Reduced expression of chemokine (C-C motif) ligand-2 (CCL2) in ovarian adenocarcinoma

Chemokine (C-C motif) ligand-2 (CCL2) is a chemoattractant and activator of macrophages and is a key determinant of the macrophage infiltrate into tumours. We demonstrate here that CCL2 is expressed in normal human ovarian surface epithelium (HOSE) cells and is silenced in most ovarian cancer cell lines, and silenced or downregulated in the majority of primary ovarian adenocarcinomas. Analysis of the CCL2 locus at 17q11.2-q12 showed loss of heterozygosity (LOH) in 70% of primary tumours, and this was significantly more common in tumours of advanced stage or grade. However, we did not detect any mutations in the CCL2 coding sequence in 94 primary ovarian adenocarcinomas. These data support the hypothesis that CCL2 may play a role in the pathobiology of ovarian cancers, but additional studies will be required to evaluate this possibility.     

Cancer testis antigens expression in mesothelioma: role of DNA methylation and bioimmunotherapeutic implications

Recent evidences suggest that malignant mesothelioma may be sensitive to immunotherapy; however, little is known about malignant mesothelioma-associated tumour antigens. Focusing on cancer/testis antigens, the expression of well-characterised immunogenic tumour-associated antigens was investigated in malignant mesothelioma cells. At variance with MAGE-4 and NY-ESO-1, malignant mesothelioma cells frequently expressed MAGE-1, -2 and -3, GAGE 1-2, GAGE 1-6, SSX-2 and SSX 1-5, and distinct malignant mesothelioma cells concomitantly expressed at least four cancer/testis antigens. Additionally, the tumour-associated antigens RAGE-1 was expressed at high levels in both benign and malignant mesothelial cells. Lastly, treatment with the DNA hypomethylating agent 5-aza-2'-deoxycytidine induced and up-regulated the expression of the cancer/testis antigen examined in malignant mesothelioma cells. Overall, these findings strongly suggest that cancer/testis antigens-based immunotherapy may represent a suitable therapeutic approach to malignant mesothelioma, and foresee the clinical use of 5-aza-2'-deoxycytidine to design new chemo-immunotherapeutic strategies in malignant mesothelioma patients.     

ICAM-1在肿瘤中的研究进展

细胞间黏附分子(Intercellular adhesion molecule-1,ICAM-1)又称CD54,是免疫球蛋白超家族的一种跨膜糖蛋白,参与肿瘤细胞免疫调节、血管生成、侵袭和远处转移。国内外诸多研究证明ICAM-1在多种恶性肿瘤中异常高表达,促进了肿瘤的发生、发展并影响其预后,同时对预测个别肿瘤放化疗敏感性有一定的作用。因此ICAM-1可能为恶性肿瘤的诊断和治疗提供参考依据。本文就近年来ICAM-1在恶性肿瘤中的研究做一综述。     

Amphiregulin enhances intercellular adhesion molecule-1 expression and promotes tumor metastasis in human osteosarcoma

Osteosarcoma is a common, high malignant, and metastatic bone cancer. Amphiregulin (AREG) has been associated with cancer cellular activities. However, the effect of AREG on metastasis activity in human osteosarcoma cells has yet to be determined. We determined that AREG increases the expression of intercellular adhesion molecule-1 (ICAM-1) through PI3K/Akt signaling pathway via its interaction with the epidermal growth factor receptor, thus resulting in the enhanced cell migration of osteosarcoma. Furthermore, AREG stimulation increased the association of NF-κB to ICAM-1 promoter which then up-regulated ICAM-1 expression. Finally, we observed that shRNA silencing of AREG decreased osteosarcoma metastasis in vivo. Our findings revealed a relationship between osteosarcoma metastatic potential and AREG expression and the modulating effect of AREG on ICAM-1 expression.     

A randomised,phase II trial of the DNA-hypomethylating agent 5-aza-2′-deoxycytidine (decitabine) in combination with carboplatin vs carboplatin alone in patients with recurrent,partially platinum-sensitive ovarian cancer

Background:

Our previous laboratory and clinical data suggested that one mechanism underlying the development of platinum resistance in ovarian cancer is the acquisition of DNA methylation. We therefore tested the hypothesis that the DNA hypomethylating agent 5-aza-2′-deoxycytodine (decitabine) can reverse resistance to carboplatin in women with relapsed ovarian cancer.

Methods:

Patients progressing 6–12 months after previous platinum therapy were randomised to decitabine on day 1 and carboplatin (AUC 6) on day 8, every 28 days or carboplatin alone. The primary objective was response rate in patients with methylated hMLH1 tumour DNA in plasma.

Results:

After a pre-defined interim analysis, the study closed due to lack of efficacy and poor treatment deliverability in 15 patients treated with the combination. Responses by GCIG criteria were 9 out of 14 vs 3 out of 15 and by RECIST were 6 out of 13 vs 1 out of 12 for carboplatin and carboplatin/decitabine, respectively. Grade 3/4 neutropenia was more common with the combination (60% vs 15.4%) as was G2/3 carboplatin hypersensitivity (47% vs 21%).

Conclusions:

With this schedule, the addition of decitabine appears to reduce rather than increase the efficacy of carboplatin in partially platinum-sensitive ovarian cancer and is difficult to deliver. Patient-selection strategies, different schedules and other demethylating agents should be considered in future combination studies.     

Osteopontin regulates human glioma cell invasiveness and tumor growth in mice

Human malignant glioma cells are characterized by local invasion. In the present study, we investigated the role of osteopontin (OPN) in the invasiveness of human glioma cells isolated from grade IV tumors. We found that the expression levels of OPN in these cell lines paralleled matrix metalloproteinase-2 (MMP-2) expression and cell invasiveness potential. When U87MG glioma cells (with a high-OPN expression level) were stably transformed with specific small hairpin RNA to knock down OPN expression, MMP-2 secretion, cell invasiveness, and tumor growth in implanted brains were dramatically reduced. Conversely, forced expression of OPN in GBM-SKH glioma cells (which expressed OPN at a low level) increased MMP-2 secretion, enhanced cell invasiveness, and increased tumor growth in a rodent xenograft model. Expression of OPN was associated with increased expression of vimentin and decreased expression of glial fibrillary acidic protein. Treatment of glioma cells with 5-aza-2′-deoxycytidine (5-aza-dC) suppressed OPN expression in a concentration-dependent manner. Suppression of OPN expression by 5-aza-dC was associated with reductions in MMP-2 secretion, vimentin expression, cell invasion, intravasation, and tumor growth. These data suggest that OPN may play important roles in regulating cell invasion in glioma cells and that 5-aza-dC may serve as a therapeutic agent for human gliomas.     

Inflammation-associated gene expression is altered between normal human ovarian surface epithelial cells and cell lines derived from ovarian adenocarcinomas

Ovulation is believed to contribute to the development of ovarian cancers that derive from the ovarian surface epithelium (OSE). The process of ovulation is synonymous with inflammation and inflammatory cytokines such as interleukin-1alpha (IL-1alpha) have recently been shown to induce both inflammatory and anti-inflammatory responses in human OSE (HOSE) cells. In this study we directly compared levels of IL-1alpha-induced gene expression by analysing the levels of 11beta-hydroxysteroid dehydrogenase (11betaHSD) types 1 (11betaHSD-1) and 2 (11betaHSD-2), cyclooxygenase-2 (COX-2), IL-1 receptor (IL-1R) and glucocorticoid receptor alpha (GRalpha) mRNA between normal HOSE cells and cell lines derived from poorly differentiated (SKOV-3, BG-1, PEO-4) and well-differentiated (PEO-14) ovarian adenocarcinoma. In HOSE cell cultures, and to a lesser extent PEO-14 cells, the basal mRNA levels of COX-2 and 11betaHSD-1 were relatively high and further shown to be induced in response to IL-1alpha (for HOSE cells; >20-fold, P<0.05 and PEO-14 cells; >3fold, P<0.05). However, whereas HOSE cells expressed a low level of 11betaHSD-2 mRNA that was only mildly responsive to IL-1alpha (1.3-fold, P<0.001), all cell lines exhibited a higher basal level of 11betaHSD-2 mRNA that was in some cases further stimulated in PEO-4 cells (five-fold; P<0.05) or suppressed in SKOV-3 cells (two-fold; P<0.01) in response to IL-1alpha. All cells tested expressed IL-1R and, with the exception of BG-1, GRalpha. These results indicate that cell lines derived from ovarian cancers have lost the ability to respond normally to inflammatory cytokines such as IL-1alpha. The finding that normal OSE cells, in contrast to cell lines derived from patients with ovarian adenocarcinoma, abundantly express 11betaHSD-1 mRNA but are essentially devoid of 11betaHSD-2 mRNA supports the concept that the pattern of 11betaHSD isoform gene expression is a defining feature of neoplastic cellular transformation, which might have particular relevance to the ovary.     

血清细胞间黏附分子-1 在胰腺癌诊断和预后评估中的价值

[摘要] 目的：探讨血清细胞间黏附分子-1（intercellular adhesion molecule-1,ICAM-1）在胰腺癌诊断和预后评估中的价值。方法：选取2015 年4 月至2017 年12 月在湖北省肿瘤医院肝胆胰外科就诊的胰腺癌患者80 例（胰腺癌组）、胰腺良性疾病患者40例（良性疾病组）及同期健康体检者30 例（对照组）。分别检测3 组人群血清ICAM-1 和CA19-9 水平;采用受试者工作特征曲线（ROC）分析ICAM-1 对胰腺癌的诊断特性,采用COX回归模型分析血清ICAM-1 与胰腺癌患者预后是否独立相关。结果：胰腺癌组ICAM-1 和CA19-9 水平明显高于良性疾病组和对照组（均P<0.01）,良性疾病组CA19-9 水平明显高于对照组（P<0.01）。血清ICAM-1、CA19-9 以及两者联合的曲线下面积（AUC）为0.732（95%CI：0.658~0.807,P=0.000）、0.691（95%CI:0.620~0.762, P=0.000）、0.747（95%CI ：0.674~0.821,P=0.000）;ICAM-1与CA19-9之间呈显著正相关（r=0.472,P=0.000）。血清ICAM-1<2 308 U/ml患者的生存时间明显长于≥2 308 U/ml 的患者（χ2=28.357,P=0.000）;ICAM-17≥2 308 U/ml 是患者预后的独立影响因子,其OR为3.08（2.14~7.23）。结论：血清ICAM-1 有助于胰腺癌的早期诊断和预后评估。     

Mutation and expression analysis of LZTS1 in ovarian cancer

LZTS1 has been shown to have tumour suppressor activities against prostate and breast cancer and is located within a region of frequent loss of heterozygosity (LOH) at 8p22 in ovarian cancer. We have analysed the expression of LZTS1 in ovarian cancer and found no evidence of loss of expression relative to normal ovarian surface epithelial cells. We have also analysed the coding region of the LZTS1 gene in 87 primary ovarian adenocarcinomas by DHPLC and detected a single silent somatic mutation. These data indicate that LZTS1 is not the target of LOH at 8p22 in ovarian cancer.     

Identification of novel hypermethylated genes and demethylating effect of vincristine in colorectal cancer

Background

Colorectal cancer (CRC) arises as a consequence of genetic events such as gene mutation and epigenetic alteration. The aim of this study was to identify new hypermethylated candidate genes and methylation-based therapeutic targets using vincristine in CRC.

Methods

We analyzed the methylation status of 27,578 CpG sites spanning more than 14,000 genes in CRC tissues compared with adjacent normal tissues and normal colon tissues using Illumina bead chip array. Twenty-one hypermethylated genes and 18 CpG island methylator phenotype markers were selected as candidate genes. The methylation status of 39 genes was validated by quantitative methylation-specific polymerase chain reaction in CRC tissues, adjacent normal tissues, normal colon cells, and three CRC cell lines. Of these, 29 hypermethylated candidate genes were investigated using the demethylating effects of 5-aza-2′-deoxycytidine (5-aza-dC) and vincristine in CRC cells.

Results

Thirty-two out of 39 genes were hypermethylated in CRC tissues compared with adjacent normal tissues. Vincristine induced demethylation of methylated genes in CRC cells to the same extent as 5-aza-dC. The mRNA expression of AKR1B1, CHST10, ELOVL4, FLI1, SOX5, STK33, and ZNF304 was restored by treatment with 5-aza-dC and vincristine.

Conclusion

These results suggest that these novel hypermethylated genes AKR1B1, CHST10, ELOVL4, SOX5, STK33, and ZNF304 may be potential methylation biomarkers and therapeutic targets of vincristine in CRC.     

Expression of intercellular adhesion molecule-1 in hepatocellular carcinoma

The expression of intercellular adhesion molecule-1 (ICAM-1) was investigated in frozen sections obtained from 40 resected liver specimens of patients with hepatocellular carcinoma using immunoperoxidase techniques and immunoelectron microscopy. ICAM-1 was expressed in 80% of the HCC specimens on the membrane of cancer cells. In noncancerous regions characterized by cirrhosis in 28 cases and chronic hepatitis in 12 cases, ICAM-1 was rarely expressed on hepatocytes but was expressed mainly on the endothelium of portal vessels and sinusoidal lining cells. These results suggest that expression of ICAM-1 in hepatocellular carcinoma may be induced by malignant transformation of hepatocytes. © 1993 Wiley-Liss, Inc.     

Identification of differentially expressed genes according to chemosensitivity in advanced ovarian serous adenocarcinomas: expression of GRIA2 predicts better survival

Background:

The purpose of this study was to identify genes that are differentially expressed in chemosensitive serous papillary ovarian carcinomas relative to those expressed in chemoresistant tumours.

Methods:

To identify novel candidate biomarkers, differences in gene expression were analysed in 26 stage IIIC/IV serous ovarian adenocarcinomas (12 chemosensitive tumours and 14 chemoresistant tumours). We subsequently investigated the immunohistochemical expression of GRIA2 in 48 independent sets of advanced ovarian serous carcinomas.

Results:

Microarray analysis revealed a total of 57 genes that were differentially expressed in chemoresistant and chemosensitive tumours. Of the 57 genes, 39 genes were upregulated and 18 genes were downregulated in chemosensitive tumours. Five differentially expressed genes (CD36, LIFR, CHL1, GRIA2, and FCGBP) were validated by quantitative real-time PCR. The expression of GRIA2 was validated at the protein level by immunohistochemistry, and patients with GRIA2 expression showed a longer progression-free and overall survival (P=0.051 and P=0.031 respectively).

Conclusions:

We found 57 differentially expressed genes to distinguish between chemosensitive and chemoresistant tumours. We also demonstrated that the expression of GRIA2 among the differentially expressed genes provides better prognosis of patients with advanced serous papillary ovarian adenocarcinoma.     

宫颈癌组织ICAM-1和VEGF表达与放疗敏感相关性研究

目的：研究细胞间黏附分子-1（intercellulareelladhesionmolecule-1,ICAM-1）和血管内皮生长因子（vascularendo—thalialgrowthfactor,VEGF）在宫颈鳞癌组织中的表达及其与放疗敏感性的相关性。方法：选取2007—07—01—2008—06—30在南通市肿瘤医院妇瘤科住院行根治性放疗的60例宫颈鳞癌患者。采用Realtime—PCR技术检测ICAM-1mRNA和VEGFmRNA在宫颈鳞癌组织中的表达,分析不同放疗敏感性患者IcAM-1InRNA和VEGFITIRNA表达水平差异,探讨ICAM-1rflRNA和VEGFmRNA之间的相关性。结果：宫颈鳞癌组织放疗前后ICAM-11TIRNA表达水平分别为124．57±58．64和47．38±39．27,差异有统计学意义,p=0．008;VEGFITIRNA的表达水平分别为152．75±66．92和73．46士54．41,差异有统计学意义,p=0．023;ICAM-1mRNA和VEGFmRNA的表达在放疗抵抗组和放疗敏感组中差异均有统计学意义,P值分别为0．027和0．009;近期疗效中,肿瘤消失组中ICAM-1mRNA和VEGFmRNA表达存在-定相关性,r=0．468,p=0．016;肿瘤未控组中IcAM-1nlRNA和VEGFmRNA表达存在-定相关性,r=0．693,p=0．031。远期疗效中,肿瘤治愈组中ICAM-1mRNA和VEGFmRNA表达亦存在-定相关性,r=0．457,p=0．023;肿瘤复发组中IcAM-1mRNA和VEGFmRNA表达亦存在-定相关性,r=0．637,p=0．018。结论：ICAM-1高表达与宫颈鳞癌放疗抵抗性有关,并且与VEGF的表达在放射抵抗中具有-定的相关性。     

Caveolin-1 is down-regulated in alveolar rhabdomyosarcomas and negatively regulates tumor growth

Rhabdomyosarcoma is the most common soft tissue sarcoma of childhood and adolescence. Despite advances in therapy, patients with histological variant of rhabdomyosarcoma known as alveolar rhabdomyosarcoma (ARMS) have a 5-year survival of less than 30%. Caveolin-1 (CAV1), encoding the structural component of cellular caveolae, is a suggested tumor suppressor gene involved in cell signaling. In the present study we report that compared to other forms of rhabdomyosarcoma (RMS) CAV1 expression is either undetectable or very low in ARMS cell lines and tumor samples. DNA methylation analysis of the promoter region and azacytidine-induced re-expression suggest the involvement of epigenetic mechanisms in the silencing of CAV1. Reintroduction of CAV1 in three of these cell lines impairs their clonogenic capacity and promotes features of muscular differentiation. In vitro, CAV1-expressing cells show high expression of Caveolin-3 (CAV3), a muscular differentiation marker. Blockade of MAPK signaling is also observed. In vivo, CAV1-expressing xenografts show growth delay, features of muscular differentiation and increased cell death. In summary, our results suggest that CAV1 could function as a potent tumor suppressor in ARMS tumors. Inhibition of CAV1 function therefore, could contribute to aberrant cell proliferation, leading to ARMS development.     

Loss of heterozygosity on chromosome 6q27 and p53 mutations in epithelial ovarian cancer

Loss of heterozygosity (LOH) in chromosome region 6q27 and p53 mutations were studied to attempt to clarify the genetic etiology of ovarian cancer, with particular reference to clear cell adenocarcinoma (CCG), which has a poor prognosis. 6q27 LOH in 70 epithelial oyarian cancer patients was examined using four restriction fragment length polymorphism markers located at 6q27; p53 mutations in tumor DNA were detected using polymerase chain reaction singlestrand conformation polymorphism and sequencing. 6q27 LOH was confirmed in 26 of 48 informative cases (54.2%). No differences in the incidence of 6q27 LOH were seen by histologic type; 6q27 LOH was observed in 45% (5/11) of CCCs. p53 mutations were detected in 19 of the 48 tumors (39.6%), but in only one (9%) CCC. These results suggest that a putative tumor suppressor gene involved in the onset of epithelial ovarian cancer is located at 6q27. This gene is one of the keys to clarifying the genetic etiology of epithelial ovarian cancer and particularly CCC, given the low incidence of p53 mutations in this tumor type.     

上皮性卵巢肿瘤中3p14、3p25杂合性丢失

目的经对原发性上皮性卵巢癌的3号染色体短臂(3p14和3p25)杂合性丢失(LOH)分析研究,以探讨3pLOH与卵巢癌的发生发展之间的相关性.方法采用PCR法分别对50例原发性上皮性良恶性卵巢肿瘤(40例癌组织及10例非癌组织)的3p14及3P25处两个特定的位点D3S1228和D3S1038作杂合性丢失(LOH)检测.结果40例原发性上皮性卵巢癌中在3p处总LOH率为80%,其中3p14处发生LOH为24例(60.0%),3p25处为16例(40.0%),两处共同发生LOH的有8例(20.0%).FIGO卵巢癌的Ⅲa及Ⅲb期患者3pLOH率分别为93.8%及91.6%明显高于Ⅰ-Ⅱ期患者50.0%的3p丢失率(P＜0.05).10例良性卵巢肿瘤患者分别在3p14和3p25各处仅1例出现杂合性丢失,但未见有共丢失现象.40例不同病理类型的卵巢癌中以浆液性癌3p14和3p25LOH率为最高,但因病例数少而无统计学意义(P＞0.05).结论3p处可能存在一或多个与卵巢癌发生发展有关的侯选抑癌基因并与卵巢癌的恶性程度相关,有望作为遗传标志而应用于临床.