凋亡,抑制786-O细胞增殖,并增强表阿霉素诱导的细胞凋亡。     

干扰组蛋白赖氨酸甲基转移酶SET8对肾透明细胞癌786-O细胞增殖的影响

目的 细胞增殖是影响肾细胞癌进展的重要因素,本研究旨在探讨组蛋白赖氨酸甲基转移酶SET8在调节肾透明细胞癌786-O细胞增殖中的作用.方法 将体外培养肾透明细胞癌786-O细胞随机分为空白对照组、空质粒组和SET8-shRNA组3组.采用四甲基偶氮唑蓝(MTT)法检测3组细胞的增殖能力;RT-PCR和蛋白质印迹法检测3组细胞Survivin和Caspase-3的表达水平,分析干扰SET8对肾透明细胞癌786-O细胞增殖及相关基因和蛋白表达的影响.结果 shRNA-SET8可成功抑制786-O细胞中SET8蛋白的表达.MTT检测结果显示,0、12、24、36和48 h SET8-shRNA组增殖抑制率分别为(10.97±2.68)%、(20.38±7.12)%、(25.03±7.60)%、(30.35±1.97)%和(31.33±1.04)%,SET8-shRNA组786-O细胞的增殖能力较空白对照组和空质粒组明显降低,P<0.05.RT-PCR和蛋白质印迹法结果显示,SET8-shRNA组Survivin mRNA相对表达量为0.257±0.024,较空白对照组(1.046±0.041)和空质粒组(0.994±0.018)明显下降(P<0.05),其蛋白的相对表达量为0.834±0.072,较空白对照组(0.951±0.047)和空质粒组(1.203±0.092)明显下降,P<0.05;而Caspase-3 mRNA的相对表达量为0.921±0.072,较空白对照组(0.421±0.022)和空质粒组(0.439±0.007)明显上升(P<0.05),蛋白的相对表达量为1.188±0.022,较空白对照组(0.541±0.060)和空质粒组(0.617±0.028)明显上升,P<0.05.结论 干扰SET8可以有效抑制肾透明细胞癌786-O细胞SET8蛋白表达,从而抑制Survivin和上调Caspase-3的表达而抑制肾透明细胞癌786-O细胞增殖能力.     

桧木醇对肾癌786-O细胞增殖及相关细胞因子的影响

目的从体外水平研究桧木醇(hinokitio1)的抗肾癌作用,探讨其作用机制。方法采用MTT法和流式细胞术检测桧木醇对肾癌786-O细胞增殖的抑制作用和促凋亡作用的影响;采用Western-blot检测桧木醇对人肾癌786-O细胞Caspase-3剪切体、LC3和P62蛋白表达水平的影响。以3-MA验证其抗癌作用与自噬作用的关系。结果桧木醇对肾癌786-O细胞增殖有显著抑制作用,主要是通过激活Caspase途径对细胞凋亡进行诱导。同时桧木醇可以对肾癌786-O细胞进行诱导自噬发生,使LC3蛋白的表达量出现显著上调,而P62蛋白表达则显著下调。结论桧木醇能够显著抑制肾癌786-O细胞的增殖,而且可以通过过度激活细胞自噬促进肾癌细胞凋亡。     

小分子RNA干扰Notch1基因对肾癌Caki-1细胞增殖的影响

目的：探讨Notch1在肾透明细胞癌株中的表达,采用短片段RNA（small interferhtg RNA,siRNA）干扰技术沉默肾透明细胞癌中Notch1的表达,并观察其对Caki-1增殖的影响。方法：体外培养人正常肾小管上皮细胞HK-2及人肾癌细胞786—0和Caki-1。实时荧光定量PCR（RT—PCR）检测Notch1 mRNA的表达,蛋白印迹法检测Notch1蛋白的含量。特异性Notch1 siRNA干扰肾癌Caki-1细胞,用Lipofectamine2000转染48h后,实时荧光定量PCR和蛋白印迹法分别检测其中Notch1 mRNA、蛋白的表达变化,CCK-8与克隆形成实验检测其增殖情况。结果：Notch1在肾癌细胞株中的表达要明显高于正常肾小管上皮细胞,差异具有统计学意义（P〈0.05）。siRNA—Notch1干扰以后可以明显降低Caki-1细胞中Notch1 mRNA和蛋白的表达,差异具有统计学意义（P〈0.05）,同时检测Caki-1细胞增殖能力也随之下降（P〈0.05）。结论：Notch1在肾癌透明细胞癌中高表达,下调Notch1信号通路可以抑制肾细胞癌Caki—1的增殖,调节肾癌的生长。     

TET1对肾癌786-O细胞增殖的影响及其相关机制

目的:应用小干扰RNA(small interferingRNA,siRNA)沉默TET1(ten-eleven translocation1)基因表达,探索其对人肾癌786-O细胞增殖的影响及其可能的机制。方法:采用脂质体转染法将靶向TET1基因的TET1-siRNA转染人肾癌786-O细胞,实验设空白对照组、阴性对照组和干扰组(TET1-siRNA组)。分别采用实时荧光定量PCR和蛋白质印迹法检测转染TET1-siRNA后,786-O细胞中TET1、SUZ12(suppressor of zeste12)、EZH2(enhancer of zestehomolog 2)和EED(embryonic ectoderm development)mRNA及其蛋白的表达;MTT法和克隆形成实验检测786-O细胞的增殖活性和克隆形成率;FCM检测786-O细胞周期。结果:与空白对照组比较,转染TET1-siRNA后,786-O细胞中TET1 mRNA及其蛋白的表达水平分别下降了(85.13±0.03)%和(56.41±0.09)%,差异有统计学意义(P<0.05);786-O细胞的增殖受到抑制,克隆形成能力明显下降,G0/G1期细胞百分比明显增加(P<0.05);SUZ12 mRNA及其蛋白表达水平下调(P<0.05),EED和EZH2蛋白表达水平变化不明显(P>0.05)。结论:TET1-siRNA可显著下调肾癌786-O细胞中TET1蛋白的表达水平,明显抑制786-O细胞的增殖,并使细胞阻滞于G1期,其机制可能与TET1调控SUZ12的表达,影响其靶基因对PRC2(polycomb repressive complex 2)的招募有关。     

代谢综合征与肾透明细胞癌相关性研究

目的 代谢综合征(metabolic syndrome,MS)是一组临床症候群,MS及其相关组分与癌症发生发展及病理特征具有密切关系.本研究旨在分析MS及其相关组分与肾透明细胞癌(clear cell cenal cell carcinoma,CCRCC)分期、分级及肿瘤大小的相关性.方法 回顾性分析2013-01-01-2015-12-30于山西医科大学第一医院就诊且病理诊断为CCRCC的375例患者的临床资料,包括年龄、性别、身高、体质量、血压、空腹血糖、生化结果、病理分期分级和肿瘤大小等.计数资料采用x2检验,计量资料以(x)±s表示,组间比较采用t检验,多因素分析采用Logistic回归分析.结果 MS组56例患者,其中男性患者32例,女性患者24例;非MS组319例患者,其中男性患者206例,女性患者113例.男女患者比较,差异无统计学意义,P=0.287.MS组与非MS组相比,年龄、吸烟、饮酒等差异无统计学意义,P值分别为0.100、0.691和0.269;而BMI指数、收缩压、空腹血糖、TG、HDL-C等差异均有统计学意义,均P＜0.001.在病理特点方面,MS与非MS相比,CCRCC病理分期(P=0.018)、分级(P=0.026)及肿瘤大小(P=0.026),差异均有统计学意义.MS相关疾病与CCRCC分期分析,糖尿病(P＜0.001)、高血压(P=0.015)、血脂紊乱(P=0.006)与CCRCC的分期有关.结论 CCRCC合并MS者病理分期较高、分级较低、肿瘤更大,糖尿病、高血压和血脂紊乱都可增加CCRCC的病理分期.     

RhoB在肾透明细胞癌中的表达及临床意义

目的:探讨RhoB在肾透明细胞癌中的表达及临床意义.方法:采用实时定量PCR、Western blot和免疫组化方法检测RhoB在肾透明细胞癌组织中的表达情况.选取60例肾透明细胞癌组织标本,详细收集病人临床病理资料,通过免疫组化方法检测RhoB在肾透明细胞癌组织中的蛋白表达水平,并分析RhoB的蛋白水平与临床病理资料的关系.结果:与对应瘤旁肾组织相比,RhoB的mRNA及蛋白表达水平在肾透明细胞癌组织标本中明显降低;RhoB的蛋白表达水平在不同年龄、性别组间差异无统计学意义(P>0.05),而在肿瘤直径大小、T分期、临床分期和组织分级间差异有显著统计学差异(P<0.05).结论:RhoB的表达降低可能在肾透明细胞癌的肿瘤发生中发挥作用,且其表达下降可能与肾透明细胞癌的恶性进展有关.     

Origin of renal cell carcinomas

Cancer is a heritable disorder of somatic cells: environment and heredity are both important in the carcinogenic process. The primal force is the “two hits” of Knudson’s hypothesis, which has proved true for many tumours, including renal cell carcinoma. Knudson et al. [1, 2] recognised that familial forms of cancer might hold the key to the identification of important regulatory elements known as tumour-suppressor genes. Their observations (i.e., that retinoblastoma tend to be multifocal in familial cases and unifocal in sporadic presentation) led them to propose a two-hit theory of carcinogenesis. Furthermore, Knudson postulated that patients with the familial form of the cancer would be born with one mutant allele and that all cells in that organ or tissue would be at risk, accounting for early onset and the multifocal nature of the disease. In contrast, sporadic tumours would develop only if a mutation occurred in both alleles within the same cell, and, as each event would be expected to occur with low frequency, most tumours would develop late in life and in a unifocal manner [3, 4]. The kidney is affected in a variety of inherited cancer syndromes. For most of them, both the oncogene/tumour-suppressor gene involved and the respective germline mutations have been identified. Each of the inherited syndromes predisposes to distinct types of renal carcinoma. Families with hereditary predisposition to cancer continue to provide a unique opportunity for the identification and characterisation of genes involved in carcinogenesis. A surprising number of genetic syndromes predispose to the development of renal cell carcinoma, and genes associated with five of these syndromes have been already identified: VHL, MET, FH, BHD and HRPT2. Few cancers have as many different types of genetic predisposition as renal cancer, although to date only a small proportion of renal cell cancers can be explained by genetic predisposition. Supported by an unrestricted educational grant from Pfizer.     

MSCT多期扫描在肾脏乏脂肪血管平滑肌脂肪瘤与透明细胞癌鉴别诊断中的应用价值

目的 探讨肾脏乏脂肪血管平滑肌脂肪瘤(fpAML)和透明细胞癌(ccRCC)多排螺旋CT(MSCT)平扫及多期增强扫描表现,提高诊断准确率.方法 选取2015年1月至2019年12月江苏省中医院经手术病理确诊的fpAML和ccRCC患者各20例,测量肿瘤的短径和长径、CT平扫及多期增强的肿瘤CT值及邻近肾皮质的CT值,...     

Adjuvant therapy in renal cell carcinoma

Several drugs have demonstrated clinical activity in metastatic renal cell carcinoma (mRCC). The identification of key metabolic pathways has led to the development of novel targeted therapies which have drastically changed the treatment paradigm of mRCC. Moreover, immune-checkpoint inhibitors have recently shown significant activity in advanced disease. Despite these advancements, the role of adjuvant therapy in localized, non-metastatic RCC remains unclear. The utility of many of these agents in the adjuvant setting is currently being actively explored. In this review, we will summarize the main clinical trials investigating adjuvant therapy in renal cell carcinoma, focusing primarily on immunotherapy and targeted agents.     

MYC pathway is activated in clear cell renal cell carcinoma and essential for proliferation of clear cell renal cell carcinoma cells

Clear cell renal cell carcinoma (ccRCC) is the major and aggressive subtype of RCC. Previously, we identified 383 differentially expressed genes by analyzing full-length cDNA libraries of ccRCC and normal kidney tissues. In this study, we applied functional network analysis to the differentially expressed genes for identifying deregulated molecular pathways in ccRCC, and the results indicated that MYC showed a prominent role in the highest scoring network. The upregulation of MYC expression was validated in ccRCC tissues and cell lines. Furthermore, Knockdown of MYC expression by MYC-specific siRNA significantly inhibited the abilities of uncontrolled proliferation, anchorage-independent growth and arrested cell cycle in the G0/G1 phase in ccRCC cells. Moreover, we found that 37 differentially expressed genes were shown to be MYC-target genes, and the upregulation of the MYC-target genes BCL2, CCND1, PCNA, PGK1, and VEGFA were demonstrated. The expression of these MYC-target genes was significantly correlated with the expression of MYC in ccRCC tissues, and knockdown of MYC also suppressed the expression of these MYC-target genes in ccRCC cells. The recruitment of MYC to the promoter regions of BCL2, CCND1, PCNA, PGK1, and VEGFA was shown by Chromatin immunoprecipitation assay. These results suggest that MYC pathway is activated and plays an essential role in the proliferation of ccRCC cells.     

Nutrition and renal cell cancer

Epidemiologic evidence on the relation between nutrition and renal cell cancer is reviewed. Kidney cancer, comprising 1.7 percent of all malignant diseases diagnosed worldwide, shows about a 20-fold international variation in the incidence in men and 10-fold in women. This substantial variation indicates an important causal role of environmental factors. Renal cell (parenchymal) cancer (RCC) accounts for about 80 percent of all kidney cancers. While the etiology of RCC is incompletely understood, analytic epidemiologic studies provide consistent support for a positive association of obesity with risk of RCC; the dose-response observed supports a causal relationship. Only a few prospective studies, all of them limited in size, have been published, while ecologic and case-control studies suggest that diet may be important in the etiology of RCC. However, contradictory results and methodologic limitations in some casecontrol studies prevent definite conclusions concerning diet and RCC. A positive association of protein and fat intake, as well as their main food sources (meat, milk, fats), with risk of RCC-as suggested by ecologic studies—has no clear support in analytic epidemiologic studies. A protective effect of vegetables and fruits has been observed in most casecontrol studies, while the majority do not show an association between alcohol, coffee, and risk of RCC. Recent reports indicated an increased risk of RCC associated with consumption of fried/sautéed meat and low intakes of magnesium or vitamin E. An apparent positive association with total energy intake, perhaps due to bias, needs further investigation.This project was funded partially by the Swedish Cancer Society.     

Chemotherapy for non-clear-cell renal cell carcinoma

Clear-cell carcinoma is the most common histopathologic subtype of kidney tumors. Consequently, clinical trials for advanced-stage kidney cancer have focused on patients with clear-cell carcinoma and not on the less common subtypes, including papillary, chromophobe, collectingduct carcinoma, and sarcomatoid-variant tumors. Whereas immunotherapy has constituted the standard treatment for patients with clear-cell renal cell carcinoma (RCC), it does not appear to have activity in the management of patients with other histologic subtypes. Novel therapies, including those targeting the vascular endothelial growth factor pathway, have recently demonstrated significant activity in clear-cell RCC. Historically, chemotherapy has shown limited activity in advanced-stage RCC; however, clinical trials to date have failed to individualize treatment based on histologic subtype. In this article, we will review the literature and present our experience with the use of chemotherapy in patients with non-clear-cell kidney cancer by histologic subtype.     

Biomarkers in clear cell renal cell carcinoma

The treatment of advanced renal cell carcinoma (RCC) has evolved significantly following the identification of the von Hippel–Lindau (VHL) gene and the function of its protein, and subsequent development of antiangiogenic therapies. A series of clinical trials resulted in the approval of three new agents with significant activity in this disease. Additional studies are now underway to identify subsets of patients most likely to benefit. This article reviews the current therapy for advanced RCC and the development of biomarkers in RCC. This requires the identification of disease characteristics at a clinical, genetic and molecular level associated with response and/or surrogate measures of clinical benefit. Currently, a variety of prognostic factors (lactate dehydrogenase, performance status, disease-free interval, hemoglobin and calcium levels) are utilized to predict the survival of RCC patients. The use of validated biomarkers in either serum/plasma, urine or tissue could enhance this process, as well as define at the molecular and genetic levels, factors associated with response to therapy and/or the development of resistance. Examples include plasma VEGF levels, VHL gene mutation status and carbonic anhydrase IX levels in tumor tissue, among others. Validation of such biomarkers is crucial in order for them to be clinically useful.     

Measurement of progesterone receptor in human renal cell carcinoma and normal renal tissue

Progesterone receptor was measured in eight samples of renal cell carcinoma, nine samples of normal renal tissue, and one sample of melanoma tissue. Progesterone receptor was identified in all samples, with the exception of one renal cell carcinoma. Three patients, all with receptor-positive tumors, were treated with medroxyprogesterone acetate for metastatic disease. In one of these patients there was a partial objective response to treatment. Further research regarding progesterone receptor in renal cell carcinoma is indicated.