Heparanase expression and localization in different types of human lung cancer

Background

Heparanase is the only known mammalian glycosidase capable of cleaving heparan sulfate chains. The expression of this enzyme has been associated with tumor development because of its ability to degrade extracellular matrix and promote cell invasion.

Methods

We analyzed heparanase expression in lung cancer samples to understand lung tumor progression and malignancy. Of the samples from 37 patients, there were 14 adenocarcinomas, 13 squamous cell carcinomas, 5 large cell carcinomas, and 5 small cell carcinomas. Immunohistochemistry was performed to ascertain the expression and localization of heparanase.

Results

All of the tumor types expressed heparanase, which was predominantly localized within the cytoplasm and nucleus. Significant enzyme expression was also observed in cells within the tumor microenvironment, such as fibroblasts, epithelial cells, and inflammatory cells. Adenocarcinomas exhibited the strongest heparanase staining intensity and the most widespread heparanase distribution. Squamous cell carcinomas, large cell carcinomas, and small cell carcinomas had a similar subcellular distribution of heparanase to adenocarcinomas but the distribution was less widespread. Heparanase expression tended to correlate with tumor node metastasis (TNM) staging in non-small cell lung carcinoma.

Conclusion

In this study, we showed that heparanase was localized to the cytoplasm and nucleus of tumor cells and to cells within the microenvironment in different types of lung cancer. This enzyme exhibited a differential distribution based on the type of lung tumor.General significanceElucidating the heparanase expression patterns in different types of lung cancer increased our understanding of the crucial role of heparanase in lung cancer biology. This article is part of a Special Issue entitled Matrix-mediated cell behaviour and properties.     

肺腺癌发生阻塞性肺炎的临床分析

目的研究阻塞性肺炎与肺腺癌发病的关系以及对预后的影响。方法对2003年12月至2013年1月于大连医科大学附属二院确诊的436例肺腺癌患者病历进行回顾,根据患者临床特征进行分组,并进行统计学分析。结果晚期肺腺癌患者更易出现阻塞性肺炎（χ2=5．662,P=0．017）;中心型肺腺癌较周围型更易出现阻塞性肺炎（χ2=4．432,P=0．035）;在肺腺癌中,性别、年龄、吸烟史以及体重下降与否与阻塞性肺炎的发生经比较差异无统计学意义（P〉0．05）。结论阻塞性肺炎的发生与晚期肺腺癌以及中心型肺腺癌有关,重视筛查对肺腺癌的诊断和预后有重要意义。     

Outcome of a multimodal therapy of a recurrent adenocarcinoma arising from Caesarean section scar endometriosis—A case report

Background

Endometriosis occurring in surgical scars is a well-described entity. Malignant transformation of endometriosis is a rare event, with most cases belonging to adenocarcinoma. The initial surgical treatment is a method of choice. Due to lack of therapeutic recommendations, adjuvant therapy and recurrence management are a great challenge for oncologists.

Aim

The aim of this paper was to present a long-term survival as the outcome of multimodal therapy in the patient with recurrent adenocarcinoma arising from Caesarean section scar endometriosis.

Case

We present the case of a woman with recurrent adenocarcinoma arising from Caesarean section scar endometriosis. The disease was first diagnosed in September 1997 at age 43. The patient underwent abdominal hysterectomy with tumour excision. Due to a local recurrence after 4 years, tumour excision with abdominal wall repair using a plastic mesh, regional lymphadenectomy, bilateral salpingo-ovariectomy and adjuvant radiotherapy for the pelvic region with local boost were performed; in addition hormontherapy with medroxyprogesterone was started. Because of a recurrent pelvic tumour, chemotherapy, further local palliative radiotherapy and brachytherapy were administered. Subsequently distant metastases in bilateral axillary lymph nodes were diagnosed and palliative radiotherapy was performed. The patient died of morbus neoplasmaticus generalisatus in September 2008. The follow-up period had been 132 months.

Conclusion

This paper is, to our knowledge, the only report in literature that presents a long-term survival as the outcome of multimodal therapy in the patient with this rare diagnosis. Further reports of new cases can help establish optimal treatment guidelines.     

The MUC4 membrane-bound mucin regulates esophageal cancer cell proliferation and migration properties: Implication for S100A4 protein

MUC4 is a membrane-bound mucin known to participate in tumor progression. It has been shown that MUC4 pattern of expression is modified during esophageal carcinogenesis, with a progressive increase from metaplastic lesions to adenocarcinoma. The principal cause of development of esophageal adenocarcinoma is the gastro-esophageal reflux, and MUC4 was previously shown to be upregulated by several bile acids present in reflux. In this report, our aim was thus to determine whether MUC4 plays a role in biological properties of human esophageal cancer cells. For that stable MUC4-deficient cancer cell lines (shMUC4 cells) were established using a shRNA approach. In vitro (proliferation, migration and invasion) and in vivo (tumor growth following subcutaneous xenografts in SCID mice) biological properties of shMUC4 cells were analyzed. Our results show that shMUC4 cells were less proliferative, had decreased migration properties and did not express S100A4 protein when compared with MUC4 expressing cells. Absence of MUC4 did not impair shMUC4 invasiveness. Subcutaneous xenografts showed a significant decrease in tumor size when cells did not express MUC4. Altogether, these data indicate that MUC4 plays a key role in proliferative and migrating properties of esophageal cancer cells as well as is a tumor growth promoter. MUC4 mucin appears thus as a good therapeutic target to slow-down esophageal tumor progression.     

Egr-1基因与肿瘤腺癌放射敏感性关系的研究

目的：研究Egr-1基因在2种腺癌细胞A549和Hela放射前后的表达变化及对放射敏感性的影响。方法：培养肺腺癌A549细胞和宫颈腺癌Hela细胞,分别提取4Gyx射线照射前及照射后不同时间点的细胞的总RNA行荧光定量PCR（FQ-PCR）检测Egr-1表达水平;收获4GyX射线照射前及照射后不同时间点的细胞处理行流式细胞术检测其凋亡;对照射不同剂量的细胞继续培养10-14天,进行克隆形成计数,计算克隆形成率及存活分数。结果：FQ-PCR结果显示,放射后Egr．1基因表达水平在2种细胞中均明显升高且于放射后1h达到峰值,A549细胞的峰值明显高于Hela细胞;流式细胞术检测结果显示,A549细胞凋亡明显高于Hela细胞;克隆形成实验结果显示,A549细胞存活分数明显低于Hela细胞。结论：Egr-1基因在不同腺癌细胞表达水平不同并影响其放射敏感性。     

Cytotoxic studies of substituted titanocene and ansa-titanocene anticancer drugs

A variety of substituted titanocene and ansa-titanocene complexes have been synthesized and characterized using traditional methods. The cytotoxic activity of the different titanocene complexes was tested against tumour cell lines human adenocarcinoma HeLa, human myelogenous leukemia K562, human malignant melanoma Fem-x and normal immunocompetent cells, peripheral blood mononuclear cells PBMC. Alkenyl substitution, either on the cyclopentadienyl ring or on the silicon-atom ansa-bridge of the titanocene compounds [Ti{Me₂Si(η⁵-C₅Me₄)(η⁵-C₅H₃{CMe₂CH₂CH₂CHCH₂})}Cl₂] (8), [Ti{Me(CH₂CH)Si(η⁵-C₅Me₄)(η⁵-C₅H₄)}Cl₂] (9) and [Ti(η⁵-C₅H₄{CMe₂CH₂CH₂CHCH₂})₂Cl₂] (12) showed higher cytotoxic activities (IC₅₀ values from 24 ± 3 to 151 ± 10 μM) relative to complexes bearing an additional alkenyl-substituted silyl substituent on the silicon bridge [Ti{Me{(CH₂CH)Me₂SiCH₂CH₂}Si(η⁵-C₅Me₄)(η⁵-C₅H₄)}Cl₂] (10) and [Ti{Me{(CH₂CH)₃SiCH₂CH₂}Si(η⁵-C₅Me₄)(η⁵-C₅H₄)}Cl₂] (11) which causes a dramatic decrease of the cytotoxicity (IC₅₀ values from 155 ± 9 to >200 μM). In addition, the synthesis of the analogous niobocene complex [Nb(η⁵-C₅H₄{CMe₂CH₂CH₂CH=CH₂})₂Cl₂] (13), is described. Structural studies based on DFT calculations of the most active complexes 8, 9 and 12 and the X-ray crystal structure of 13 are reported.     

Study of the cytotoxic activity of di and triphenyltin(IV) carboxylate complexes

The reaction of 3-methoxyphenylacetic acid (3-MPAH), 4-methoxyphenylacetic acid (4-MPAH), 2,5-dimethyl-3-furoic acid (DMFUH) or 1,4-benzodioxane-6-carboxylic acid (BZDOH) with triphenyltin(IV) chloride (1:1) or diphenyltin(IV) dichloride (2:1) in the presence of triethylamine yielded the compounds [SnPh₃(3-MPA)] (1), [SnPh₃(4-MPA)] (2), [SnPh₃(DMFU)] (3), [SnPh₃(BZDO)] (4), [SnPh₂(3-MPA)₂] (5), [SnPh₂(4-MPA)₂] (6), [SnPh₂(DMFU)₂] (7) and [SnPh₂(BZDO)₂] (8), respectively. The tetranuclear complex [{Me₂(DMFU)SnOSn(DMFU)Me₂}₂] (9) was prepared by the reaction of dimethyltin(IV) oxide and 2,5-dimethyl-3-furoic acid (DMFUH). The molecular structures of 3, 4 and 9, were determined by X-ray diffraction studies. The cytotoxic activity of the carboxylic acids (3-MPAH, 4-MPAH, BZDOH and DMFUH) and di (5-8) and triphenyltin(IV) complexes (2-4) was tested against tumor cell lines human adenocarcinoma HeLa, human myelogenous leukemia K562, human malignant melanoma Fem-x and normal immunocompetent cells, peripheral blood mononuclear cells PBMC. Triphenyltin(IV) complexes show higher activities than the diphenyltin(IV) derivatives. The most active compound is [SnPh₃(DMFU)] (3) with IC₅₀ value of 0.15 ± 0.01, 0.051 ± 0.004, 0.074 ± 0.004, 0.20 ± 0.01, 0.15 ± 0.02 on HeLa, K562, Fem-x, rested and stimulated PBMC, respectively, while the most selective are [SnPh₂(3-MPA)₂] (5), [SnPh₂(DMFU)₂] (7) and [SnPh₂(BZDO)₂] (8). Compounds 3, 5, 7 and 8 present higher activities than cisplatin in all the tested cells and relative high selectivity especially on K562 cells.     

CD15 mRNA和nm23H1 mRNA表达与结直肠癌转移和预后的关系

为了探讨评估结直肠癌预后指标。应用催化信号放大-原位杂交技术。研究90例结直肠癌组织中CD15和nm23H1的mRNA表达,并结合随访资料分析,结果表明,在结直肠癌中CD15和nm23H1的mRNA阳性表达分别为84.4%和66.7%。CD15mRNA高表达及nm23H1mRNA低表达与结直肠癌浸润程度,淋巴结转移和肝脏转移状况密切相关,提示CD15和nm23H1的mRNA表达均可作为预测结直肠癌侵袭转移和客观评估患者预后的生物学指标。     

Mitochondrial alterations induced by serum amine oxidase and spermine on human multidrug resistant tumor cells

Summary. Multidrug resistance (MDR) has been studied extensively because it is one of major problems in cancer chemotherapy. The MDR phenotype is often due to overexpression of P-glycoprotein (P-gp), that acting as an energy-dependent drug efflux pump exports various anticancer drugs out of cells. The major goal of our investigation is to establish whether bovine serum amine oxidase (BSAO), which generates the products H₂O₂ and aldehyde(s), from the polyamine spermine, is able to overcome MDR of human cancer cells. The cytotoxicity of the products was evaluated in both drug-sensitive (LoVo WT) and drug-resistant (LoVo DX) colon adenocarcinoma cells. A clonogenic cell survival assay demonstrated that LoVo DX cells were more sensitive than LoVo WT cells. Exogenous catalase protected cells against cytotoxicity mainly due to the formation of H₂O₂. However, spermine-derived aldehyde(s) still induced some cytotoxicity. The cytotoxic effect was totally inhibited in the presence of both enzymes, catalase and NAD-dependent aldehyde dehydrogenase (ALDH). Transmission electron microscopy investigations showed that BSAO and spermine induced evident mitochondria alterations, more pronounced in MDR than in LoVo WT cells. The mitochondrial activity was checked by flow cytometry studies, labelling cells with the probe JC1, that displayed a basal hyperpolarized status of the mitochondria in multidrug-resistant cells. After treatment with amine oxidase in the presence of polyamine-spermine, the cells showed a marked increase in mitochondrial membrane depolarization higher in LoVo DX than in LoVo WT cells. Our findings suggest that toxic oxidation products formed from spermine and BSAO could be a powerful tool in the development of new anticancer treatments, mainly against MDR tumor cells.     

Seroreactivity to 19.5-kDa antigen in combination with absence of seroreactivity to 35-kDa antigen is associated with an increased risk of gastric adenocarcinoma

Background. Only a minority of those infected with Helicobacter pylori will develop gastric cancer. Stratification of H. pylori strains based on carcinogenic potential will provide a basis for selective surveillance and eradication therapy. We studied the anti‐H. pylori antibody profile in Asian patients with gastric adenocarcinoma to identify any H. pylori antigen that may be associated with an increased or decreased risk of gastric carcinoma. Patients and Methods. A case‐control study comparing the seroprevalence of antibodies with various H. pylori antigens in Singaporeans with gastric adenocarcinoma and the normal Singaporean population was carried out using both conventional immunoglobulin (Ig) G enzyme‐linked immunosorbent assay (ELISA) and Western blot immunoassay. Results. The seroprevalence among 44 gastric adenocarcinoma cases (70.5% males, mean age 66.7 ± 13.5 years) and 261 controls (49.8% males, mean age 61.5 ± 4.1 years) was 90.9% vs. 50.2% by IgG ELISA. In the H. pylori‐positive male population, those suffering from gastric adenocarcinoma had significantly lower seroreactivity to the 35‐kDa antigen compared with asymptomatic controls (p = .0198, OR = 3.79, 95% CI 1.24–11.61). Seropositivity to the 19.5 kDa antigen was also found to be associated with the presence of gastric adenocarcinoma in Singaporean males (p = .022, OR = 4.17, 95% CI 1.22–14.28). A ‘high‐risk’ phenotype consisting of absence of a band at 35‐kDa in combination with the presence of a band at 19.5‐kDa was significantly associated with the presence of gastric adenocarcinoma (p = .002, OR = 3.7, 95% CI 1.6–8.6). Conclusions. Stratification of H. pylori strains based on their potential for carcinogenesis, such as those strains that are seropositive for the 19.5 kDa antigen and seronegative for the 35‐kDa antigen, may provide a basis for selective eradication of H. pylori infection and future vaccine development.