Rb-loss is associated with high malignancy in chondrosarcoma

Loss of function of the human retinoblastoma gene (Rb) is a frequent genetic abnormality in human malignancies and causes a disturbance in the cell cycle and loss of normal proliferation and differentiation. We studied the loss of heterozygosity (LOH) of the Rb gene in 31 formalin-fixed, paraffin-embedded cartilaginous tumors using polymerase chain reaction. The tumors were subdivided into 8 cases of dedifferentiated (DD) chondrosarcoma, 17 cases of conventional chondrosarcoma (nine grade 1, seven grade 2 and one grade 3), 4 enchondromas and 2 chondroblastomas. Both components of DD chondrosarcoma, the low-grade and anaplastic components, were separated by a microdissection approach. The genetic data were correlated with the expression of the Rb protein examined by Rb immunohistochemistry. We found Rb-LOH in one grade 3 chondrosarcoma, and in the anaplastic component in 7 of 8 cases of DD chondrosarcoma (89% of all high-grade chondrosarcomas). All tumors with Rb-LOH were immunohistochemically Rb-negative. The only case of DD chondrosarcoma negative for Rb-LOH in both components of the tumor also showed weak expression of the Rb protein in the anaplastic component. All benign cartilaginous tumors, low-grade chondrosarcomas and low-grade tumor components of DD chondrosarcomas were negative regarding Rb-LOH but positive in Rb immunohistostaining. We concluded that Rb-LOH predominantly occurs in high-grade chondrosarcomas. However, it is not a marker for identifying low-grade tumors with a tendency towards progression or local recurrence.     

高迁移率族蛋白1及晚期糖基化终产物受体在宫颈鳞状细胞癌中的表达及临床意义

目的 研究高迁移率族蛋白1(HMGB1)和晚期糖基化终产物受体(RAGE)在宫颈鳞状细胞癌(CSCC)组织中的表达及其与临床病理间的关系,探讨HMGB1/RAGE信号通路在CSCC转移中的作用.方法 采用荧光实时定量PCR、免疫组化及Western blot方法分别检测CSCC组织和正常宫颈鳞状上皮组织中HMGB1、RAGE基因及蛋白的表达.结果HMGB1在CSCC中的表达水平高于正常宫颈鳞状上皮组织(P＜0.05),其在CSCC组织中的表达与肿瘤分期、侵袭及转移明显相关(P＜0.05),而与肿瘤大小、分化程度无关(P＞0.05).RAGE mRNA的表达与CSCC转移明显相关(P＜0.05).转移组中的HMGB1 mRNA和RAGE mRNA表达水平呈显著相关性.结论 HMGB-1与CSCC发生、侵袭和转移相关,RAGE与CSCC转移相关,HMGB1/RAGE信号通路在CSCC转移过程中可能发挥重要作用.     

Expression and Clinical Significance of HMGB1 and RAGE in Cervical Carcinoma

OBJECTIVE To study the expression level and clinical significance of HMGB1 and RAGE in cervical squamous epithelial carcinoma. METHODS Real time quantitative polymerase chain reaction(qRT-PCR) was employed to examine the expression of HMGB1(high mobility group box protein1),and RAGE(receptor for advanced glycation endproducts)in 60 cervical squamous epithelial carcinomas(CSEC),their paraneoplastic tissues(PS)and 30 normal cervix tissues(NCS). RESULTS The expression of HMGB1 in the CSEC samples and PS was similar(P>0.05),but higher compared to NCS(P<0.05).Overexpression of HMGB1 in the CESC tissues was significantly correlated with the tumor (P<0.05),and the presence of metastasis(P<0.01),but not correlated with the tumor diameter or tumor grade.RAGE expression was not significantly different among these tissue types,and showed no significant correlation with the the tumor stage,diameter or grade.But there was a significant positive correlation between RAGE expression and CSEC metastasis. CONCLUSION The results suggest that HMGB1 may be related to the proliferation,progression and metastasis of CSEC.The relationship of HMGB1/RAGE may be of importance for CSEC metastasis.HMGB1 presents a new potential gene target for prevention and treatment of CSEC. Study of HMGB1/RAGE expression will offer an experimental foundation for understanding the pathogenesis of CSES.     

Co-expression of RAGE and HMGB1 is associated with cancer progression and poor patient outcome of prostate cancer

Receptor for advanced glycation end products (RAGE), along with its ligand high mobility group box 1 (HMGB1), is believed to play an important role in prostate cancer. The aim of this retrospective study was to investigate the expression of RAGE and HMGB1 and their clinical impact on prostate cancer progression and prognosis. The expression of RAGE and HMGB1 was assessed by immunohistochemistry in cancer lesions from 85 confirmed prostate cancer cases. We determined the potential association between the expression level of these two proteins and the clinicopathological features and overall patient survival. RAGE and HMGB1 were expressed in 78.8% (67/85) and 68.2% (58/85) cases of prostate cancer, respectively, and in the majority (54/85) of cases, these two proteins were co-expressed. There was a strong correlation between RAGE and HMGB1 expressions (P<0.001). The expression of RAGE, HMGB1 and their co-expression were all associated with advanced tumor clinical stage (P<0.05 for all). RAGE expression was also associated with the prostate specific antigen (PSA) level (P=0.014). However, neither the individual expression of those genes nor their co-expression was significantly related with age or Gleason score. The co-expression of RAGE and HMGB1 was associated with poor overall survival in patients with stage III and IV prostate cancer (P=0.047). These results suggest that the expression of RAGE and HMGB1 is associated with the progression and poor prognosis of prostate cancer. RAGE and HMGB1 could be new prognostic biomarkers for prostate cancer as well as molecular target for novel forms of therapies.     

Diminished levels of the soluble form of RAGE are related to poor survival in malignant melanoma

RAGE is a central driver of tumorigenesis by sustaining an inflammatory tumor microenvironment. This study links the soluble forms of RAGE (sRAGE and esRAGE) with clinical outcome of melanoma patients. Moreover, tissue expression of RAGE was analyzed using immunohistochemistry on two independent tissue microarrays (TMA) containing 35 or 257 primary melanomas, and 41 or 22 benign nevi, respectively. Serum concentrations of sRAGE and esRAGE were measured in 229 Stage III–IV patients using ELISA and plasma concentrations of sRAGE were analyzed in an independent second cohort with 173 samples of Stage I–IV patients. In this cohort, three well‐described SNPs in the RAGE gene were analyzed. RAGE protein expression was highly upregulated in primary melanomas compared to benign nevi in the two TMA (p < 0.001 and p = 0.005) as well as in sun‐exposed melanomas (p = 0.046). sRAGE and esRAGE were identified as prognostic markers for survival as diminished sRAGE (p = 0.034) and esRAGE (p = 0.012) serum levels correlated with poor overall survival (OS). Multivariate Cox regression analysis showed that diminished serum sRAGE was independently associated with poor survival (p = 0.009). Moreover, diminished sRAGE was strongly associated with impaired OS in the second cohort (p < 0.001). Multivariate Cox regression analysis including the investigated SNPs revealed an independent correlation of the two interacting promoter SNPs with impaired OS. In conclusion, the soluble forms of RAGE and variants in its genetic locus are prognostic markers for survival in melanoma patients with high risk for progression.     

RAGE和HMGB1在乳腺癌中表达及其临床意义

目的探讨乳腺癌组织中RAGE和HMGB1基因表达及其临床意义。方法各对50例早期、晚期乳腺癌组织及正常乳腺组织蜡块标本,运用real-time PCR技术检测RAGE和HMGB1基因的表达情况。并分析各基因表达与乳腺癌组织的分化程度、浸润深度、淋巴结转移、TNM分期之间的关系。结果 RAGE和HMGB1基因表达荧光实时定量PCR法上调分别为73%、79%。乳腺癌TNM分期、淋巴结转移与基因高表达有密切关系,RAGE基因表达与HMGB1表达呈正相关(P<0.05)。结论 RAGE和HMGB1基因表达对于乳腺癌早期诊断和预后分析有重要指导意义,根据其表达开展有选择性地基因治疗应有良好的应用前景。     

MYC amplification and polysomy 8 in chondrosarcoma: array comparative genomic hybridization, fluorescent in situ hybridization, and association with outcome.

PURPOSE: To identify recurrent regions of genomic gain or loss in chondrosarcoma in a clinically relevant and statistically valid fashion. Materials and METHODS: Array comparative genomic hybridization (CGH) results of 15 frozen tumor samples of high-grade chondrosarcoma for chromosome 8 are presented. A separate subset of 116 cartilaginous tumors with outcome data was used for validation. RESULTS: Array CGH identified gain at 8q24.12-q24.13, the region of the MYC (c-Myc) oncogene, as a frequent change in high-grade chondrosarcoma. In the validation arm of 116 cartilaginous tumors, MYC was frequently amplified in G2 (15%), G3 (20%), and dedifferentiated (21%) chondrosarcomas. No amplification was identified in samples of enchondroma and grade 1 chondrosarcoma. In samples without MYC amplification, polysomy 8 was a frequent finding in grade 1 (18%), grade 2 (31%), grade 3 (80%), and dedifferentiated (29%) chondrosarcomas, but was not found in any samples of enchondroma. MYC protein expression was identified in all samples with amplification, but was also frequent in the remaining samples without amplification or polysomy 8. Kaplan-Meier survival curves for overall survival showed a statistically significant difference for patients with MYC amplification or polysomy 8 (P = .034). Univariate analysis involving Cox proportional hazards models showed that grade (P = .003), polysomy 8 (P = .045), and MYC amplification (P = .053) correlated with shorter overall survival. By multivariate analysis, grade of chondrosarcoma (P = .026) was the only factor to reach statistical significance. CONCLUSION: MYC amplification and polysomy 8 can be used as markers of prognostic importance in chondrosarcoma. Molecular targeting of MYC expression may have therapeutic potential in the future for subsets of chondrosarcoma.     

HMGB1 as an autocrine stimulus in human T98G glioblastoma cells: role in cell growth and migration

HMGB1 (high mobility group box 1 protein) is a nuclear protein that can also act as an extracellular trigger of inflammation, proliferation and migration, mainly through RAGE (the receptor for advanced glycation end products); HMGB1–RAGE interactions have been found to be important in a number of cancers. We investigated whether HMGB1 is an autocrine factor in human glioma cells. Western blots showed HMGB1 and RAGE expression in human malignant glioma cell lines. HMGB1 induced a dose-dependent increase in cell proliferation, which was found to be RAGE-mediated and involved the MAPK/ERK pathway. Moreover, in a wounding model, it induced a significant increase in cell migration, and RAGE-dependent activation of Rac1 was crucial in giving the tumour cells a motile phenotype. The fact that blocking DNA replication with anti-mitotic agents did not reduce the distance migrated suggests the independence of the proliferative and migratory effects. We also found that glioma cells contain HMGB1 predominantly in the nucleus, and cannot secrete it constitutively or upon stimulation; however, necrotic glioma cells can release HMGB1 after it has translocated from the nucleus to cytosol. These findings provide the first evidence supporting the existence of HMGB1/RAGE signalling pathways in human glioblastoma cells, and suggest that HMGB1 may play an important role in the relationship between necrosis and malignancy in glioma tumours by acting as an autocrine factor that is capable of promoting the growth and migration of tumour cells. Rosaria Bassi and Paola Giussani contributed equally to this study.     

晚期糖基化终末产物受体在人子宫内膜癌中的表达及意义

目的:探讨晚期糖基化终末产物受体（RAGE）的表达是否与子宫内膜癌相关,并进一步分析其可能机制。方法:收集手术切除的子宫内膜癌石蜡标本,免疫组化方法检测内膜癌组织中RAGE的表达及微血管密度,利用图像分析系统分析RAGE蛋白在子宫内膜癌组织中的表达情况,以平均光密度值(IOD)代表RAGE的表达水平。用Pearson相关性分析来分析子宫内膜癌组织中RAGE与微血管密度的相关性。结果:与正常子宫内膜组织相比RAGE在内膜癌组织中呈显著高表达（P<0.01）,且在高度恶性的癌组织中的表达显著高于低度恶性癌组织（P<0.05）。子宫内膜癌患者癌组织中RAGE表达量与微血管密度呈正相关。结论:RAGE在子宫内膜癌中高表达,与内膜癌发生发展及血管新生相关。     

Expression of connective tissue growth factor in cartilaginous tumors

BACKGROUND: Connective tissue growth factor (CTGF) predominantly is expressed in hypertrophic chondrocytes and its specific receptors are demonstrated on chondrocytic cells. Therefore, CTGF may be involved in the proliferation and/or differentiation of cartilage cells. In the current study, CTGF expression was examined both in chondrosarcoma and enchondroma to clarify the relation between the expression of CTGF and the grade of malignancy. METHODS: The expression of CTGF and proliferating cell nuclear antigen (PCNA) were analyzed immunohistochemically in 34 cartilaginous tumor specimens. Eighteen tumors were determined to be chondrosarcoma including 8 Grade 1 tumors, 6 Grade 2 tumors, and 4 Grade 3 tumors. The percentage of CTGF positive and PCNA positive cells was quantified using at least 500 cells. RESULTS: CTGF was expressed in 70.1% of enchondroma cells, 84.0% of Grade 1 chondrosarcoma cells, 53.7% of Grade 2 tumor cells, and 26.8% of Grade 3 tumor cells (rho = -0.501; P = 0.0053). In chondrosarcoma cases, CTGF expression was correlated closely with tumor grade (rho = -0.920; P = 0.0001). There was a strong correlation between PCNA expression and tumor grade (rho = 0.907; P < 0.0001) and a strong negative correlation between CTGF and PCNA expression (rho = -0.493; P = 0.0061). In chondrosarcoma cases, patients with high expression of CTGF (>/= 30%) showed higher overall survival compared with those with low expression (< 30%) (P = 0.004). CONCLUSIONS: The current study revealed a correlation between the histologic grade of chondrosarcoma and prognosis, and the concomitant association between CTGF immunostaining and tumor grade and prognosis. Therefore, immunohistochemical staining with CTGF is a useful procedure for assessing the tumor grade and clinical course in patients with chondrosarcoma.     

P21WAF1/CIP1在卵巢上皮性肿瘤中的表达与临床意义的研究

目的 :探讨P2 1WAF1/CIP1在恶性卵巢上皮性肿瘤中的表达与临床意义。方法 :应用免疫组化S P检测P2 1WAF1/CIP1在 2 0例良性、10例交界性、5 5例恶性卵巢上皮性肿瘤中的表达 ,并分析P2 1WAF1/CIP1与恶性卵巢上皮性肿瘤临床病理指标的关系。结果 :P2 1WAF1/CIP1在良性、交界性与恶性卵巢上皮性肿瘤中的表达总阳性率及强阳性率均有显著性差异 (P =0 0 0 7,P强 =0 0 0 1) ,并随良性、交界性、恶性而降低表达 (P <0 0 1,P强 <0 0 0 1) ;与恶性卵巢上皮性肿瘤的临床分期、组织分化、淋巴转移、腹水及预后均有关 (P =0 0 11,0 0 0 0 ,0 0 14 ,0 0 11;P强 =0 0 37,0 0 0 3,0 0 16 ,0 0 0 2 ,P <0 0 5 )。结论 :P2 1WAF1/CIP1在恶性卵巢上皮性肿瘤的形成和发展中发挥重要作用 ,检测P2 1WAF1/CIP1蛋白表达情况对卵巢肿瘤的良恶性鉴别 ,判断预后 ,探索卵巢癌的发生过程 ,从而指导卵巢癌的基因治疗均有积极的意义。     

Overexpression of cathepsin B and urokinase plasminogen activator is associated with increased risk of recurrence and metastasis in patients with chondrosarcoma

BACKGROUND: Deregulation of the cellular protease network has been shown to be responsible for aggressive clinical behavior in several common human malignancies. In the current study, the authors evaluated the expression patterns of proteases in patients with chondrosarcoma of bone and correlated these patterns with clinical outcome. METHODS: The expression levels of urokinase plasminogen activator; matrix metalloproteinase types-1, -2, and -9; and cathepsins B and L were determined immunohistochemically in 114 cases of chondrosarcomas of bone and were correlated with their clinicopathologic parameters as well as with long term follow-up data. RESULTS: Overexpression of cathepsin B was associated with a high rate of local recurrence (P = 0.006) and a decreased recurrence free survival (P = 0.005). Overexpression of urokinase plasminogen activator was associated with an increased rate of metastasis (P = 0. 013), a decreased metastasis free survival (P = 0.016), and a decreased 5-year overall survival rate (P = 0.048). The univariate Cox model showed that tumor extension into soft tissue, high histologic grade, and overexpression of cathepsin B were predictors of adverse outcome. Multivariate analysis showed only overexpression of cathepsin B and tumor extension into soft tissue to be independent predictors of local recurrence. CONCLUSIONS: Overexpression of cathepsin B and urokinase plasminogen activator can be used to identify those patients with chondrosarcoma of bone who have an increased risk of local recurrence and distant metastases.     

Prognostic relevance of cell biologic and biochemical features in conventional chondrosarcomas

BACKGROUND: Conventional chondrosarcoma is the second most common malignant solid tumor of bone, and its management still poses a challenge for the orthopedic surgeon. Currently, tumor grade is the only parameter of prognostic significance besides stage and, possibly, resection margins. Additional independent prognostic markers therefore would be highly valuable for patient management. METHODS: In the current study, the authors evaluated biologic markers for various chondrocytic phenotypes by histochemical and immunohistochemical technology in a large series of clinically well defined cases of enchondromas and conventional chondrosarcomas, each with at least 5 years of clinical follow-up. RESULTS: The authors' results confirm the strong correlation between clinical behavior and cell differentiation as expressed by marker genes. The phenotypes of the tumor cells are the biologic substrate of the histopathologic appearance of the neoplasms and, thus, the biologic basis for classic tumor grading. Collagen Types II and X, as well as the proteoglycan aggrecan, suggest a mature neoplastic phenotype and good prognosis, i.e., low recurrence rate, rare metastasis, and long survival. Conversely, collagen Type I, together with cell spindling, indicates a transition to a more proliferative, so-called "dedifferentiated" phenotype, which clearly is associated with a poorer prognosis. The changes in cellular phenotypes are accompanied by changes in proliferative activity. Thus, low-grade neoplasms showing mainly mature and terminally differentiated (hypertrophic) chondrocytes display only scant proliferation whereas less differentiated chondrosarcomas with biologically dedifferentiated chondrocytes show significantly higher proliferative activity, a feature that is also highly correlated with prognosis. CONCLUSIONS: These data indicate that molecular markers are to a large extent the biologic basis of the conventional grading, rather than representing independent prognostic markers. The authors' results further indicate that COL1 has significant value in the distinction between enchondromas and low-grade chondrosarcomas including these that are histologically similar. Further understanding of chondrocytic phenotypes will be a promising way to provide new tumor markers for better understanding, diagnosis, and treatment of chondroid neoplasms.     

RAGE、sRAGE在结肠癌中的表达及其临床意义

目的：本研究通过检测结肠癌组织晚期糖基化终末产物受体（ the receptor for advanced glycation end product,RAGE）和血清可溶性RAGE（ soluble RAGE,sRAGE）的表达水平,探讨其在结肠癌发生、发展过程中的作用及其临床意义。方法：本研究纳入49例非糖尿病结肠癌（ I、II、III期）患者,以免疫组织化学评分法（ immunohistochemical score,IHS）评价结肠癌RAGE表达水平,以ELISA法检测患者外周血血清sRAGE浓度。结果：结肠癌组织RAGE表达水平（IHS为1.24±0.48）显著高于癌周组织（IHS为0.50±0.25）（P﹤0.05）, III期（IHS为1.42±0.51）显著性高于I期（IHS为1.01±0.35）（P﹤0.05）和II期（IHS为1.11±0.42）（P﹤0.05）;低分化结肠癌组织（IHS为1.58±0.49）显著高于中分化（IHS为1.08±0.45）（P﹤0.05）和高分化结肠癌组织（IHS为1.02±0.27）（P﹤0.05）。结肠癌患者血清sRAGE表达水平术前、术后分别为（344.77±68.06）ng/L、（265.43±76.85）ng/L,二者相比有显著性差异（ P ﹤0.05）,而且分别与健康志愿者 sRAGE （149.97±30.12）ng/L相比有显著性差异（ P﹤0.05）,但sRAGE表达水平与TNM分期、结肠癌组织分化程度没有关联。结肠癌组织RAGE高表达患者术前血清sRAGE浓度（415.02±85.08）ng/L高于中表达（334.26±44.56）ng/L和低表达（335.95±78.48）ng/L患者（P﹤0.05）。结论：结肠癌组织RAGE表达水平与分化程度呈负相关趋势,与TNM分期呈正相关趋势,作为术后诊断指标有一定的临床意义。血清sRAGE浓度与结肠癌组织RAGE表达水平呈正相关趋势,但与TNM分期和结肠癌组织分化程度没有关联,作为术前诊断指标仅能代表结肠癌风险,但无法考量结肠癌的恶性程度和进展情况。     

宫颈癌细胞高迁移率族警报素的分泌与肿瘤浸润淋巴细胞的相关性

目的:探讨高迁移率族蛋白B1和N1(high mobility group B1 and N1,HMGB1 and HMGN1)在宫颈癌肿瘤细胞中的表达情况与肿瘤浸润淋巴细胞(tumor infiltrating lymphocytes,TILs)的关系.方法:收集2012年4月至2015年9月在天津医科大学附属肿瘤医院就诊的100例早期宫颈癌患者的术后肿瘤组织蜡块标本,其中Ⅰ和Ⅱ期各50例.免疫组织化学方法检测肿瘤组织中HMGB1、HMGN1、CD3和CD8的表达.根据总体的染色强度将HMGB1和HMGN1分别分为高表达组和低表达组;根据有无细胞质表达分别分为有胞质表达组和无胞质表达组.分别比较高、低表达组间及有、无胞质表达组间肿瘤组织内CD3+和CD8+细胞数.结果:肿瘤组织内HMGB1和HMGN1的表达强度及有、无胞质表达与患者年龄、FIGO分期、病理分级均没有明显相关性(P＞0.05);HMGB1与HMGN1两者的胞质表达呈正相关关系(P＜0.05).HMGB1和HMGN1有胞质表达组的CD3+和CD8+细胞较无胞质表达组要高(P＜0.05).结论:宫颈癌细胞HMGB1和HMGN1的胞质表达与肿瘤组织内高水平TILs相关,这一发现有望为宫颈癌的免疫治疗提供新的策略.     

HMGB-1 induces cell motility and α5β1 integrin expression in human chondrosarcoma cells

Chondrosarcoma is a type of highly malignant tumor with a potent capacity to invade locally and cause distant metastasis. High mobility group box chromosomal protein 1 (HMGB)-1 is a widely studied, ubiquitous nuclear protein that is present in eukaryotic cells, and plays a crucial role in inflammatory response. However, the effects of HMGB-1 on human chondrosarcoma cells are largely unknown. In this study, we found that HMGB-1 increased the migration and the expression of α5β1 integrin in human chondrosarcoma cells. Transfection of cells with receptor for advanced glycation end products (RAGE) receptor siRNA reduced HMGB-1-induced cell migration and integrin expression. Activations of phosphatidylinositol 3-kinase (PI3K), Akt, and AP-1 pathways after HMGB-1 treatment were demonstrated, and HMGB-1-induced expression of integrin and migration activity was inhibited by the specific inhibitor and mutant of PI3K, Akt, and AP-1 cascades. Taken together, our results indicated that HMGB-1 enhances the migration of chondrosarcoma cells by increasing α5β1 integrin expression through the RAGE receptor/PI3K/Akt/c-Jun/AP-1 signal transduction pathway.