Sprouty2蛋白在肾透明细胞癌组织中的表达及意义

目的检测肾透明细胞癌及癌旁组织标本中Sprouty2蛋白的表达情况,分析其表达水平与肿瘤的临床进展及患者预后的相关性。方法采用免疫组化与Western blot方法对92例根治性肾切除肿瘤及癌旁组织标本以及8例非肿瘤肾组织中Sprouty2的表达进行检测,并结合临床病理学资料及预后进行统计学分析。结果肾癌中Sprouty2蛋白的表达较癌旁及非肿瘤肾组织中的表达显著减少,并且与肿瘤大小、有无转移、高分期、高分级有相关性(P均0.01)。Kaplan-Meier生存分析显示Sprouty2低表达组与高表达组之间差异有统计学意义(P0.01),单因素及多因素回归分析显示Sprouty2的表达水平可作为一个独立参数以评估肾癌患者的预后。结论 Sprouty2的低表达与肾癌的发生及进展相关,可能作为肾癌预测及预后评估的指标。     

EZH2在肾癌组织和肾癌细胞系中的表达及意义

目的：探讨EZH2蛋白在肾癌和肾癌细胞系中的表达情况及其与肾癌发生发展的关系。方法：应用Western blot检测12例肾癌和正常肾组织及两种细胞系ACHN和786—0中EZH2蛋白的表达,采用免疫组化法检测64例肾癌和12例正常肾组织中EZH2蛋白的表达情况,并分析与肾癌临床病理特征的关系。结果：West—ernblot检测结果显示,EZH2蛋白在正常肾小管上皮细胞系HK-2的表达极低,而肾癌细胞系786-0和ACHN的表达水平较高;正常肾组织EZH2蛋白表达阴性,肾癌组织均有不同程度的表达,差异有统计学意义（P〈0．05）。免疫组化结果示,肾癌和正常肾组织EZH2蛋白表达率分别为78．1％和16．7％,差异有统计学意义（P〈0．01）。EZH2蛋白表达率在局限性和局部进展性肾癌分别为63．9％和92．90A,差异有统计学意义（P〈0．01）;EZH2蛋白表达率在肾癌不同病理分级组和是否淋巴结转移组间差异有统计学意义（P％0．05）。结论：EZH2异常表达与肾癌的分期、分级和淋巴结转移有关,与肾癌的临床进展关系密切。     

白藜芦醇对肾癌细胞体外增殖与侵袭能力的影响

目的探讨白藜芦醇对肾癌786-O与ACHN细胞体外迁增殖与侵袭能力的影响及可能的机制。方法 MTT检测白藜芦醇对肾癌细胞增殖能力的影响,划痕实验检测白藜芦醇对786-O与ACHN细胞体外迁移能力的影响,Transwell实验检测白藜芦醇对786-O与ACHN细胞体外侵袭能力的影响,RT-PCR与Western bolt检测白藜芦醇对MMP-2与MMP-9蛋白表达水平的影响。结果白藜芦醇可抑制膀胱癌细胞体外生长能力,且呈剂量依赖性。30μmol/L白藜芦醇处理786-O与ACHN细胞24h后,划痕实验发现白藜芦醇可抑制786-O与ACHN细胞体外迁移能力,Transwell实验证实白藜芦醇可抑制肾癌细胞786-O与ACHN的体外侵袭能力。RT-PCR与Western bolt结果表明,白藜芦醇可从mRNA与蛋白水平抑制MMP-2与MMP-9的表达。结论白藜芦醇能抑制肾癌细胞体外增殖能力,可能通过下调MMP-2与MMP-9的表达而抑制肾癌细胞786-O与ACHN体外迁移与侵袭能力,有望成为治疗肾癌的新策略。     

自体LAK细胞和rIL－2治疗晚期肾癌的实验与临床研究

应用自体ＬＡＫ细胞和重组白细胞介素－２（ｒＩＬ－２）治疗２０例晚期肾癌患者。自患者周围血分离到的单个核细胞（ＰＢＭＣ）体外经ｒＩＬ－２短期培养，其ＮＫ、ＬＡＫ活性明显增强并于第５、７天达高峰。当这些ＬＡＫ细胞与ｒＩＬ－２过继回输给同一患者后，病人周围血ＮＫ、ＬＡＫ活性明显增加（Ｐ＜０．０ｌ），ＮＫ比率、ＩＬ－２受体表达明显增加（Ｐ＜０．０５），提示对肾癌患者的免疫调节作用。本组病人获部分缓解（ＰＲ）ｌ例，轻度缓解（ＭＲ）３例，平均缓解期５个月。毒副作用主要表现为发热、寒战，病人能耐受，表明ＬＡＫ／ｒＩＬ－２疗法是安全的方法。     

甲状旁腺素相关蛋白在肾癌细胞中的表达及调节作用

为研究甲状旁腺素相关蛋白在肾癌细胞中的表达及调节作用，采用免疫组化法对４１例肾癌标本进行染色，其中３９例（９５．１％）甲状旁腺素相关蛋白（ＰＴＨｒＰ）染色阳性，相比之下正常肾组织只有２０．６％染色阳性。ＰＴＨｒＰ染色结果与肿瘤大小、分期、复发和生存率无显著关系。肾癌细胞株Ｃａｋｉ１和Ｃａｋｉ２的培养液中加入不同浓度的ＰＴＨｒＰ受体竞争抑制物ＰＴＨｒＰ（７３４）后，两种细胞均受到与ＰＴＨｒＰ（７３４）浓度递度相应的抑制。上述结果表明ＰＴＨｒＰ在肾癌细胞中广泛表达，可能起到局部促进癌细胞生长的调节作用，该作用在体外实验中受ＰＴＨｒＰ（７３４）的抑制。     

NNMT在肾透明细胞癌的表达及对肾癌细胞侵袭能力的影响

目的:研究NNMT在肾透明细胞癌中的表达情况及对肾癌细胞侵袭能力的影响。方法:采用RT-PCR和Western blot方法检测正常肾小管上皮细胞株HKC、肾癌细胞株786-O及30例肾透明细胞癌组织、相应癌旁组织中NNMT的mRNA和蛋白的表达水平,并分析NNMT的mRNA水平与临床病理参数的关系。化学合成针对NNMT特异的siRNA序列,应用脂质体Lipofectamine 2000将其转染进786-O细胞中,利用RT-PCR和Western blot法检测NNMT在786-O细胞中的表达水平,用Transwell小室法检测肾癌细胞786-O侵袭能力的变化。结果:NNMT在肾癌细胞786-O中的mRNA和蛋白表达水平显著高于正常肾小管上皮细胞株HKC(P<0.001);肾透明细胞癌组织和对应的癌旁组织中NNMT的mRNA相对表达量分别为(1.582±0.2145)、(0.1269±0.04279),两组比较P<0.001。NNMT的mRNA水平与肿瘤大小、临床分期有关(P<0.05);Tran-swell法检测结果显示降低NNMT的表达后786-O细胞的侵袭能力明显下降。结论:NNMT在肾透明细胞癌组织和细胞中表达升高,可能在肾癌发生、发展过程中发挥重要作用。     

增殖细胞核抗原表达在肾细胞癌侵袭和转移性评估中的价值

侵袭和转移是影响肾细胞癌预后的重要因素 ,我们采用免疫组织化学技术检测 10 2例肾癌患者的癌细胞增殖细胞核抗原 (ＰＣＮＡ)表达 ,探讨其在肾癌侵袭和转移能力评估中的价值。1.资料和方法 :10 2例肾癌患者经根治术 ,取其肾癌标本 10 2份 ,根据是否存在浸润或转移 ,分为浸润转移组与非浸润转移组。远隔脏器转移 ,肾包膜外、肾外脏器浸润或肾、腔静脉癌栓依据术前影像学及术中探查、术后病理诊断 ;肾门淋巴结、镜下癌周组织及微静脉浸润由病理组织学检查 (图 1～ 3)确认 ,方法见李泉林等[1] 报道。应用标准过氧化物酶 链霉卵白素 (ＳＰ)免疫…     

小干扰RNA沉默血管内皮生长因子对ACHN肾癌细胞黏附、侵袭及迁移的影响

目的 观察小干扰RNA (siRNA)沉默血管内皮生长因子(VEGF)对ACHN肾细胞癌细胞生物学行为的影响.方法 化学合成针对VEGF的小干扰RNA,实验分4组,转染后收集细胞,通过蛋白印迹方法检测VEGF的表达,噻唑蓝(MTT)比色法、细胞黏附实验、划痕实验及Transwell法测定细胞的增殖、黏附、迁移及侵袭能力.结果 siRNA 1～2组VEGF表达水平明显低于空白对照组及阴性对照组;在24、48、72 h,siRNA 1～2的增殖抑制率均高于空白对照组及阴性对照组(P＜0.05),其中以siRNA2组最为显著;与空白对照组比较,siRNA 1～2组的细胞黏附数量明显下降[(81.5±3.1)％比(40.5±2.6)％、P＜0.05,(81.5±3.1)％比(22.5±2.4)％、P＜0.05],其中以siRNA2组最为明显;siRNA1～2组的细胞迁移数量明显下降(162±9比81±5,P＜0.05;162±9比38±4,P＜0.05);siRNA1～2组细胞侵袭能力明显下降(P＜0.05),且siRNA 2组的细胞侵袭能力明显低于siRNA 1组(P＜0.05).结论 VEGF基因在肾细胞癌细胞黏附、迁移和侵袭中发挥着重要作用;以靶向VEGF的siRNA转染肾细胞癌细胞,可抑制肾细胞癌细胞黏附、迁移和侵袭能力.     

自体LAK细胞和rIL—2治疗晚期肾癌的实验与临床研究

应用自体ＬＡＫ细胞和重组白细胞介素－２（ｒＩＬ－２）治疗２０例晚期肾癌患者。自患者周围血分离到的单个核细胞体外经ｒＩＬ－２短期培养，其ＮＫ、ＬＡＫ活性明显增强并于第５、７天达高峰。当这些ＬＡＫ细胞与ｒＩＬ－２过继回输给同一患者后，病人周围血ＮＫ、ＬＡＫ活性明显增加（Ｐ＜０．０１），ＮＫ比率、ＩＬ－２受体表达明显增加（Ｐ＜０．０５），提示对肾癌患者的免疫调节作用。本组病人获部分缓解（ＰＲ）１例，轻度     

姜黄素对人肾癌ACHN细胞放疗增敏作用的实验研究

目的研究姜黄素对人肾癌ACHN细胞放射增敏作用并探讨其作用机制。方法不同浓度姜黄素作用于ACHN人肾癌细胞24h后,MTT法检测姜黄素药物毒性;克隆形成实验观察其对放射敏感性的影响;流式细胞术检测姜黄素诱导细胞的凋亡率、细胞周期分布。结果姜黄素对人肾癌ACHN细胞有明显的抑制作用,可引起细胞的凋亡,且存在剂量和时间依赖;较低浓度的姜黄素（5μmol和10μmol）即可降低放射后ACHN细胞的克隆形成率,其放射增敏比分别为1．61及2．36;姜黄素及姜黄素联合辐射组的细胞周期阻滞在辐射敏感时相G2／M期;结论低剂量的姜黄素对人肾癌ACHN细胞有放射增敏作用,其机制可能与其引起细胞的凋亡增加,细胞周期阻滞有关。     

Relation of microvessel density with microvascular invasion, metastasis and prognosis in renal cell carcinoma

OBJECTIVE

To clarify the significance of microvessel density (MVD) in a retrospective investigation the relationship between the pattern of MVD (reflecting angiogenesis), and tumour stage, grade, size, and occurrence of microvessel invasion (MVI), metastasis, and cancer‐specific survival (CSS) in patients who had surgery for renal cell carcinoma (RCC).

PATIENTS AND METHODS

Vessels were labelled in sections of formalin‐fixed, paraffin‐embedded tissues from 54 RCCs by CD34 immunohistochemistry. The mean MVD, expressed as the number of vessels per 10 high‐power fields (HPF, ×400) were measured for each case. In addition, all pathological slides were reviewed for the presence and absence of MVI. The prognostic value of MVD and MVI was then evaluated, and correlated with the usual prognostic variables, tumour metastasis and CSS.

RESULTS

In a univariate analysis of CSS, the MDV tended to be lower as stage increased from pT1 to pT3, and as grade increased from G1 to G4, although it was statistically significant only for stage (P < 0.001 and 0.050, respectively). The mean MVD was higher in 42 nonmetastatic than in 12 metastatic tumours, and in 33 tumours associated with MVI than in 21 with no MVI (P < 0.001). The mean MVD was also lower and significantly different for 28 large than 26 small tumours (P = 0.005). The survival rate of patients with tumours that were small, low‐stage, of higher MVD, with no MVI and metastasis was significantly higher than that of patients with large, high‐stage, low MVD, with MVI and metastatic tumours (all P < 0.001). MVI was significantly more common with a decreasing trend in MVD and the presence of metastasis (Spearman rank correlation r_s = ?0.68, P = 0.01, and r_s = 0.39, P = 0.01, respectively). Independent prognostic factors in a multivariate analysis were: in all patients with RCC, tumour stage (P = 0.013) and metastasis (P = 0.028); in those with low MVD, MVI (P = 0.004) and metastases (P = 0.016); in those with no MVI, stage (P = 0.020); in those with MVI, MVD (P = 0.001); in those with no metastases, stage (P = 0.045); and in those with metastases, MVD (P < 0.001). No independent predictor was identified in patients with high MVD. In patients with no metastases there was a significantly shorter median CSS time in RCCs with low MVD and with MVI (P = 0.004 for both). Similarly, patients who had grade 3–4 tumours, vs those with lower MVD and with MVI, had a significantly shorter median CSS (P = 0.020 for MVD, and 0.01 for MVI).

CONCLUSIONS

This study suggested that MVD in RCC was inversely associated with MVI, tumour metastasis, patient survival and tumour diameter and stage, from the usual prognostic variables, but MVD was not an independent prognostic factor in multivariate analysis for all patients with RCC. Low MVD and the presence of MVI appears to be a marker for identifying patients with an adverse prognosis.     

Microscopic venous invasion in renal cell carcinoma as a predictor of recurrence after radical surgery

BACKGROUND: The objective of the present study was to investigate the significance of microscopic venous invasion (MVI) as a prognostic factor for patients with renal cell carcinoma (RCC) who underwent radical surgery. METHODS: The study included a total of 157 consecutive patients with non-metastatic RCC who underwent radical surgery between January 1986 and December 2002. The median follow-up period was 45 months (range 6-162 months). Microscopic venous invasion was defined by the presence of a cancer cell in blood vessels based on the examination of hematoxylin-eosin stained specimens. Other prognostic variables were assessed by multivariate analysis to determine whether there was a significant impact on cancer-specific and recurrence-free survivals. RESULTS: Microscopic venous invasion was found in 70 patients, and of this number, 17 (24.7%) developed a tumor recurrence and 12 (17.1%) died of cancer progression, while only six (6.9%) of the remaining 87 patients without MVI presented with disease-recurrence and three (3.5%) died of cancer. Among the factors examined, the presence of MVI was significantly associated with age, mode of detection, tumor size, pathological stage and tumor grade; however, only pathological stage was an independent predictor for disease-recurrence, and none of these factors were available to predict cancer-specific survival in multivariate analyses. In 120 patients with pT1 or pT2 disease, MVI was noted in 36 patients. In this subgroup, recurrence-free survival rates in patients with MVI were significantly lower than those in patients without MVI, and MVI was the only independent prognostic predictor for disease-recurrence in a multivariate analysis. CONCLUSION: Microscopic venous invasion is not an independent prognostic factor in patients with non-metastatic RCC who underwent radical surgery; however, it could be the only independent predictor of disease-recurrence after radical surgery for patients with pT1 or pT2 disease.     

Necrotising fasciitis of the thigh caused by duodenum invasion of renal cell carcinoma: A case report

Necrotising fasciitis, widespread necrosis of the skin, subcutaneous tissue, and superficial fascia, may be caused by many factors, among which underlying malignancy is observed rarely. We report a case with necrotising fasciitis of the lower extremity because of a duodenum to retroperitoneum fistula caused by renal cell carcinoma invasion. A 62‐year‐old male with newly diagnosed renal cell carcinoma was diagnosed with necrotising fasciitis at the end of 2 days in hospital. One day after debridement surgery, biliary contamination of dressings and tomography demonstrated fistulation from the duodenum to retroperitoneum and then to the right thigh because of renal tumour invasion. The second operation was performed to repair the duodenum. Intravenous antibiotics and hydration were maintained postoperatively. Although there was no surgical complication, the patient died because of respiratory collapse at the 12th day postoperatively. Renal cell carcinoma may invade the duodenum and, with retroperitoneal fistulation, may be the cause of necrotising fasciitis of the thigh. Laparotomy may be needed to control the origin of infection. However, necrotising fasciitis may be fatal in spite of aggressive treatment. The fasciitis should be diagnosed early to initiate timely aggressive treatment, and a possible endogenous source should be kept in mind.     

Cell proliferation, apoptosis, angiogenesis and growth rate of incidentally found renal cell carcinoma

BACKGROUND: Our previous study showed that the growth rate of incidentally found renal cell carcinoma (RCC) varied, and that the initial clinical and pathological features did not predict subsequent growth of the carcinoma. The objective of this study was to determine the relationships between cell proliferation, apoptosis, angiogenesis and the growth rates of these RCC. METHODS: We examined cell proliferation, apoptosis, and angiogenesis in 16 incidentally found cases of RCC. Cell proliferation was assessed by immunohistochemical staining with a Ki-67 antibody. Apoptosis was assessed by the terminal deoxynucleotidyl transferase (TdT) mediated deoxy-UTP biotin nick end labeling (TUNEL) technique. The Ki-67 labeling index (KI) and the apoptotic index (AI) were determined as the ratio of immunohistochemically positive cells per 1000 cancer cells. The KI/AI ratio was also determined. Angiogenesis was evaluated by CD34 immunostaining. Finally, we investigated the correlation between these parameters and the growth rate of primary lesions of incidentally found RCC. RESULTS: The KI ranged from 7 to 73 (median, 20), AI ranged from 6 to 171 (median, 26), and microvessel density (MVD) ranged from 21 to 673 (median, 265) for incidentally found RCC. Ki-67 labeling index, AI and MVD were not closely correlated to each other. Furthermore, these parameters were not associated with growth rates of incidentally found RCC. Only the KI/AI ratio was strongly correlated to the growth rate of incidentally found RCC (r = 0.709; P = 0.0083). CONCLUSION: Our results suggest that the balance between cell proliferation and apoptosis partly determines the growth rate of primary lesions of incidentally found RCC.