Growth Of Human Ovarian Cancer Research Articles

The overexpressed regucalcin represses the growth via regulating diverse pathways linked to EGF signaling in human ovarian cancer SK-OV-3 cells: Involvement of extracellular regucalcin

AimsRegucalcin, which plays a multifunctional role in cell regulation, contributes as a suppressor in carcinogenesis. Survival of cancer patients is prolonged with high expression of regucalcin in tumor tissues. Ovarian cancer is the most lethal in gynecologic malignancies. This study elucidates the repressive role of regucalcin on the growth of human ovarian cancer SK-OV-3 cells that are resistant to cytotoxic cancer drugs. Materials and methodsSK-OV-3 wild type-cells and regucalcin-overexpressing cells (transfectants) were cultured in Dulbecco's Modification of Eagle's Medium containing 10 % fetal bovine serum. Key findingsColony formation and proliferation of SK-OV-3 cells were repressed by regucalcin overexpression. The suppressive effects of regucalcin on proliferation were independent of cell death. The proliferation of SK-OV-3 wild-type cells was repressed by various inhibitors, including cell cycle, signaling processes, and transcriptional activity. The effects of all inhibitors were not revealed in transfectants, suggesting the involvement of multiple signaling pathways in regucalcin effects. Of note, the overexpressed regucalcin declined the levels of Ras, Akt, mitogen-activating protein kinase, NF-κB p65, β-catenin, and STAT3, while it raised the levels of tumor suppressors p53 and Rb, and cell cycle inhibitor p21. Interestingly, the stimulatory effects of epidermal growth factor (EGF) on cell proliferation were blocked in regucalcin-overexpressing cells. Extracellular regucalcin repressed the proliferation independent of the death of SK-OV-3 cells and blocked EGF-enhanced cell proliferation. SignificancesThe overexpressed regucalcin may repress cell proliferation by targeting diverse signal pathways, including EGF signaling. This study offers a novel approach to the treatment of ovarian cancer with regucalcin.

Preparation of doxorubicin-loaded collagen-PAPBA nanoparticles and their anticancer efficacy in ovarian cancer.

The aims of this study were to prepare the collagen-poly (3-acrylamidophenylboronic acid) nanoparticles (collagen-PAPBA NPs) encapsulating doxorubicin (DOX) and research their anticancer efficacy in ovarian cancer. Collagen-PAPBA NPs were prepared, and their morphology and stability morphology were observed by transmission electron microscopy (TEM) and dynamic light scattering system (DLS). Preparation of doxorubicin-loaded Collagen-PAPBA NPs (DOX-loaded NPs) were then prepared, and the drug-loading content, encapsulation efficiency, and in vitro drug-release profiles were calculated. The morphology of DOX-loaded NPs was also observed by DLS, in vitro cytotoxicity to A2780 cells was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, in vitro antitumor activity on A2780 cells was observed by immunofluorescence, and in vivo antitumor activity was assessed using an experimental BALB/c mice tumor model. DOX-encapsulating collagen-PAPBA NPs were successfully prepared with mediation by biomolecule. The average hydrodynamic diameter of collagen-PAPBA NPs as measured by DLS was about 79 nm, with a homogeneous distribution of size. TEM revealed that nanoparticles were well-dispersed, spherical, and a roughly uniform 75 nm in size. Collagen-PAPBA NPs were quite stable in a wide range of pH and temperature conditions and associated with the concentration of glucose. DLS revealed that the average hydrodynamic diameter of DOX-loaded NPs was about 81.3 nm, with homogeneous distribution of size. TEM revealed that drug-loaded nanoparticles were spherical, well-dispersed, and gad a roughly uniform size of 79 nm. The proportion of DOX loaded into the nanoparticles was 10%, while the encapsulating efficiency was 97%. The result of the releasing test showed that the drug-loaded nanoparticles, as carriers for DOX, had a good sustained-release effect. The cell toxicity experiment showed that the blank NPs had no cytotoxicity to A2780 cells, and that the drug-loaded NPS had good a sustained-release function. They may thus have potential toxic-reducing side effects. Under the same doses, the drug-loaded NP had a superior inhibitory effect to free DOX on the growth of human ovarian cancer.

SiRNA-Mediated RRM2 Gene Silencing Combined with Cisplatin in the Treatment of Epithelial Ovarian Cancer In Vivo: An Experimental Study of Nude Mice.

Introduction: We aimed to explore small interfering (si)RNA silencing of ribonucleotide reductase M2 (RRM2) gene combined with cisplatin for the treatment of human ovarian cancer in nude mice models of subcutaneous transplantation of tumor cells.Methods: After conventional cultivation of human ovarian cancer cell line SKOV3 in vitro, SKOV3 cells were injected into the right back of nude mice by subcutaneous injection to establish the subcutaneous tumor models. Twenty-four tumor-burdened rats were randomly divided into four groups (n=6): siRNA group, siRNA in combination with cisplatin group, cisplatin group, and control group. Intraperitoneal injection of cisplatin and subcutaneous injection of siRNA were performed weekly. Tumor volume was measured, and tumor growth inhibition rate was calculated. RRM2 expression at the mRNA and protein levels was detected by reverse transcription-polymerase chain reaction and immunohistochemistry.Results: In the siRNA group, the tumor volume and tumor growth inhibition rate were 249.60±20.46 mm³ and 36.39%, respectively. The tumor growth inhibition rate and tumor volume were significantly different between the siRNA and control groups (p<0.05). In the cisplatin group, the tumor volume and tumor growth inhibition rate were 249.86±12.46 mm³ and 41.10%, respectively. The tumor growth inhibition rate and tumor volume were significantly different between the cisplatin and control groups (p<0.05). In the siRNA + cisplatin group, the tumor volume reduced to 180.84±16.25 mm³ and the tumor growth inhibition rate was increased to 64.33%, which were significantly different compared with the control group (p<0.01). Significant downregulation of RRM2 mRNA and protein expression in the tumor tissues was detected by reverse transcription polymerase chain reaction and immunohistochemistry assay (p<0.05).Discussion: siRNA alone or combined with cisplatin can effectively inhibit the growth of human ovarian cancer in nude mice models of subcutaneous transplantation of tumor cells. RRM2 gene silencing may be a potential treatment regimen for ovarian cancer in future.

CNDP2 Acts as an Activator for Human Ovarian Cancer Growth and Metastasis via the PI3K/AKT Pathway.

Introduction:The mechanism of tumorigenesis and metastasis of ovarian cancer has not yet been elucidated. This study aimed to investigate the role and molecular mechanism of cytosolic nonspecific dipeptidase 2 in tumorigenesis and metastasis.Methods:Cytosolic nonspecific dipeptidase 2 expression in human ovarian cancer tissues and cell lines was assessed with methyl thiazolyl tetrazolium (MTT), clone formation, and transwell assays performed to evaluate the ability of ovarian cancer cells to proliferate and migrate. Nude mice tumor formation experiments were also performed by subcutaneously injecting cells with stable cytosolic nonspecific dipeptidase 2 knockdown and control SKOV3 cells into BALB/c female nude mice to detect changes in PI3K/AKT pathway-related proteins by Western blotting.Results:Cytosolic nonspecific dipeptidase 2 was highly expressed in human ovarian cancer tissues, with its expression associated with pathological data, including ovarian cancer metastasis. A cytosolic nonspecific dipeptidase 2 stable knockdown or ectopic expression ovarian cancer cell model was established and demonstrated that cytosolic nonspecific dipeptidase 2 could promote the proliferation of ovarian cancer cells. Transwell cell migration and invasion assays confirmed that cytosolic nonspecific dipeptidase 2 enhanced cell metastasis in ovarian cancer. Furthermore, in vivo xenograft experiments demonstrated that cytosolic nonspecific dipeptidase 2 can promote the development and progression of ovarian cancer, increasing the expression of phosphorylated PI3K and AKT.Conclusions:Cytosolic nonspecific dipeptidase 2 promotes the occurrence and development of ovarian cancer through the PI3K/AKT signaling pathway.

Analysis of the inhibitory effects of miR-124 and miR-152 on human epithelial ovarian cancer xenografts in a nude mouse model.

This study investigated the inhibitory effects of miR-124 and miR-152 on the growth of human ovarian cancer (OC) SKOV3 cell line subcutaneous xenografts in nude mice. Twenty-eight healthy nude mice were selected and divided into the experimental group 1 (n=4), experimental group 2 (n=4), negative control group 1 (n=4), negative control group 2 (n=4), blank control group 1 (n=4), blank control group 2 (n=4) and observation group (n=4) according to the principle of similarity in body weight. The transfected SKOV3 cells were inoculated subcutaneously into the nape of the nude mice. After tumorigenesis, miR-124 mimics, miR-152 mimics, and their negative controls were transiently transfected into human OC SKOV3 cells via lipofection method. The expression levels of miR-124 and miR-152 were detected via reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and those of Ki-67 and caspase-3 were detected by western blotting. After transfection, the expression levels of miR-124 and miR-152 in the SKOV3 cells were significantly upregulated. The nude mice were sacrificed 36 days later, and tumor nodes of nude mice transfected with miR-124 and miR-152 grew slowly. Compared with that in the experimental groups, tumor size in the blank control and negative control groups was gradually increased with the increment of days (P<0.05). The volume of subcutaneous xenografts in nude mice of miR-124 and miR-152 experimental groups was obviously smaller than that in the blank control and negative control groups (P<0.05). Besides, the inhibition of tumor size in the observation group was more significant than that in the experimental groups (P<0.05). Thus, miR-124 and miR-152 inhibit the growth of human epithelial OC xenografts in nude mice, and they are expected to become new targets for gene-based therapy of OC.

Oldhamianoside inhibits the growth of ovarian cancer both in vitro and in vivo via adjusting inflammation and angiogenesis signals.

ObjectiveThe aim of this study was to determine the effects and possible mechanisms of oldhamianoside on the growth of human ovarian cancer both in vitro and in vivo.Materials and methodsCCK-8 assay was applied to estimate the effect of oldhamianoside on cell proliferation inhibition in vitro. Nude mice bearing human ovarian SKOV3 xenograft tumors were treated with oldhamianoside to investigate the effects of compound administration on tumor growth in vivo. To further investigate the mechanisms of inhibition effects of oldhamianoside on ovarian cancer growth in vivo, the levels of TNF-α, IL-6, and MCP-1 in plasma from the mice were measured by ELISA. Western blot was used to detect the expression of angiogenesis- and/or apoptosis-related proteins.ResultsWe found that oldhamianoside treatment inhibited SKOV3 proliferation and growth both in vitro and in vivo. Meanwhile, the levels of TNF-α, IL-6, and MCP-1 in plasma were markedly suppressed in oldhamianoside-treated mice. Additionally, oldhamianoside treatment inhibited the expression of VEGF and VEGFR2 and decreased the expression of caspase-3 and Bax/Bcl-2 ratio.ConclusionOur data indicate that oldhamianoside has an obvious inhibition effect on SKOV3 proliferation, and the mechanisms might be related to inhibition of cell growth, apoptosis induction, and adjusting the inflammatory response and angiogenesis signal.

Abstract 2913: Cancer growth under check: Anti-vacuolar ATPase "a2" antibody as novel therapy against ovarian cancer

Abstract Tumor growth and metastasis involves complex crosstalk among cancer cells and immune infiltrates. The balance of pro- and antitumor infiltrating immune cells can impact the outcomes of tumor growth. Vacuolar-ATPase (V-ATPase) proton pump promotes cancer growth and metastasis by regulating the pH-associated cancer cell-signaling and the tumor microenvironment. V-ATPase is overexpressed on cancer cell surface in a wide array of tumor types including ovarian cancer (OVCA) as well as on inflammatory cells recruited to the tumor microenvironment. This makes V-ATPase an important target for anticancer therapy. We have created a monoclonal antibody that specifically recognizes the plasma membrane isoform of V-ATPase (a2v) present distinctly on malignant cells and absent on normal cells. The purpose of this study is to investigate the in vivo effect of anti-VATPase-a2v antibody (Mab; 2C1) on the growth of human ovarian cancer (OVCA). The antitumor effects were evaluated in female athymic nude mice xenograft model. Mice were injected with human OVCA cell line (A2780;0.4X106 cells; s.c in upper flank) and the tumor growth was compared to mice that were simultaneously given a single dose of MAb (2C1; 300µg in PBS) or an isotype control antibody(mouse IgG1A). The 2C1 treatment resulted in delayed tumor growth (2.4 fold, p&amp;lt; 0.05), with no apparent in vivo toxicity. Histologic staining of 2C1-treated tumors revealed a loose tumor matrix with higher immune-infiltration compared to control. There was a marked decrease in cancer cell numbers in 2C1-treated tumor tissues compared to control using CA125 cancer-antigen staining. In in vitro assays, there was no measurable cytotoxic effect of 2C1 on OVCA cell proliferation, indicating the role of tumor microenvironment in delayed tumor growth. We further investigated the infiltrated immune population in 2C1-treated tumors. Interestingly, we observed an overall increased total leukocyte population (CD45; p=0.028) and a higher expression of iNOS (IHC-score 2C1 treated tumor= 11.3, control =7.6; p=0.04), suggesting an antitumor response. The total F4/80 macrophage population was higher in anti-VATPase-a2v antibody treated cells (absolute numbers in 2C1 treated tumor=245.2, control tumor=193.1; p=0.04). In 2C1 treated mice, tumor-associated macrophages displayed antitumor properties with correlated iNOS expression. Further, a marked increase in the number of Ly6G positive cells (neutrophil marker) was observed in 2C1 treated tumors, further suggesting an elevated antitumor response. Studies are under way to characterize other immune infiltrated populations that may contribute to antitumor response in 2C1 treated tumor tissues. In conclusion, the study demonstrates that the anti-VATPase "a2" antibody is an effective form of treatment in ovarian cancer. Citation Format: Arpita Kulshrestha, Gajendra K. Katara, Shayna Levine, Manoranjan Sahoo, Safaa A. Ibrahim, Alice Gilman-Sachs, Kenneth D. Beaman. Cancer growth under check: Anti-vacuolar ATPase "a2" antibody as novel therapy against ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2913.

Combined Gene Therapy Using AdsVEGFR2 and AdsTie2 With Chemotherapy Reduces the Growth of Human Ovarian Cancer and Formation of Ascites in Mice.

Ovarian cancer is highly dependent on tumor microvessels and angiogenesis regulated by vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) and angiopoietins (Ang) and their Tie receptors. We studied the efficacy of adenoviral (Ad) gene therapy with soluble VEGFR2 and Tie2 combined with paclitaxel and carboplatin for the treatment of ovarian cancer. An intraperitoneal human ovarian cancer xenograft model in nude mice (n = 44) was used in this study. Gene therapy was given intravenously when the presence of sizable tumors was confirmed in magnetic resonance imaging. The study groups were as follows: AdCMV as a control (group I), AdCMV with chemotherapy (group II), AdsVEGFR2 and AdsTie2 (group III), and AdsVEGFR2 and AdsTie2 with chemotherapy (group IV). Antitumor effectiveness was assessed by overall tumor growth, ascites, immunohistochemistry, microvessel density, and sequential magnetic resonance imaging analyses. AdsVEGFR2 and AdsTie2 gene therapy (group III) significantly reduced tumor weights as compared with group II (P = 0.007). Accumulation of ascites was significantly reduced when the mice were treated with AdsVEGFR2 and AdsTie2 gene therapy or with combined gene therapy and chemotherapy as compared with controls (P = 0.029 and P = 0.010, respectively). Vascular endothelial growth factor and Ang2 levels in ascites fluid were elevated after the gene therapy. Combined inhibition of VEGF/VEGFR2 and Ang/Tie2 pathways provided efficient therapy for ovarian cancer in mice. In addition, antiangiogenic gene therapy has potential as a treatment for the accumulation of ascites.

Abstract 4466: New biomarkers to optimize preclinical development of the PDI inhibitor XCE853

Abstract Protein disulfide isomerase (PDI) is a chaperone protein that regulates oxidative protein folding as well as cell viability. Increased PDI levels have been documented in a variety of human cancers, including ovarian, prostate, and lung cancers. Inhibition of PDI activity leads to apoptosis in cancer, suggesting that PDI is a promising druggable target. XCE853 was originally identified within a family of synthetic compounds displaying a preferential antiproliferative activity on drug-resistant human cancer cells (Gutmann et al, AACR annual meeting, 2013). XCE853 inhibits in vitro recombinant PDI activity and blocks in vitro proliferation of human tumor cells with IC50s in the nanomolar range through an irreversible cytolysis (Prevost et al, AACR annual meeting, 2014). By contrast, XCE853 has no or limited antiproliferative activity on several human non-tumoral immortalized cells (MCF10, RPE1 or RWPE1) even at high concentrations. The ex-vivo approach using fresh human hepatocellular carcinoma explants incubated 48h with XCE853 (100nM) has shown the decrease of the proliferation (KI-67 labeling) with an apoptosis induction (Caspase-3 labeling). In addition, XCE853 which displayed an excellent oral bioavailability in mouse blocks growth of human ovarian cancer using the in vivo OVCAR-3 xenograft model leading to complete tumor growth arrest even after the cessation of treatment. To optimize the preclinical and clinical development, we are now characterizing new biomarkers to 1) stratify the tumors which are sensitive and non-sensitive to XCE853 and to 2) identify the optimal dose regimen. Altogether, these data support further efforts on XCE853 preclinical studies to prepare the Phase-I clinical study entry. Citation Format: Gregoire Pierre Prevost, Marine Garrido, Shili Xu, Celine Lefebvre, Anne Chauchereau, Denis Carniato, Jean François Briand, Mathieu Gutmann, Maria Serova, Annemilai Tijeras-Raballand, Armand De Grammont, Eric Raymond, Christian Gespach, Michèle Sabbah, Nouri Neamati, Marc-Henry Pitty, Paul Foster. New biomarkers to optimize preclinical development of the PDI inhibitor XCE853. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4466. doi:10.1158/1538-7445.AM2015-4466

Potential new chemotherapy strategy for human ovarian carcinoma with a novel KSP inhibitor

Among synthetic kinesin spindle protein (KSP) inhibitor compounds, KPYB10602, a six-member lactam-fused carbazole derivative was the most potent in vitro against cell growth of human ovarian cancer, A2780. KPYB10602 caused dose-dependent suppression of tumor growth in vivo. Mitotic arrest due to KPYB10602 was confirmed in vitro, and was characterized by inhibition of securin degradation. Apoptosis after mitotic arrest was associated with an increase in the ratio of pro-apoptotic Bax to anti-apoptotic Bcl-2. Increase of reactive oxygen species (ROS) and caspase pathway were also involved. Furthermore, KPYB10602 caused little neurotoxicity in vivo. Therefore, KPYB10602 could be a promising candidate as an anti-tumor agent with reduced adverse events for treating human ovarian cancer.

Paris saponin II inhibits human ovarian cancer cell-induced angiogenesis by modulating NF-κB signaling.

The clinical applications of Rhizomaparidis in traditional Chinese medicine are well known. However, the therapeutic potential of Rhizomaparidis and its active component such as Paris saponinI (polyphyllin D) and Paris saponinII (PSII) (formosaninC) in cancer treatments have not yet been fully explored. Recent studies have demonstrated that PSII and chemoagents exhibit comparable inhibitory affects against human ovarian cancer cell growth. Since NF-κB, a ubiquitous transcription factor that plays an important role in cancer biology, is often associated with gynecological cancers, in the present study, we evaluated the possibility that PSII modulates NF-κB activity and VEGF-mediated angiogenesis and elucidated the molecular mechanisms underlying such effects. We assessed the effects of PSII on NF-κB activity in SKOV3 tumor cells and on tumor cell induced-angiogenesis using standardized angiogenesis invitro, exvivo and invivo assays, western blot analysis and kinase assay. We also assessed the effect of the super-engineered repressor of IĸBα and its effect, in combination with PSII treatment on tumor growth and angiogenesis in xenograft athymic mouse models of ovarian cancer (SKOV3 and SKOV3/mutant IĸBα cells) using color Doppler ultrasound and traditional immunohistochemistry. We showed that PSII suppressed NF-κB activation as a result of the reduction in IKKβ kinase activity on its substrate IκBα and the expression of IKKβ. Compromising NF-κB activation reduced the expression of NF-κB-downstream targets such as VEGF, Bcl-2 and Bcl-xL. Such inhibitory effects at molecular levels appear to compromise tumor growth and angiogenesis. Most importantly, the combination of PSII treatment and constitutive repression of IĸBα activity exhibited marked inhibitory effects against human ovarian cancer cell growth in a xenograft mouse model of ovarian cancer. For the first time, we provide evidence showing that PSII potently inhibits angiogenesis and the growth of human ovarian cancer by suppressing NF-κB signaling.

Abstract 1812: XCE853: A novel PDI inhibitor that inhibits proliferation of human tumor cells in vitro, ex-vivo and in vivo

Abstract XCE853 was originally identified within a family of synthetic compounds displaying a preferential antiproliferative activity on drug resistant human cancer cells (Gutmann et al, AACR annual meeting, 2013). XCE853 induced an irreversible cytolysis of human tumor cells after a short in vitro exposure independently of efflux pumps leading to a tumor cell death by autophagy and particles release (vesicles or protein aggregates). The interest for XCE853 has been reinforced by three in vitro observations: 1) XCE853 inhibits the recombinant human enzyme PDI recognized as one novel target linked to drug resistance 2) XCE853 has a unique antiproliferative profile compared to the NCI database compounds when using the NCI Compare assay and 3) XCE853 has no activity on human normal prostate cells even at high concentrations whereas XCE shows an IC50 in the nanomolar range on human tumoral cells. In addition, the ex-vivo approach using fresh human tumor explants cultivated in 3 dimensions with low concentrations of XCE853 has shown a strong decrease of the proliferation (KI-67 labeling) with an apoptosis induction (Caspase-3 labeling) of Head &amp; Neck cancers following 24h of exposure. Finally, XCE853 which displayed an excellent oral bioavailability in mouse is able to block growth of human ovarian cancer using in vivo xenograft model leading to a complete tumor growth arrest even after the stop of the treatment. Altogether, these data support further efforts on this drug candidate to initiate the preclinical studies and to define the most relevant human tumor types. Citation Format: Gregoire P. Prevost, Shili Xu, Marine Garrido, Maria Serova, Olivier DE Vewer, Christian Gespach, Jean-François Briand, Annemilaï Tijeras-Raballand, Mathieu Gutmann, An Hendrix, Michele Sabbah, Anne Chachereau, Armand de Gramont, Nouri Neamati, Denis Carniato, Marc-Henry Pitty, Paul Foster. XCE853: A novel PDI inhibitor that inhibits proliferation of human tumor cells in vitro, ex-vivo and in vivo. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1812. doi:10.1158/1538-7445.AM2014-1812

Journal of Cellular Biochemistry: Volume 115, Number 11, November, 2014

Journal of Cellular BiochemistryVolume 115, Issue 11 p. C1-C1 CoverFree Access Journal of Cellular Biochemistry: Volume 115, Number 11, November, 2014 First published: 10 September 2014 https://doi.org/10.1002/jcb.24971AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Abstract Cover: Efficacy of S. typhimurium A1-R on orthotopic growth of human ovarian cancer in nude mice. See article by Matsumoto et al. on pages 1996–2003. Volume115, Issue11November 2014Pages C1-C1 RelatedInformation

Efficient inhibition of intraperitoneal human ovarian cancer growth by short hairpin RNA targeting CD44.

CD44 is one member of a big glycoprotein family involved in adhesion of cells or cells and extracellular matrix (ECM). The heavily glycosylated CD44 has been proved to be a major receptor of hyaluronan and a marker of stem cells in ovarian cancer. Here, using short hairpin (shRNA) against CD44, we demonstrate that knockdown CD44 could inhibit cancer growth efficiently compared with controls. Plasmid targeting CD44 gene (pshCD44) or non-relative control sequences (pshHK) was constructed and delivered to ovarian cancer by biodegradable poly D, L-Lactide-co-glycolide acid nanoparticles (PLGANPs). Nude mice were utilized in an intraperitoneal model of ovarian carcinomatosis to assess antitumor efficacy in vivo. Antitumor efficacy was estimated by changes in tumor weights, proliferation (Ki-67), apoptosis (TUNEL) and angiogenesis (CD31 staining and alginate-encapsulated tumor beads assay) in tumor cells. As results, pshCD44 or pshHK could be effectively transfected into SKOV-3 cells by PLGANPs. Tumor weight in pshCD44/PLGANPs group was suppressed by 45% and 50% compared with those in pshHK/PLGANPs and untreated group, respectively (Ps < 0.001). Inhibition of cell proliferation, induction of apoptosis and reduction of angiogenesis in tumor cells of pshCD44/PLGANPs group also show significant difference compared with those in control groups (Ps < 0.05), respectively. These results indicate that pshCD44 delivered by PLGANPs might be a potential approach in ovarian cancer therapy, and point towards a mechanism involving the inhibition of angiogenesis, cellular proliferation and the induction of apoptosis.

PGPIPN, a Therapeutic Hexapeptide, Suppressed Human Ovarian Cancer Growth by Targeting BCL2

Bioactive peptides, either derived from nature resources or synthesized by rational design, have been demonstrated potential for therapeutic agents against numerous human diseases, including cancer. However, the mechanism of therapeutic peptides against cancer has not been well elucidated. Here we show that PGPIPN, a hexapeptide derived from bovine β-casein, inhibited the proliferation of human ovarian cancer cells line SKOV3 as well as the primary ovarian cancer cells in vitro. Consistently, PGPIPIN also decreased tumor growth rate in xenograft ovarian cancer model mice in a dose-dependent manner. Further study demonstrated that the anti-tumor effect of PGPIPN is partially through promoting cell apoptosis by inhibiting BCL2 pathway. Thus, our study suggests that PGPIPN is a potential therapeutic agent for the treatment of ovarian cancer or other types of cancer.

Inhibitory Effect of Ethyl Acetate Extract of the Shaggy Inc Cap Medicinal Mushroom, Coprinus comatus (Higher Basidiomycetes) Fruit Bodies on Cell Growth of Human Ovarian Cancer

In a previous study, we screened organic extracts of different mushroom mycelia and picked ethyl acetate extract of Coprinus comatus as one of the most active extracts against human ovarian cancer cells. In the current study, we extracted a dry powder of C. comatus fruit bodies using ethyl acetate and examined its effect on the viability of three cell lines originated from human ovarian cancer (ES-2, SKOV-3, and SW-626). This extract was active against all tested cell lines, in a dose-dependent manner (concentrations 50-200 µg/mL, P<0.01). In an attempt to segregate the active fraction, we subjected the extract to chromatography on a silica gel column. The effect of six different fractions and of the crude extract on the viability of ES-2 cells was examined after exposure time of 24 h. Fraction F (last eluted) was significantly more effective than crude extract in the reduction of cell viability (P<0.01). Fraction F was also significantly more active than crude extract in the reduction of viability of SKOV-3 cells. We next identified some of the compounds of fraction F (mainly fatty acids) by gas chromatography-mass spectrometry. In summary, ethyl acetate extract of C. comatus reduced viability of three lines of human ovarian cancer. Fractionation of this extract by a silica gel column enabled the selection of a fraction significantly more active than the original extract.

Expression of CD40 and growth-inhibitory activity of CD40 agonist in ovarian carcinoma cells.

The CD40 receptor is a member of the tumour necrosis factor receptor family and is widely expressed on various cell types. The antitumour activity of CD40 agonist antibody has been observed in B-cell-derived malignancies, but its activity on ovarian cancer remains unclear. However, in this paper, we first confirmed that the anti-CD40 agonist antibody could inhibit the growth of ovarian cancer cells and induce apoptosis. This study investigated the expression of CD40 by ovarian carcinoma tissues and cell lines, at the same time, we evaluated the effect of a recombinant soluble human CD40L (rshCD40L) and an anti-CD40 agonist antibody on cell growth and apoptosis. Flow cytometry and immunohistochemistry assay demonstrated that CD40 was expressed on ovarian carcinoma cell lines and primary ovarian carcinoma cells derived from ascites, as well as on ovarian carcinoma tissues. The growth inhibition of rshCD40L and the anti-CD40 agonist antibody on ovarian carcinoma cells was examined by MTT assay, and the proportion of apoptotic tumour cells was analysed by flow cytometry and Hoechst staining. Our study showed that CD40 was expressed on all ovarian carcinoma cell lines and was examined in 86.2% (162/188) of ovarian cancer tissue samples, but not in normal ovarian tissues (n=20). Treatment with rshCD40L or anti-CD40 agonist antibody significantly inhibited ovarian carcinoma cell growth and induced apoptosis. Theses results suggest that CD40 is expressed on ovarian carcinoma cells, moreover, that rshCD40L and anti-CD40 agonist antibody have therapeutic potential to inhibit human ovarian cancer growth.

Growth Hormone-Releasing Hormone Antagonists Inhibit Growth of Human Ovarian Cancer

Epithelial ovarian carcinoma is the leading cause of cancer-related deaths among women with gynecologic malignancies. Antagonists of the growth hormone-releasing hormone (GHRH) have been shown to inhibit growth of various cancers through endocrine, autocrine, and paracrine mechanisms. In this study, we have investigated the effects of GHRH antagonists (GHRHa) in ES-2 human clear cell ovarian cancer and in UCI-107 human serous ovarian cancer in vitro and in vivo. We evaluated the expression of mRNA for GHRH receptor, the binding to GHRH receptors, in specimens of ES-2 ovarian cancer. We evaluated also the in vitro effects of GHRHa on ES-2 cells and the in vivo effect of 2 different GHRHa on ES-2 and UCI-107 tumors. Nude mice bearing xenografts on ES-2 and UCI-107 ovarian cancer were treated with JMR-132 and MZ-J-7-118, respectively. Tumor growth was compared to control. ES-2 cells expressed mRNA for the functional splice variant SV1 of the GHRH receptor. JMR-132 inhibited cell proliferation in vitro by 42% and 18% at 10 and 1 μM concentration, respectively. Specific high affinity receptors for GHRH were detected in ES-2 cancer samples. In vivo daily subcutaneous injections of GHRHa significantly reduced tumor growth compared to a control group in both animal models. Our results indicate that GHRHa such as JMR-132 and MZ-J-7-118 can inhibit the growth of human ovarian cancer. The efficacy of GHRHa in ovarian cancer should be assessed in clinical trials.

Combination of ciglitazone, a peroxisome proliferator-activated receptor gamma ligand, and cisplatin enhances the inhibition of growth of human ovarian cancers

We have recently reported that peroxisome proliferator-activated receptor gamma (PPARγ) ligands produce antitumor effects against human ovarian cancer in conjunction with reduction in angiogenesis and induction of apoptosis via regulating prostaglandin (PG) E(2) level. In this study, we investigated the effects of combination of ciglitazone, a PPARγ ligand, and cisplatin, a cytotoxic anti-cancer drug, on growth of ovarian cancer. Tumor growth and survival were examined in female nu/nu mice xenografted with subcutaneous OVCAR-3 tumors or with intraperitoneal DISS tumors and treated with cisplatin alone (5mg/kg intraperitoneally once on day 1), ciglitazone alone (15mg/kg intraperitoneally once a week), or the combination. Ciglitazone alone, cisplatin alone, or their combination significantly suppressed the growth of OVCAR-3 tumors xenotransplated subcutaneously and prolonged the survival of mice with malignant ascites derived from DISS cells as compared with the control. Furthermore, the combination produced a significantly greater antitumor effect than cisplatin or ciglitazone alone and also significantly prolonged the survival time as compared with cisplatin or ciglitazone alone. The combination significantly decreased PGE(2) concentration in serum as well as in ascites, reduced vascular endothelial growth factor as well as microvessel density, and induced apoptosis in solid OVCAR-3 tumor as compared with cisplatin or ciglitazone alone. The combination remarkably decreased the expression of cyclooxygenase-2 (COX-2), microsomal PG E synthase (mPGES), and PG receptor 3 (EP3) in tumors. In vitro experiment showed that ciglitazone enhances the cytotoxicity of cisplatin against ovarian cancer cells. In conclusion, the combination inhibited the growth of ovarian cancer in conjunction with reduction in angiogenesis and induction of apoptosis resulting from suppression of PGE(2) activation through decreasing the expression of COX-2, mPGES, and EP3. The inhibitory effect of this combination treatment on growth of ovarian cancer suggests a potential to lead a novel therapeutic strategy against ovarian cancer.

Knockdown of stat3 expression by RNAi inhibits in vitro growth of human ovarian cancer

BackgroundThe aim of the study was to investigate the suppressive effects of pSilencer2.1-U6-siRNA-stat3 recombinant plasmids on the growth of ovarian cancer in vitro.Material and methods.Three pairs of DNA template (stat3-1, stat3-2, stat3-3) specific for different target sites on stat3 mRNA were synthesized to reconstruct pSilencer2.1-U6-siRNA-stat3s, which were transfected into SKOV3 cells. The expressions of STAT3, BcL-2, cyclin D1 and C-myc in these cells were detected by Western blot and Northern blot. The cell cycle and the growth were determined by flow cytometry (FCM) and MTT assay, respectively. Cell apoptosis was determined by TUNEL staining.ResultsOf the three siRNAs, only siRNA targeting stat3-3 markedly suppressed the protein expression of stat3 in SKOV3 cells; MTT assay and FCM showed that transfection of stat3-3 siRNA could significantly suppress the growth of SKOV3 cells and arrest the cell cycle in vitro. TUNEL staining also showed massive apoptosis in SKOV3 cells transfected with stat3-3 siRNA.ConclusionspSilencer2.1-U6-siRNA-stat3-3 can significantly inhibit the STAT3 expression in human ovarian cancer cells resulting in the inhibition of the cancer growth and the increase of apoptosis of cancer cells.