膀胱尿路上皮癌中PD-L1和PD-L2的表达及临床意义

目的 探讨膀胱尿路上皮癌(urothelial bladder cancer,UBC)中程序性死亡配体-1(programmed death-ligand 1,PD-L1)以及程序性死亡配体-2(programmed death-ligand 2,PD-L2)的表达及临床意义.方法 采用免疫组化法检测58例UBC组织中...     

The relation of beclin 1 and bcl-2 expressions in high grade prostatic intraepithelial neoplasia and prostate adenocarcinoma: A tissue microarray study

The aim of the present study was to evaluate the expressions of beclin 1 and bcl-2 in prostate cancer (PC) and high grade prostatic intraepithelial neoplasia (HGPIN), and to investigate their relationship with clinicopathological parameters. The study included 30 benign prostatic hyperplasia (BPH), 40 HGPIN and 106 primary PC cases. The expressions of beclin 1 and bcl-2 were assessed semiquantitatively based on both the percentage and intensity of positive staining cells. Beclin 1 was positive in 27 (90%) BPH, 37 (92.5%) HGPIN, and 90 (84.9%) PC cases (p > 0.05). Bcl-2 immunostaining was detected in 99 (93.4%) PC, 37 (92.5%) HGPIN, and 9 (30%) BPH cases (p < 0.0001). Regarding expression scores, beclin 1 was significantly lower in PC cases than in the HGPIN and BPH groups (p < 0.0001), and it was also negatively correlated with Gleason score (p = 0.004, r = −0.274). Bcl-2 expression score was significantly higher in PC than in the other groups (p < 0.0001), and also positively correlated with Gleason score (p < 0.0001, r = 0.425). Furthermore, a negative correlation was found between bcl-2 and beclin 1 expression scores in PC cases (p = 0.006, r = −0.265). Our results suggest an association between bcl-2 and beclin 1 expressions in malignant transformation of prostate tissue and also in regulating PC cell differentiation, progression and the aggressiveness of PC.     

RIN1在膀胱尿路上皮癌中的表达及临床特征

目的探讨膀胱尿路上皮癌中RINl蛋白的表达与临床病理学特征的关系。方法用sP免疫组化法检测手术切除的88例原发性膀胱尿路上皮癌标本和20例癌旁正常组织标本。所有组织用10％福尔马林固定。石蜡包埋,连续切片,厚度4μm。所有上述患者术前均未接受放疗,化疗。通过卡方检验,分析RINl表达与膀胱尿路上皮癌的临床病理学特点。结果免疫组化法发现,88例膀胱尿路上皮癌中有49例RINl表达升高,20例癌旁组织的RINl表达正常。RINl的表达与肿瘤的病理分级（P〈0．05）和临床分期（P〈0．05）呈正相关,与年龄、性别、肿瘤大小、是否多发则无相关性（P〉0．05）。结论RINI在膀胱尿路上皮癌中存在高表达,且在膀胱癌中发挥着重要作用。     

High expression of Notch ligand Jagged2 is associated with the metastasis and recurrence in urothelial carcinoma of bladder

Background: The Notch signaling pathway is closely related with human organ development and tumorgenisis. Jagged2 is among the most popular topic in Notch studies currently. Recent studies found its vital role in tumor metastasis in breast cancer; however, its expression profile and its prognostic value in urothelial carcinoma of bladder have not been investigated. Methods: Immunohistochemistry was used to detect the expression of Jagged2 in 120 bladder urothelial carcinoma. Moreover, the expression of Jagged2 was analyzed by Western blot in 60 bladder urothelial carcinoma and 20 normal epithelial tissues. MTT assay and flow cytometry and transwell assay were used to examine the proliferative and invasive ability of bladder cancer cells with the treatment of GSIXX (the inhibitor of Jagged2). Prognostic value of Jagged2 expression and its correlation with tumor metastasis and recurrence were evaluated, and the proliferative and invasive ability and cell cycle process of the bladder cancer cells were detected as well. Results: There was a significantly higher Jagged2 expressions in bladder urothelial carcinoma and highly invasive bladder T24 cells than those in bladder normal tissues and the superficial bladder BIU-87 cells. Jagged2 expression was positively correlated with histological grade, p T stage, recurrence, and metastasis. With the increasing concentration of GSIXX, we found that not only the cell proliferation and invasion activity decreased significantly, but also the cell cycle was blocked at G₂/M stage. Conclusions: Jagged2 expression status was closely correlated with important histopathologic characteristics (grades and stages) and the recurrence and metastasis of bladder urothelial carcinomas. Furthermore, Jagged2 played an important function on the bladder cancer cells’ proliferation by regulating the cancer cell cycle from G₁/S to G₂/M and probably promoted the invasion and metastasis of bladder cancer.     

Non-endothelial KDR/flk-1 expression is associated with increased survival of patients with urothelial bladder carcinomas

AIMS: To investigate the immunohistochemical expression of KDR/flk-1 in a series of 114 urothelial bladder carcinomas in relation to clinicopathological parameters, Ki67, p53 and Bcl-2 protein expression and patient survival. KDR/flk-1 is a high-affinity tyrosine kinase receptor for vascular endothelial growth factor (VEGF), on vascular endothelium. However, there is increasing evidence that KDR/flk-1 is also expressed by normal non-endothelial and tumour cells. METHODS AND RESULTS: Immunohistochemistry was performed on paraffin sections using monoclonal and polyclonal antibodies. Statistical analysis was univariate (chi2 log rank test) and multivariate (Cox's model). KDR/flk-1 expression was observed in the cytoplasm of cancerous cells in 68.4% of cases. No statistically significant associations were observed between KDR/flk-1 expression and grade or stage of urothelial carcinomas, Ki67, p53 or Bcl-2 expression. In contrast, widespread KDR/flk-1 expression in more than 50% of cancerous cells was associated with increased survival, on univariate and multivariate analysis (P = 0.0119 and P = 0.042, respectively). CONCLUSIONS: Although the biological significance of non-endothelial KDR/flk-1 expression has not yet been elucidated, its association with better patient survival may be related to the failure of non-endothelial KDR/flk-1 to mediate angiogenic and mitogenic effects.     

RIN1在人膀胱尿路上皮癌中的表达

目的研究RIN1在人膀胱尿路上皮癌中的表达。方法分别应用免疫蛋白印迹技术和实时定量PCR技术检测RIN1蛋白和RIN1 m RNA在人膀胱尿路上皮癌及癌旁组织中的表达。结果 Real-time PCR结果发现,在15对新鲜的膀胱尿路上皮癌及癌旁组织中,有12对的癌组织中RIN1表达高于癌旁组织。在膀胱尿路上皮癌组织中,RIN1的m RNA表达平均倍数为10.61±5.42,癌旁组织中,RIN1的m RNA平均倍数为4.31±1.77,P0.05。Western blot法结果认证了RIN1在膀胱尿路上皮癌组织中的表达是升高的。结论 RIN1在膀胱尿路上皮癌组织中的m RNA和蛋白的表达水平高于癌旁组织。     

自噬相关基因Beclin 1与肿瘤

自噬性细胞死亡是一种不同于凋亡、不依赖于caspase的程序性细胞死亡。研究发现自噬活性的变化、自噬性细胞死亡与恶性肿瘤的发生、发展有关。某些癌基因、抑癌基因参与了自噬活性的调节，其中对Beclin1研究较多，它作为自噬相关基因参与调节自噬活性，从而在肿瘤的发生、发展中发挥作用。现对Beclin1、自噬与肿瘤发生、发展关系的最新进展作一综述。     

PTEN与Beclin1在子宫内膜异位症的表达和临床意义

目的 考察PTEN与Beclin1在子宫内膜异位症的表达和临床意义.方法 采用免疫组化法测定40例子宫内膜异位症患者在位内膜、异位内膜和20例正常对照的PTEN与Beclin1表达水平.并考察不同病理资料与两者表达水平关联.结果 正常对照、在位内膜、异位内膜三者的PTEN与Beclin1表达水平均逐渐下降(P＜0.05).其中PTEN在正常对照、在位内膜和异位内膜的增生期、分泌期表达均无显著差异(P＞0.05).但Ⅲ-Ⅳ期表达率低于Ⅰ-Ⅱ期患者(P＜0.05).Beclin1在在位内膜和异位内膜的增生期、分泌期表达均无显著差异(P＞0.05),却在正常对照中具有显著差异(P＜0.05),同时Ⅲ-Ⅳ期表达率低于Ⅰ-Ⅱ期患者(P＜0.05).PTEN与Beclin1在子宫内膜异位症组织中表达水平正相关.结论 PTEN与Beclin1的低表达与子宫内膜异位症的发生和发展相关,但其具体作用机制值得进一步深入研究.     

TAp73和Beclin1在人骨肉瘤中表达及其临床意义

目的检测抑癌基因TAp73与自噬相关因子Beclin1在人骨肉瘤中的表达,探讨TAp73与肿瘤自噬的关系。方法采用免疫组织化学SP法检测TAp73、Beclin1在48例骨肉瘤患者的瘤体组织中的表达,结合患者的临床资料进行相关性分析。结果 TAp73在人骨肉瘤组织中的阳性率54.17%,Beclin1的阳性率为62.50%,TAp73阳性表达,其Beclin1表达程度较高(Z=-2.146,P=0.032)。在人骨肉瘤中TAp73的表达程度与Beclin1的表达程度呈正相关(r=0.470,P=0.001)。TAp73、Beclin1阳性表达的患者总体生存率优于阴性表达(80.8%vs 50.0%,P=0.000;80.0%vs 44.4%,P=0.001)。TAp73、Beclin1不同表达程度与患者的生存时间呈正相关(r=0.696,P=0.00;r=0.372,P=0.009)。结论在人骨肉瘤中TAp73可能与骨肉瘤中Beclin1依赖的肿瘤自噬活性上调有关。检测TAp73和Beclin1在人骨肉瘤中的表达水平有助于患者临床预后判断。     

Micropapillary variant of urothelial carcinoma of the urinary bladder; a clinicopathological and immunohistochemical study

AIMS: To investigate whether prognosis in micropapillary urothelial carcinoma is related to the proportion of the micropapillary component (MPC), and to identify the immunohistochemical features of MPC. METHODS AND RESULTS: This study presents a clinicopathological analysis of 20 patients with micropapillary urothelial carcinoma of the bladder with cystectomy specimens for evaluation. Tumours were stratified on the extent of MPC: focal, <10%; moderate, 10-50%; extensive, >50%; and this was correlated with tumour stage and prognosis. Sixteen males and four females were aged 56-81 years (mean 69 years). All cases had high-grade morphology in the micropapillary carcinoma and typical urothelial carcinoma. All cases with extensive MPC (n = 4) were of a high pathological stage (pT3 or pT4) and died of disease (DOD) or other causes. Eighty percent with moderate MPC (eight of 10 cases) were pT3 or pT4 and 50% DOD or are alive with disease. Eighty-four percent with focal MPC (five of six cases) were pT1 or pTa. In high-stage cases, the most invasive component was MPC. High-stage cases had an 85% risk of being advanced at presentation with micropapillary carcinoma. All pT2 or lower stage cases had micropapillary carcinoma on prior transurethral resections of bladder tumour (TURB). High-stage carcinomas had 30% and 54%, respectively, of surface MPC and urothelial carcinoma in situ, in comparison with 85% and 28% in lower stage carcinomas. Immunohistochemical staining was similarly positive in MPC and typical urothelial carcinoma with cytokeratin (CK)7, CK20, epithelial membrane antigen, carcinoembryonic antigen and cytokeratin 34betaE12. CA125 staining was seen only in MPC in 43% of cases. CONCLUSIONS: Micropapillary urothelial carcinoma is a high-grade carcinoma in which the prognosis is related to the proportion and location of the MPC. Cases with moderate or extensive MPC are at high risk of being advanced at presentation. Cases with <10% MPC and surface MPC have a high chance of detection at an early stage. The morphology and immunohistochemical profile of the MPC suggest that it is a form of glandular differentiation in urothelial carcinoma.     

大肠癌中Beclin1、LC3和mTOR的表达及意义

目的：探讨Beclin1、LC3和mTOR在大肠癌中的表达及意义。方法采用免疫组化EnVision法、Western blot和RT-PCR技术检测Beclin1、LC3和mTOR在大肠癌中的表达,并结合临床病理因素进行分析。结果 Beclin1、LC3和mTOR在大肠癌中的阳性率分别为90．50%、87．19%和46．28%,均明显高于癌旁组织( P<0．05),且LC3蛋白在中+低分化大肠癌中的表达(91．34%、89．58%)高于高分化大肠癌(77．61%),在有淋巴结转移组的表达(95．41%)高于无淋巴结转移组(80．45%), mTOR在有淋巴结转移组的表达(57．80%)高于无淋巴结转移组(35．33%)(P<0．05),Beclin1过表达与大肠癌分化程度和淋巴结转移均无关(P>0．05)。 LC3与Beclin1在大肠癌中表达呈正相关(rs =0．593,P<0．01),与mTOR表达呈负相关(rs =-0．165,P<0．01),Beclin1和mTOR表达之间无明显相关性(P>0．05)。 Kaplan-Meier生存分析显示,Beclin1和LC3均阳性、mTOR阴性、无淋巴结转移的大肠癌患者5年生存率高于Beclin1和LC3均阴性、mTOR阳性及有淋巴结转移的患者。多因素分析显示,LC3、mTOR和淋巴结转移是影响大肠癌预后的独立因素。 RT-PCR和Western blot检测结果显示：Beclin1、LC3和mTOR mRNA和蛋白表达水平均明显高于癌旁组织(P<0．05)。结论自噬相关基因Beclin1、LC3和mTOR的异常表达可能与大肠癌的发生、发展及预后相关;联合检测Beclin1、LC3和mTOR在大肠癌中的表达有助于评估进展程度和预后判断。     

Diagnostic algorithm for papillary urothelial tumors in the urinary bladder

Papillary urothelial neoplasms with deceptively bland cytology cannot be easily classified. We aimed to design a new algorithm that could differentiate between these neoplasms based on a scoring system. We proposed a new scoring system that enables to reproducibly diagnose non-invasive papillary urothelial tumors. In this system, each lesion was given individual scores from 0 to 3 for mitosis and cellular thickness, from 0 to 2 for cellular atypia, and an additional score for papillary fusion. These scores were combined to form a summed score allowing the tumors to be ranked as follows: 0–1 = UP, 2–4 = low malignant potential (LMP), 5–7 = low-grade transitional cell carcinoma (TCC), and 8–9 = high-grade TCC. In addition to the scoring system, ancillary studies of MIB and p53 indexes with CK20 expression pattern analyses were compared together with clinical parameters. The MIB index was strongly correlated with disease progression. Four of the 22 LMP patients (18.2%) had late recurrences, two of these four (9.1%) had progression to low-grade carcinoma. The MIB index for LMP patients was strongly associated with recurrence (recurrence vs. non-recurrence, 16.5 vs. 8.1, p < 0.001). The proposed scoring system could enhance the reproducibility to distinguish papillary urothelial neoplasms.     

c-myc, c-erbB-1 and c-erbB-2 expressions in urothelial carcinoma

The expression of c -myc , c- erb B-1 and c- erb B-2 In 24 cases of urothelial carcinoma by Southern and northern blot analysis, and immunohistochemistry was examined. The results were compared with the pathological grade and stage. We found elevated mRNA expressions of c- myc and c- erb B-1 In 19 and 11 of 21 cases, respectively, but there was no apparent amplification or rearrangement of these oncogenes in any of the cases examined. By immunohistochemistry using anti-epidermal growth factor receptor antibody, most of the cases showed positbe immunoreactivity on the cancer cell membranes, and cancers of higher pathological grade and stage showed more intense staining. By contrast, amplification of c- erb B-2 was detected in four of 24 cases, all of which were assigned to a high pathological grade (G3). Elevated c- erb B-2 mRNA levels appeared to correlate with the pathological grade of the cancers. Positive immunohistochemical reactions to c- erb B-2 were found in the cancer cell membranes in three of 24 cases, which were accompanied by amplification and eievated mRNA levels of c -erb B -2 . In conclusion, expressions of c- myc , c- erb B-1 and c- erb B-2 were all elevated in the majority of urothellal carcinomas, but the amplification was not universal.     

Gene rearrangement of bcl-1 and bcl-2 is confined to distinct subgroups of high-grade malignant B-cell lymphomas

The occurrence of bcl-1 and bcl-2 gene rearrangements was investigated in 37 cases of high-grade B-cell lymphomas. Bcl-2 rearrangement was detectable only in single cases of primary centroblastic lymphoma with a follicular growth pattern, whereas secondary centroblastic lymphomas evolving from a centroblastic–centrocytic lymphoma were positive in up to 60 per cent of the cases analysed. Bcl-1 rearrangement was found only in one case of immunoblastic B-cell lymphoma with a history of a pre-existing lymphoplasmacytoid immunocytoma. It is concluded that there may be a subgroup of centroblastic lymphomas with a biology similar to that of centroblastic–centrocytic lymphomas. The detection of bcl-1 rearrangement in high-grade lymphomas may indicate a secondary high-grade lymphoma.     

膀胱乳头状尿路上皮肿瘤中CD10和CK20的表达

目的探讨CD10与CK20在膀胱乳头状尿路上皮肿瘤中的表达及临床病理意义。方法以19例正常尿路上皮为对照,采用免疫组化法检测74例膀胱乳头状尿路上皮肿瘤中CK20和CD10染色分布模式和强度以及二者之间的关系。结果CD10与CK20的阳性率分别为51.4%和54.4%,两者的染色分布和强度差异无显著性(P0.05)。CK20的染色分布模式在尿路上皮乳头状瘤(urothelial papilloma,UP)和非浸润性乳头状尿路上皮癌(non-invasive papillary urothelial carcinoma,NIPUC)中差异有显著性(P0.05)。CD10与CK20的染色强度整体比较差异有统计学意义(P0.05),且UP、低度恶性潜能未定的乳头状尿路上皮肿瘤(papillary urothelial neoplasm of law malignant potential,PUNLMP)与NIPUC组间相比差异均有统计学意义(P0.05)。两者联合检测阳性率达74.3%,癌组织中阳性率为84.9%。结论 CD10与CK20在膀胱乳头状尿路上皮肿瘤中的表达极其相似,联合使用明显提高阳性率;对鉴别尿路上皮乳头状肿瘤表浅病变的类型以及该类肿瘤的良恶性有意义。     

Beclin 1 expression predicts favorable clinical outcome in patients with diffuse large B-cell lymphoma treated with R-CHOP

Beclin 1 plays a critical role not only in autophagy, but also in apoptosis and differentiation. The prognostic significance of beclin 1 in diffuse large B-cell lymphoma is largely unexplored. Immunohistochemical protein expression of beclin 1 in 118 tumor specimens from patients newly diagnosed with diffuse large B-cell lymphoma treated with rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone regimen from 2003 to 2007 was analyzed. Beclin 1 expression was observed in 101 (85.6%) patients. Low beclin 1 expression was related to B symptoms (P = .018), advanced Ann Arbor stage (P = .009), International Prognostic Index of 2 or higher (P = .049), elevated serum lactate dehydrogenase level (P = .037), and poor outcome using the revised International Prognostic Index model (P = .013). The complete remission rate after standard treatment was higher in patients with high beclin 1 expression than in those with low beclin 1 expression (77.5% versus 55.3%, P = .013). Beclin 1 expression was inversely associated with bcl-2 expression (P = .019) and positively associated with longer overall survival (P = .035) and progression-free survival (P = .013). In multivariate analysis, beclin 1 expression and the revised International Prognostic Index model independently predicted survival. The proposed clinicopathologic prognostic model including these 2 independent prognostic factors allowed classification of patients into 3 risk groups (P < .0001, respectively). Beclin 1 expression predicts superior clinical outcome in patients with diffuse large B-cell lymphoma treated with standard treatment. The novel prognostic model which divides patients with diffuse large B-cell lymphoma into different risk categories may help guide treatment planning.     

Expression of beclin 1 in primary salivary adenoid cystic carcinoma and its relation to Bcl-2 and p53 and prognosis

Beclin 1 plays a critical role in autophagy and functions as a haploinsufficient tumor suppressor. The expression and prognostic significance of beclin 1 in head and neck adenoid cystic carcinoma (ACC) are largely unexplored. Therefore, we investigated the expression of beclin 1, Bcl-2, and p53 in head and neck ACC tissue. Tissue samples from 35 cases (15 females, 20 males) of head and neck ACC were utilized for immunohistochemistry. Beclin 1 expression was observed in 32 cases (91.4%) and considered to be high in 15 cases (42.9%) and low in 20 cases (57.1%). Beclin 1 expression was significantly correlated with a histological growth pattern (P=0.046) and histological grade (P=0.037). Beclin 1 expression was inversely correlated with Bcl-2 expression (P=0.013) and significantly associated with overall survival (P=0.006). Bcl-2 and p53 expression were observed in 21 cases (60.0%) and 16 cases (45.7%). Bcl-2 expression was significantly correlated with perineural invasion (P=0.041) and not associated with overall survival (P=0.053). p53 expression was directly correlated with beclin 1 expression (P=0.044). Our results indicated that beclin 1 may be a novel, promising prognostic factor for clinical outcome in head and neck ACC patients and may play a part in the development of head and neck ACC by interacting with Bcl-2 and p53.     

自噬基因Beclin 1 shRNA表达质粒的构建及其下调凋亡因子caspase-9效应的研究

构建自噬基因Beclin 1的小发夹RNA(shRNA)真核表达质粒,脂质体包裹后体外转染人宫颈癌HeLa细胞株,通过荧光定量PCR(RT-PCR)和Western blot检测其对HeLa细胞自噬基因Beclin 1 mRNA及蛋白表达的影响,并检测细胞增殖、细胞周期、细胞凋亡情况以及凋亡因子caspase-9 mRNA及蛋白表达的变化。结果表明shRNA真核表达载体可以使HeLa细胞中自噬基因Beclin 1的mRNA及其蛋白含量降低,转染后细胞生长增殖速度加快,凋亡率降低,并伴有caspase-9 mRNA及蛋白量的显著下调。因此,Beclin 1不仅与自噬调控通路有关,而且可以调控凋亡的发生,同时参与两种程序性细胞死亡的过程。     

Expression and function analysis of indoleamine 2 and 3-dioxygenase in bladder urothelial carcinoma

Indoleamine 2, 3-dioxygenase (IDO) is a rate-limiting enzyme for tryptophan metabolism inducing immune tolerance of tumors. The purpose of this study is to investigate IDO expression and its prognostic significance in bladder urothelial carcinoma (BUC). In this study, immunohistochemical staining for IDO expression in BUC tissues (n = 84) and normal bladder tissues (n = 22) was performed. The mRNA expression levels of IDO in BUC and normal bladder were analyzed by quantitative RT-PCR. Survival analysis was performed for the correlation of IDO expression and clinicopathological factors with disease-free survival. Positive expression of IDO was found in 48 of 84 cases in BUC tissues and was significantly correlated with histological classification, histological grade and TNM stage. While IDO expression in normal bladder tissues was expressed in only 4 of 22 (18.2%) cases. Moreover, IDO mRNA levels of BUC were significantly higher than that of normal bladder. We also found that IDO, histological grade and TNM stage were closely associated with DFS. These results indicated that IDO was related to the progression of BUC and might be one of the crucial prognostic factors for BUC.