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As a kind of powerful anti-counterfeiting device, diffractive optically variable image (DOVI) has been developed and widely used in information security field. However, the identification of DOVI today by bare eyes is not reliable. In this paper we investigate the recognition of DOVI with machine learning method, and five kinds of algorithms, namely quadratic discriminate analysis (QDA), linear discriminate analysis (LDA), regularized discriminate analysis (RDA), leave-one-out covariance matrix estimate (LOOC), and Kullback-Leibler information measure based method (KLIM) are applied to the recognition of DOVI. Considering both time cost and correct classification rate, KLIM classifier exceeds others.

The aim of this study is to investigate the plasticity of human epithelial ovarian cancer cell SKOV3ip and formation of vasculogenic mimicry (VM) in vivo. SKOV3ip was transfected with lentiviral vector carrying green fluorescence protein (GFP). Female nude mice were implanted intraperitoneally with GFP-labled SKOV3ip. When the transplanted tumor reached a volume of approximately 1 cm3, paraffin-embedded, formaldehyde-fixed tissue was prepared and stained with hematoxylin and eosin (H & E). Tumor tissues were also studied by electron microscopy and fluorescence microscopy. The results of H & E staining, electron microscopy, and fluorescence microscopy indicated SKOV3ip formed patterned networks with erythrocytes in them, in the absence of vascular epithelial cells, which was a sign that SKOV3ip engaged in VM in vivo. Expression of vascular epithelium marker CD31 was investigated by immunohistochemical staining, immunofluorescence assay, semiquantitative reverse transcriptase–polymerase chain reaction (RT-PCR), and flow cytometric analysis (FACS). Factor VIII and vascular endothelial growth factor (VEGF) were also analyzed by FACS. Weak and focal CD31 immunohistochemical staining was found along the channels of tumor cells. Immunofluorescence assay and RT-PCR demonstrated that CD31 was expressed in primary-cultured SKOV3ip. CD31 and Factor VIII, but not VEGF were detected in primary-cultured SKOV3ip by FACS. The present study has shown that human ovarian cancer cell line SKOV3ip may be able to express some specific markers of vascular epithelial cells and has plasticity to form VM in vivo. In the following study, we indicated that hypoxia-inducible factor (HIF)-1α inhibitor, rapamycin, could possibly prevent VM and phenotype transformation of SKOV3ip, reflected by down-regulating expression of CD31 and Factor VIII. HIF-1α protein expression correlated with CD31 and Factor VIII protein expression in SKOV3ip. These results indicated that VM might be associated with HIF-1α.

Progesterone plays an important role in the regulation of normal endometrium function by binding to progesterone receptor (PR). In endometrial cancer, however, PR is always down-regulated. Previous reports showed that methylation in the promoter region of the PR gene may be responsible for PRB isoform repression. However, the CpG islands in the exon region of the PR gene are much richer and longer than in the promoter region. We hypothesize that methylation in the exon region may also take part in the down-regulation of the PR gene. The methylation status of the first exon of the PR gene in endometrial cell cultures was investigated. Aberrant methylation patterns were observed in the first exon of PR gene, and the methylation density is correlated with the differentiation of different types of endometrial cancer cells. DNA methyltransferase (DNMT) and histone deacetylase inhibitor 5-aza-2'-deoxycytidine (ADC), as well as trichostatin A (TSA), which reverses PR gene expression, were also studied. A combination of ADC and TSA resulted in synergistic effects in inducing PR expression, down-regulation of DNMT1 and DNMT3A, and could also have antigrowth effect on endometrial cancer cells by inducing apoptosis.

BACKGROUND Endometrial cancer is a common gynecologic malignancy among women. The molecular mechanisms involved in the progression of endometrial cancer are unclear, which has hampered the development of an effective treatment. Survivin, a newly identified member of the inhibitor of apoptosis (IAP) family, regulates 2 critical processes in neoplastic transformation: cell proliferation and apoptosis. METHODS Survivin mRNA and protein expression levels were analyzed in human normal cycling endometrium, atypical endometrium, and endometrial adenocarcinoma by immunohistochemical, reverse-transcriptase polymerase chain reaction (RT-PCR), and Western blot analyses. To study the biological function of survivin in endometrial cancer, RNA interference was applied to knock down survivin expression in the Ishikawa endometrial cancer cell line by recombinant plasmids producing survivin small hairpin RNA. Furthermore, the signal pathway that regulates survivin expression was investigated. RESULTS Higher levels of survivin mRNA and protein expression were observed in endometrial adenocarcinomas than in atypical or normal endometrium. Immunohistochemical staining revealed that 83.3% (50 of 60) of endometrial adenocarcinoma samples, 55.0% (11 of 20) of atypical endometrium samples, and 25.0% (5 of 20) of normal endometrium samples were positive for survivin protein. Inhibition of survivin by RNA interference reduced cell proliferation and induced apoptosis in Ishikawa cells by down-regulating cyclin D1 and phosphorylated RB and activating caspase-3 and caspase-8. The authors also found that the MAPK pathway was a signal transduction pathway upstream of survivin. Epidermal growth factor (EGF) and transforming growth factor-α (TGF-α) up-regulated survivin protein expression by activating the MAPK pathway in endometrial cancer cells. CONCLUSIONS Survivin is an attractive target for endometrial cancer treatment. Growth factors could regulate survivin expression by activating the MAPK pathway. Cancer 2006. © 2006 American Cancer Society.

The aims of this study were to investigate the hypoxia-inducible factor 1alpha (HIF-1alpha) protein inhibition and tumor growth by a molecular target of rapamycin inhibitor, rapamycin, in xenogeneic transplant model of ovarian cancer and to study the correlation of apoptosis with HIF-1alpha and vascular endothelial growth factor (VEGF) expression. Four groups of female nude mice were inoculated subcutaneous with SKOV-3 cells and treated with vehicle, rapamycin, paclitaxel, or rapamycin plus paclitaxel. The expressions of HIF-1alpha and VEGF and microvessel density (MVD) were assessed by immunohistochemistry. While messenger RNA (mRNA) expression of Glut1, bcl-2, and VEGF was studied by reverse transcription-polymerase chain reaction, and apoptosis of tumor cells was determined by terminal deoxynucleotidyl biotin-dUTP nick end labeling (TUNEL). The HIF-1alpha was expressed in epithelial ovarian cancer. There was a significant correlation between HIF-1alpha protein expression and VEGF or MVD. Tumor burden treated with rapamycin alone, rapamycin plus paclitaxel, and paclitaxel alone was reduced (47.91%, 51.03%, and 31.75%, respectively) compared with controls. The expression of HIF-1alpha was inhibited, and apoptotic index of tumor cell increased in rapamycin and rapamycin plus paclitaxel group. HIF-1alpha may upregulate VEGF expression both in mRNA and protein level. There is a positive correlation between HIF-1alpha and MVD. Rapamycin inhibits expression of HIF-1alpha and suppresses ovarian tumor growth. Our data suggested that a combination of HIF-1alpha inhibitor and chemotherapy could provide an effective approach for inhibiting tumor growth in ovarian cancer.

To identify an effective auxiliary therapy for epithelial ovarian cancer.Progesterone acetate given at 250 mg intramuscularly twice a week for 1 month followed by increased administration to 500 mg intramuscularly every two weeks for 3 years was used in combination with platinum based chemotherapy to treat patients with epithelial ovarian cancer as a first-line therapy. Prognoses of the patients receiving progesterone combined with chemotherapy (progesterone group) and those receiving chemotherapy only (control group) were compared.Three-year recurrence and survival conditions of the progesterone and control groups were as follows. Stage Ia: no patient relapsed or died in either group. Stage Ib-Ic: three-year recurrence rates were 14.2% and 37.5%, respectively (P = 0.2845); three-year survival rates were 92.3% and 87.5% (P = 0.7221). Stage II: 1 patient relapsed and died among the 3 patients in the progesterone group; among the 4 patients in the control group, 1 patient relapsed, none died. Stage III: three-year recurrence rates were 30.8% and 64.3%, respectively (P = 0.1170); three-year survival rates were 85.7% and 42.9%, respectively (P = 0.005). Stage IV: 4 patients relapsed and 1 patient died among the 7 patients in the progesterone group; both the patients in the control group relapsed and died.The results indicated that progesterone combined with platinum based chemotherapy as a first-line therapy may improve the prognosis of advanced epithelial ovarian cancer, but would not change the prognosis of early stage epithelial ovarian cancer.

To investigate the effects of sulindac metabolites on proliferation and apoptosis in the human umbilical vein endothelial cell line ECV304 in vitro.The proliferation profile of ECV304 was determined by methyl thiazolyl tetrazolium (MTT) method. Cell cycle distribution, apoptosis and the ultrastructure of ECV304 were detected by flow cytometry (FCM) and electron microscopy, respectively.MTT assay showed that the sulfide inhibited the proliferation of ECV304 and its effect was dose-dependent; the IC(50) was 200 micromol/L. FCM showed that the sulfide changed cell cycle distribution. The cell cycle distribution was as follows: G(1) phase (control group 77.74% +/- 1.58%; sulfone group 75.63% +/- 2.12%; sulfide group 46.12% +/- 1.60%); S phase (control group 13.64% +/- 1.22%; sulfone group 16.40 +/- 2.30%; sulfide group 27.26% +/- 2.08%); G(2)-M phase (control group 8.61% +/- 0.67%; sulfone group 7.98% +/- 0.49%; sulfide group 26.62% +/- 3.54%). The apoptosis rates in the control group, sulfone group and sulfide group were 6.08% +/- 3.39%, 4.81% +/- 2.14% and 51.90% +/- 5.67%, respectively. Sulfide reduced the proportion of G(1) phase, increased the proportion of S phase, G(2)-M phase and the apoptosis rate significantly (P0.01, vs control). In the sulfide-treated cells, there were nuclear fragmentation and chromosomal condensation, shrinkage of the cell and loss of contact with neighboring cells. Apoptotic bodies were observed. Sulfone showed no effect on cell proliferation, cell cycle distribution or cell morphology.Sulfide can significantly reduce the proliferation of ECV304, change the cell cycle distribution and arrest cells in G(2)-M phase where apoptosis may be induced. Sulfone has no such effects on this cell line.

To investigate the effects of sulindac metabolites on the proliferation and apoptosis of human umbilical vein endothelial cell line ECV304 in vitro.The proliferation of ECV304 was determined by methyl thiazolyl tetrazolium (MTT) method. The cell cycle, apoptosis and the ultrastructure of ECV304 were detected by flow cytometry (FCM) and electron microscopy respectively.MTT assay showed that sulfide inhibited the proliferation of ECV304, and the effects was dose-dependent, the 50% inhibiting concentration (IC(50)) was 200 micromol/L. FCM showed that sulfide changed cell cycle distribution, the cell cycle were: Go-G(1) phase [control group (77.7 +/- 1.6)%, sulfone group (75.6 +/- 2.1)%, sulfide group (46.1 +/- 1.6)%]. S phase [control group (13.6 +/- 1.2)%, sulfone group (16.4 +/- 2.3)%, sulfide group (27.3 +/- 2.1)%], G(2)-M phase [control group (8.6 +/- 0.7)%, sulfone group (8.0 +/- 0.5)%, sulfide group (26.6 +/- 3.5)%]. The apoptosis rates in control group, sulfone group and sulfide group were (6.1 +/- 3.4)%, (4.8 +/- 2.1)% and (51.9 +/- 5.7)%, respectively. Compared with the control group, sulfide can reduce the proportion of G(1) phase, increase the proportion of S phase and G(2)-M phase significantly (P0.01). Also sulfide induce cell apoptosis significantly (P0.01). In the sulfide-treated cells, chromatin concentration, cytomembrane shrinkage, neighboring cell detachment, and apaptolic body were observed. Sulfone has no effect on cell proliferation, cell cycle and cell morphology.Sulfide can reduce the viability of ECV304, change the cell cycle distribution, arrest the cell cycle in G(2)-M phase and induce apoptosis. Sulfone didnet.

Epithelial ovarian cancer (EOC) remains the most lethel gynecological malignancy. In the absence of an effective screening strategy, two thirds of patients are diagnosed in advanced clinical stage. Despite chemotherapy with cisplatin and paclitxel, primary or acquired chemoresistance faciltates survival of cancer cells, resulting in disease recurrence and death. Our previous studies indicate that ovarian cancer patients whose tumors overexpress transient receptor potential channel C3 (TRPC3) protein tend to relapse early and have shorter overall survival. In vitro studies indicated that follicle-stimulating hormone (FSH) induced resistance to cisplatin-induced apoptosis could be partially reversed by blocking expression of TRPC3 in EOC cell lines. To enhance sensitivity to platinum compounds, we have investigated the relationship between resistance to cisplatin and TRPC3. We have now found that expression of TRPC3 in recurrent cancer following chemotherapy is higher than in specimens from primary cytoreductive surgery. The expression of TRPC3 in the cisplatin-resistant EOC line A2780cp is higher than in the sensitive parental cell line A2780. An in vitro study showed that transient treatment of the EOC cell line HEY with cisplatin upregulated TRPC3 mRNA and membrane localized protein. Induced cisplatin-resistant cell line HEYcp augmented the expression of TRPC3 and enhanced invasive capacity in cell migration assays. Knockdown of TRPC3 expression in EOC cell lines with specific siRNA or miRNA decreased cell migratory ability and enhanced cisplatin-induced apoptosis when compared to EOC cells treated with scrambled control RNA. Our data indicate that TRPC3 may play an important role in cisplatin-resistance of EOC and could serve as a therapeutic target to resensitize cancer cells to cisplatin in culture and in xenografts. Citation Format: Xiang Tao, Hongyan Jin, Zhenbo Zhang, Robert C. Bast, Youji Feng, Yinhua Yu. Downregulation of TRPC3 enhances sensitivity to cisplatin and inhibits motility in epithelial ovarian cancer. [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 3762. doi:10.1158/1538-7445.AM2014-3762

Ovarian cancer is the most lethal malignancy of the female reproductive system with an overall mortality rate of greater than 70%. Our group has demonstrated that Follicle stimulating hormone (FSH) stimulates the proliferation and invasion of ovarian cancer cells, inhibits apoptosis, and facilitates neovascularization by increasing expression of vascular endothelial growth factor (VEGF). OCT4, a stem cell marker, is overexpressed in several types of human cancer and can induce resistance to chemotherapy and inhibition of apoptosis. We have found that FSH stimulates the expression of OCT4 mRNA and protein in a time- and dose-dependent manner. Subcellular analysis by immunofluorescent staining confirmed that FSH stimulation increased OCT4 protein expression in both the cytoplasm and nucleus of ovarian cancer cells. Re-expression of OCT4 in ovarian cancer cells reversed the inhibition of apoptosis induced by FSH, whereas siRNA-mediated knockdown of OCT4 resulted in a significantly rate of apoptosis. Depletion of OCT4 resulted in reduction of p-AKT and survivin, and also blocked FSH-induced upregulation of p-AKT and survivin. Moreover, FSH stimulation induced the expansion of CD44+CD117+ cells with a stem cell-like phenotype. Interestingly, re-expression of OCT4 enhanced the expression of Notch, Sox2, and Nanog, molecules that play critical roles in cancer stem cell proliferation and differentiation. FSH upregulated the expression of Notch, Sox2 and Nanog and these effects were abolished by knocking down OCT4, suggesting that several cancer stem cell pathways are involved in FSH regulation. We have also examined OCT4 expression in surgical specimens of ovarian cancer. Immunohistostaining revealed that OCT4 expression was increased in ovarian carcinoma compared with benign cystadenomas and borderline tumors, and OCT4 expression was significantly correlated with histological grade. Staining for OCT4 was increased in serous cystadenocarcinoma, when compared with clear cell carcinoma. In summary, the OCT4 cancer stem cell signaling pathway may mediate FSH-induced inhibition of apoptosis and could provide a target for treatment of ovarian cancer. Citation Format: Zhenbo Zhang, Yaping Zhu, Xianming Xu, Xiaowei Xi, Robert C. Bast, Yinhua Yu, Youji Feng. Follicle stimulating hormone (FSH) inhibits apoptosis in ovarian cancer cell through regulating OCT4 stem cell signal pathway. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4766. doi:10.1158/1538-7445.AM2013-4766

Objective To study the relationship between cell proliferation and apoptosis in cervical carcinoma and its clinical significance.

At present, epithelial ovarian cancer is the leading killer among all gynecological malignancies. Despite much research, development of this neoplasm is still not fully understood. The five-year survival rate for this disease still remains about 30%. Recent studies suggest that follicle-stimulating hormone (FSH) may play a major role in ovarian epithelial carcinogenesis. Our group has demonstrated that FSH stimulates the proliferation and invasion of ovarian cancer cells, inhibits apoptosis, and facilitates neovascularization by increasing vascular endothelial growth factor (VEGF) expression. To understand the mechanisms by which FSH stimulates ovarian cancer growth, we have performed gene expression arrays and confirmed changes in protein expression by Western blots. Interestingly, we found that FSH stimulation enhances expression of transient receptor potential channel C3 (TRPC3) at both mRNA and protein levels. Silencing TRPC3 expression by siRNA inhibited the ability of FSH to stimulate proliferation of ovarian cancer cell lines and to block apoptosis. TRPC channels are nonselective cation channels with permeability to Ca2+. Increases in cytosolic free Ca2+ can promote different physiological changes including cell growth, differentiation and death. TRPC3 is one of 7 TRPC family members. Our previous work has shown that TRPC3 contributes to the progression of human ovarian cancer. TRPC3 protein levels in human ovarian cancer specimens were greatly increased relative to those in normal ovarian specimens. Downregulating TRPC3 expression led to reduced proliferation, suppression of epidermal growth factor-induced Ca2+ influx, and decreased growth of xenografts. In this study, we further explored the interaction of FSH-TRPC3. FSH stimulation was associated with upregulation of TRPC3 levels and translocation of TRPC3 from the cytoplasm to cell membrane, facilitating the influx of Ca2+ after treatment with a TRPC3 specific agonist. Knocking down TRPC3 abrogated Akt/PKB phosphorylation by FSH stimulation, leading to decreased expression of down-stream effectors including survivin, HIF1α and VEGF. Our findings indicate that TRPC3 plays a significant role in the tumor-stimulating activity of FSH and could be a potential therapeutic target of ovarian cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4540. doi:10.1158/1538-7445.AM2011-4540

Follicle-stimulating hormone (FSH) has been demonstrated to increase the risk of ovarian malignancy, but the mechanisms remains unclear. Our previous results showed that FSH increases the expression of VEGF through activation of survivin, which is mediated by the activation of the PI3K/AKT signaling pathway. We reported, herein, that FSH promoted ovarian cancer cells proliferation and prevented apoptosis in vitro and in vivo. FSH up-regulated the expression of survivin and down-regulated the expression of programmed cell death 6 (PDCD6) and death receptor 5 (DR5), two molecules required for apoptosis induction. Further, we have defined that the decreased expression of DR5, but not PDCD6, was mediated by survivin. We have established animal xenografts tissue arrays, the tumors from the groups with FSH treatment had higher expression of survivin and lower expression of DR5 comparing with the tumors from untreated groups, further confirming the interaction of FSH-survivin-DR5. In addition, we have demonstrated that FSH does not only inhibit apoptosis, but also inhibit autophagy. FSH inhibits autophagy by activating mTOR pathway in ovarian cancer cells under different stress conditions, including serum starvation. By Reverse Phase Protein Arrays (RPPA) analysis and western blot confirmation, we found that FSH stimulation increases the level of Rab25 in multiple ovarian cancer cells in a dose dependent manner. Rab25, a small GTPase, is found to be associated with the progression of ovarian tumor. Rab25, belongs to Rab11 supfamily, was shown to involve in endosomal recycling. To study the role of FSH-Rab25 interaction in regulating autophagy, we knocked down Rab25 expression in ES-2, Hey and SKOv3 ovarian cancer cell lines by siRNA, and examined the effects of FSH on autophagy. Rab25 knockdown increased the starvation induced autophagy through the inhibition of pAkt, while FSH administration could partly reverse the effects of Rab25 knockdown, due to the increase of Rab25 expression stimulated by FSH. Our findings indicated that Rab25 is one of the molecules regulated by FSH in ovarian cancer cells that has a novel role as a regulator in autophagy. Taken together, we have demonstrated that FSH promotes ovarian cancer development through regulating expression of survivin, Rab25, PDCD6 and DR5, which in turn regulates cell proliferation, apoptosis as well as autophagy. Further understanding the molecular mechanisms of FSH in the process of ovarian carcinogenesis and development will ultimately help developing an effective cancer prevention method, and find a novel targeted therapy for ovarian cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3666.

Background p53 is the most commonly mutated tumor suppressor gene in human cancers. In addition to the loss of tumor suppression function and exertion of dominant-negative effects over the remaining wild-type protein, several p53 mutants can gain novel oncogenic functions (gain-of-function, GOF) that actively regulate cancer development and progression. In human endometrial cancer, p53 mutation is more often associated with aggressive nonendometrioid cancer. However, it was unknown if p53 mutants contributed to endometrial cancer progression through the GOF properties. Methods To clarify the relationship between expression of p53 GOF mutation (p53-R175H) and invasive potential of human endometrial cancer KLE cells, we tested the consequences of up-regulation and down-regulation of p53-R175H in KLE cells by inducing p53-R175H expression vector or suppressing the p53 gene with short hairpin RNA. Results We found that forced over-expression of p53-R175H significantly promoted cell migration and invasion, and induced activation of the epidermal growth factor receptor (EGFR)/phosphatidylinositol 3-kinase (PI3K)/AKT pathway. Conversely, suppression of p53-R175H with short hairpin RNA significantly inhibited cell migration and invasion, and resulted in attenuation of EGFR/PI3K/AKT pathway. Conclusion These findings show for the first time that elevated expression of p53-R175H mutant may exert gain-of-function activity to activate the EGFR/PI3K/AKT pathway and thus may contribute to the invasive phenotype in endometrial cancer.

Background IQGAP1 is a scaffolding protein and overexpressed in many human tumors, including ovarian cancer. However, the contribution of IQGAP1 to invasive properties of ovarian cancer cells remains unknown. Here, we investigated the effect of IQGAP1-specific short hairpin RNA (shRNA) expressing plasmids on metastatic potential of ovarian cancer HO-8910PM cells. Methods We used RT-PCR and Western blot analysis to characterize expression of IQGAP1 in three human ovarian cancer-derived cell lines SK-OV-3, HO-8910 and HO-8910PM. We then determined whether expression of endogenous IQGAP1 correlated with invasive and migratory ability by using an in vitro Matrigel assay and cell migration assay. We further knocked down IQGAP1 using shRNA expressing plasmids controlled by U1 promoter in HO-8910PM cells and examined the proliferation activity, invasive and migration potential of IQGAP1 shRNA transfectants using MTT assay, in vitro Matrigel-coated invasion assay and migration assay. Results IQGAP1 expression level seemed to be closely associated with the enhanced invasion and migration in ovarian cancer cell lines. Levels of both IQGAP1 mRNA and protein were significantly reduced in HO-8910PM cells transfected with plasmid-based IQGAP1-specific shRNAs. RNAi-mediated knockdown of IQGAP1 expression in HO-8910PM cells resulted in a significant decrease in cell invasion and migration. Conclusion Our findings support the hypothesis that IQGAP1 promotes tumor progression and identify IQGAP1 as a potential therapeutic strategy for ovarian cancer and some other tumors with over-expression of the IQGAP1 gene.