Human macrophages promote the motility and invasiveness of osteopontin-knockdown tumor cells

Increasing evidence indicates that macrophages in tumor stroma can significantly modify the malignant phenotypes of tumors. Osteopontin (OPN) is frequently overexpressed in cancers with high metastatic capacity and, thus, has been considered as a potential therapeutic target. To find out whether macrophages can affect the outcome of OPN-knockdown tumor cells, we used RNA interference (RNAi) to stably silence the OPN expression in the highly invasive human hepatoma cell line SK-Hep-1. Silencing of OPN markedly decreased the motility and invasiveness of the SK-Hep-1 cells. Further studies using this cell model revealed that coculture with human macrophages or macrophage-conditioned medium largely restored the migration and invasion potential of OPN-knockdown tumor cells. Moreover, such macrophage-promoted motility can be effectively blocked either by the addition of OPN-neutralizing antibody to the cocultured medium or by silencing OPN expression in macrophages. These results indicate that macrophage-derived OPN can compensate for the decrease of OPN and thereby restore the metastatic potential of OPN-knockdown tumor cells. Further characterization of the underlying mechanisms disclosed that macrophage-derived OPN exerted its function independently of the actin cytoskeleton rearrangement or the activation of matrix metalloproteinase and Rho families. Our results suggest that there are fine-tuned complex interactions between cancer cells and stroma cells, which may modify the outcome of cancer therapy, and therefore should be considered for the rational design of anticancer strategy.     

Paclitaxel inhibits ovarian tumor growth by inducing epithelial cancer cells to benign fibroblast-like cells

Paclitaxel is commonly used to treat multiple human malignancies, but its mechanism of action is still poorly defined. Human ovarian cancer SKOV3 cells (parental SKOV3) were treated with paclitaxel (1 μM) for 2 days, and the morphologic changes in the cells were monitored for more than 4 months. Parental SKOV3 underwent a markedly morphologic transition from the epithelial to fibroblast-like phenotype following treatment with paclitaxel; the resulting cells were designated as SKOV3-P. The SKOV3-P cells’ proliferative ability was assessed via a 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. The molecular characteristics of these cells were assessed via immunocytochemical staining and Western blot analysis. Their invasiveness and tumor formation ability was evaluated via wound-scratch and colony formation assays. The tumorigenicity of SKOV3-P cells was assessed in vivo after subcutaneous injection of tumor cells between injections of parental and paclitaxel-treated cells in nude mice. SKOV3-P cells have decreased the proliferation and invasion ability, decreased colony-forming ability when cultured in Matrigel and lost their tumor formation as compared with parental SKOV3 cells when injected in nude mice. SKOV3-P cells have decreased expression of E-cadherin, cytokeratin, Snail, PI3K, and P-Akt-Ser473, and increased expression of fibronectin, vimentin, Slug, P27, and PTEN. These results demonstrated that paclitaxel can inhibit tumor growth by inducing ovarian cancer epithelial cells toward a benign fibroblast-like phenotype through dysregulation of previously known pathways involved in the regulation of epithelial to mesenchymal transition (EMT), which may represent a novel mechanism for paclitaxel-induced tumor suppression.     

Cytokeratin 8 ectoplasmic domain binds urokinase-type plasminogen activator to breast tumor cells and modulates their adhesion, growth and invasiveness

Background  

Generation of plasmin is a characteristic of tumor cells, promoting the degradation of extracellular matrix, tumor progression and metastasis. The process is accelerated if plasminogen and plasminogen activator are bound to their cell surface receptors.     

Purified autoantibodies against glucose-regulated protein 78 (GRP78) promote apoptosis and decrease invasiveness of ovarian cancer cells

Circulating glucose-regulated protein 78 (GRP78) autoantibodies represent potential biomarker of epithelial ovarian cancer. However there is relatively limited identification of these antibodies in response to ovarian cancer. Here, we were interested in characterization of these antibodies and into their role in tumor development. We first purified GRP78 autoantibodies from sera of patients with ovarian cancer, and then tested them on SKOV-3 ovarian cancer cell line. A decrease of invasion and an increase of H(2)O(2)-induced apoptosis of SKOV-3 cells were observed, suggesting their protective role against ovarian cancer cells.     

Knockdown of polypyrimidine tract-binding protein suppresses ovarian tumor cell growth and invasiveness in vitro

Polypyrimidine tract-binding protein (PTB) is an RNA-binding protein with multiple functions in the regulation of RNA processing and IRES-mediated translation. We report here overexpression of PTB in a majority of epithelial ovarian tumors revealed by immunoblotting and tissue microarray (TMA) staining. By western blotting, we found that PTB was overexpressed in 17 out of 19 ovarian tumor specimens compared to their matched-normal tissues. By TMA staining, we found PTB expression in 38 out of 44 ovarian cancer cases but only in two out of nine normal adjacent tissues. PTB is also overexpressed in SV40 large T-antigen immortalized ovarian epithelial cells compared to normal human ovarian epithelial cells. Using doxycycline-inducible small interfering RNA technology, we found that knockdown of PTB expression in the ovarian tumor cell line A2780 substantially impaired tumor cell proliferation, anchorage-independent growth and in vitro invasiveness. These results suggest that overexpression of PTB is an important component of the multistep process of tumorigenesis, and might be required for the development and maintenance of epithelial ovarian tumors. Moreover, because of its novel role in tumor cell growth and invasiveness, shown here for the first time, PTB may be a novel therapeutic target in the treatment of ovarian cancer.     

红景天对乳腺癌抑制作用的研究

目的:通过体外和体内实验研究红景天对乳腺癌的增殖抑制作用和浸润转移相关因子表达的影响。方法:采用不同浓度红景天提取物处理乳腺癌MDA-MB-435细胞,制作细胞增殖曲线,观察药物对肿瘤细胞增殖活性的影响;应用流式细胞仪检测红景天提取物对乳腺癌细胞的细胞周期阻滞作用;选择裸鼠乳腺癌MDA-MB-435细胞移植瘤模型,以免疫组织化学染色方法研究红景天提取物对裸鼠移植瘤内MMP-2和组织蛋白酶D(CathD)表达的影响。结果:红景天提取物可显著抑制MDA-MB-435细胞的增殖活性,该抑制作用随着作用时间的延长、药物浓度的增高更为明显。红景天提取物具有阻滞MDA-MB-435细胞增殖周期的作用,主要阻滞于S期,该阻滞作用呈浓度依赖性。在红景天提取物治疗组裸鼠乳腺癌移植瘤中,CathD蛋白表达明显低于生理盐水对照组(免疫组化H-评分均数治疗组为72±50.70,而对照组为168±64.19,P=0.03);治疗组移植瘤MMP-2蛋白表达明显低于生理盐水对照组(免疫组化H-评分均数治疗组为90±29.15,而对照组为176±65.80,P=0.04)。结论:红景天可能通过抑制细胞增殖、浸润能力等多途径对乳腺癌细胞产生一定的抑制作用,其在乳腺癌的防治中具有一定的临床意义。     

Adipose-derived stem cells promote tumor initiation and accelerate tumor growth by interleukin-6 production

Adipose-derived stem cells (ADSCs) are multipotent cells that have attracted much recent attention. Here, we show that ADSCs enhance sphere formation and in vivo tumor initiation of breast and colon cancer cells. In co-culture, ADSCs induced several stem cell markers in cancer cells. ADSCs also accelerated tumor growth. Interaction of ADSCs and cancer cells stimulated secretion of interlukin-6 in ADSCs, which in turn acted in a paracrine manner on cancer cells to enhance their malignant properties. Interleukin-6 regulated stem cell-related genes and activated JAK2/STAT3 in cancer cells. We suggest that ADSCs may enhance tumor initiation and promotion.     

Effects of the proteinase inhibitor tetra-p-amidinophenoxyneopentane on in vitro adhesion and invasiveness of tumor cells

Some of the biological processes which enhance the metastatic capacity of tumor cells are associated with the activation of proteinases. In this paper we examined the effects of aromatic polyamidines on "in vitro" invasiveness of tumor cell lines. In addition to their antiproteinase activity, aromatic polyamidines exhibit antiproliferative activity toward tumor cell lines and inhibit expression of specific oncogenes. The results obtained show that the aromatic polyamidine TAPP-Br does not affect attachment, but inhibits the "in vitro" invasiveness of tumor cells cultured on a reconstituted basement membrane composed by laminin, type IV collagen, entactin, heparan sulphate proteoglycans. Our data suggest that this and related compounds should be further studied as experimental antitumor agents.     

Galectin-1 expression in cancer-associated stromal cells correlates tumor invasiveness and tumor progression in breast cancer

Understanding the molecular background of breast cancer biology is critical in developing new biomarkers for earlier diagnosis and more optimized treatment. We performed a proteomic analysis of human breast carcinoma tissues to investigate the tumor-specific protein expression in breast carcinoma. Using 2-dimensional electorphoresis (2-DE) and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS), we were able to identify a list of proteins which are upregulated in cancerous tissue. There was significant increase of galectin-1 expression in all cancerous tissues compared to noncancerous tissues, and its increased expression was further confirmed by western blot immunostaining. Subsequent immunohistochemical staining against galectin-1 in 105 breast cancer specimens showed significant correlation between galectin-1 expression in cancer-associated stromal cells and tumor invasiveness, T stage, TNM stage, and axillary lymph node metastasis. Galectin-1 expressionin cancer cells showed no correlation to above-mentioned pathologic variables. Hormonal receptor status and galectin-1 expression showed no correlation. This study demonstrates the upregulation of galectin-1 in breast carcinoma tissues and the clinical significance of galectin-1 in breast cancer patients. Our data supports the recently highlighted roles of galectin-1 in cancer-associated stroma and in tumor immune privilege.     

Differential regulation of growth and invasiveness of MCF-7 breast cancer cells by antiestrogens

Estrogen increases the ability of the estrogen-dependent MCF-7 human breast cancer cell line to both proliferate and invade through an artificial basement membrane. In studying the response of MCF-7 cells to various antiestrogens, we found that 4-hydroxytamoxifen and tamoxifen inhibited cell proliferation but increased their invasiveness. In contrast, the structurally unrelated benzothiophene antiestrogens, LY117018 and LY156758, were potent antiproliferative agents which did not stimulate invasiveness. The differential effects of these antiestrogenic agents on invasion correlated with changes in production of collagenase IV, while no significant change was seen in the chemotactic activity of the cells. Invasiveness was increased by 17 beta-estradiol or 4-hydroxytamoxifen after a few hours of treatment and was rapidly lost when 17 beta-estradiol was withdrawn. Stimulation of invasiveness with 17 beta-estradiol was blocked by the antiestrogen, LY117018. Cells from the MDA-MB-231 line which lacks estrogen receptors were not affected by estrogen or antiestrogen in terms of proliferation or invasion. These studies indicate that the invasiveness of MCF-7 cells is regulated by antiestrogens through the estrogen receptor and may be mediated by collagenase IV activity. Antiestrogens which reduce both the proliferation and invasiveness of these cells may be interesting new candidates for clinical application.     

Cathepsin S from both tumor and tumor‐associated cells promote cancer growth and neovascularization

Recent murine studies have demonstrated that tumor‐associated macrophages in the tumor microenvironment are a key source of the pro‐tumorigenic cysteine protease, cathepsin S. We now show in a syngeneic colorectal carcinoma murine model that both tumor and tumor‐associated cells contribute cathepsin S to promote neovascularization and tumor growth. Cathepsin S depleted and control colorectal MC38 tumor cell lines were propagated in both wild type C57Bl/6 and cathepsin S null mice to provide stratified depletion of the protease from either the tumor, tumor‐associated host cells, or both. Parallel analysis of these conditions showed that deletion of cathepsin S inhibited tumor growth and development, and revealed a clear contribution of both tumor and tumor‐associated cell derived cathepsin S. The most significant impact on tumor development was obtained when the protease was depleted from both sources. Further characterization revealed that the loss of cathepsin S led to impaired tumor vascularization, which was complemented by a reduction in proliferation and increased apoptosis, consistent with reduced tumor growth. Analysis of cell types showed that in addition to the tumor cells, tumor‐associated macrophages and endothelial cells can produce cathepsin S within the microenvironment. Taken together, these findings clearly highlight a manner by which tumor‐associated cells can positively contribute to developing tumors and highlight cathepsin S as a therapeutic target in cancer.     

Jagged-1 and Notch3 juxtacrine loop regulates ovarian tumor growth and adhesion

Notch3 gene amplification and pathway activation have been reported in ovarian serous carcinoma. However, the primary Notch3 ligand that initiates signal transduction in ovarian cancer remains unclear. In this report, we identify Jagged-1 as the highest expressed Notch ligand in ovarian tumor cells as well as in peritoneal mesothelial cells that are in direct contact with disseminated ovarian cancer cells. Cell-cell adhesion and cellular proliferation were reduced in Notch3-expressing ovarian cancer cells that were cocultured with Jagged-1 knockdown mesothelial and tumor feeder cells. Interaction of Notch3-expressing ovarian cancer cells with Jagged-1-expressing feeder cells activated the promoter activity of candidate Notch3 target genes, and this activity was attenuated by Notch3 siRNA. Constitutive expression of the Notch3 intracellular domain significantly suppressed the Jagged-1 shRNA-mediated growth inhibitory effect. In Notch3-expressing ovarian cancer cells, Jagged-1-stimulating peptides enhanced cellular proliferation, which was suppressed by gamma-secretase inhibitor and Notch3 siRNA. Taken together, our results show that Jagged-1 is the primary Notch3 ligand in ovarian carcinoma and Jagged-1/Notch3 interaction constitutes a juxtacrine loop promoting proliferation and dissemination of ovarian cancer cells within the intraperitoneal cavity.