Migration Of SKOV3 Cells Research Articles

Discovery of HB-EGF binding peptides and their functional characterization in ovarian cancer cell lines

Ovarian cancer is one of the most frequent causes of cancer death among all gynecologic cancers. Though standard therapy often results in temporary clinical remission, most patients suffer from recurrence and metastasis of ovarian cancer, which highlights the need for developing new therapeutic agents targeting specific molecules. Previous studies have demonstrated that the native ligand of epidermal growth factor receptor (EGFR) and ErbB4, heparin-binding EGF-like growth factor (HB-EGF), plays a critical role in the progression of ovarian cancer and is associated with prognosis of ovarian cancer. In the current study, we tried to develop a peptide-based treatment for ovarian cancer by targeting HB-EGF. After the functions of HB-EGF in promoting migration and invasion of SKOV3 and HO-8910 cells were confirmed, phage display was used to discover peptides binding to HB-EGF. Two peptides, no. 7 and no. 29 were found mildly binding to HB-EGF. Then the effects of these peptides on HB-EGF functions were examined and both peptides no. 7 and no. 29 were found indeed inhibiting the functions of HB-EGF in promoting migration and invasion of SKOV3 and HO-8910 cells in vitro. Further mechanism investigation showed that peptides no. 7 and no. 29 inhibited HB-EGF-promoted cell migration and invasion through attenuating activation of the EGFR signaling pathway manifested by decreased p-Erk1/2 and Snail levels. More importantly, peptides no. 7 and no. 29 showed strong activities in inhibiting migration of SKOV3 cells in vivo. These results provide a proof of concept method for developing novel peptide drugs to combat ovarian cancer through interfering with HB-EGF mediated signaling pathways.

TM4SF1 is a potential target for anti-invasion and metastasis in ovarian cancer

BackgroundPatients with ovarian cancer commonly have a poor prognosis, owing to its invasiveness and distant metastasis. Studies have found TM4SF1 participates in regulating tumor cell invasion and migration. Therefore, it is expected to become a target for anti-invasion and metastasis in ovarian cancer.MethodsThe expression of TM4SF1 in normal ovarian epithelial tissues, benign ovarian tumor tissues, primary foci of epithelial ovarian cancer and the matched lymph mode metastatic foci was detected using immunohistochemistry to analyze its association with prognosis. The expression of TM4SF1 in HO8910PM, SKOV3 was inhibited using RNAi, and the growth, proliferation, migration, invasion abilities of HO8910PM and SKOV3 cells and the growth of xenograft tumors in nude mice were examined.Results(1) The positive expression rate of TM4SF1 protein in epithelial ovarian cancer tissues (90.90%) was higher than that in benign ovarian tumor tissues (65.22%) and normal ovarian epithelial tissues (31.25%), and both differences were significant (P < 0.05). The expression of TM4SF1 protein was positive in all metastatic lymph node foci and matched primary foci (100%). (2) The level of TM4SF1 protein expression was positively correlated with the International Federation of Gynecology and Obstetrics (FIGO) stage and histological grade. However, The positive TM4SF1 protein expression was not an independent factor of prognosis (P > 0.05). (3) Silencing TM4SF1 expression did not affect growth, proliferation, or cell cycle distribution but inhibited the migration and invasion abilities of HO8910PM and SKOV3 cells. Silencing TM4SF1 expression inhibited the growth of xenograft tumors in nude mice.ConclusionTM4SF1 is a potential target for anti-invasion and metastasis in ovarian cancer.

MiR-539-3p promotes the progression of epithelial ovarian cancer by targeting SPARCL1.

The aim of this study was to investigate the effect of microRNA-539-3p (miR-539-3p) on the development of epithelial ovarian cancer (EOC), and to explore the possible underlying mechanism. A total of 40 paired EOC tissues and adjacent normal ovarian tissues were surgically resected in Hanchuan People's Hospital. Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of miR-539-3p in EOC tissues and cell lines. Targeted regulatory mechanism of miR-539-3p on SPARC-like protein 1 (SPARCL1) was identified by luciferase reporter and Western blot assays. Furthermore, the effects of miR-539-3p/SPARCL1 axis on the malignant behaviors of EOC cells, including proliferation, invasion and migration abilities, were confirmed by cell counting kit-8 (CCK-8), transwell and scratch wound assays. QRT-PCR showed that the expression of miR-539-3p was significantly up-regulated in EOC tissues and cell lines. SPARCL1 was a direct target of miR-539-3p in EOC cells. Overexpression of miR-539-3p significantly promoted the proliferation, migration and invasion of SKOV3 cells. Furthermore, co-transfection of miR-539-3p inhibitor and si-SPARCL1 could remarkably restore the migration and invasion abilities of SKOV3 cells. MiR-539-3p acted as an oncogene in EOC by targeting SPARCL1. MiR-539-3p/SPARCL1 axis, as a target for the treatment of EOC, might become a feasible and new method of tumor treatment.

TRPM7 promotes the epithelial\u2013mesenchymal transition in ovarian cancer through the calcium-related PI3K / AKT oncogenic signaling

BackgroundThe epithelial-mesenchymal transition (EMT) is crucial for metastasis and positively regulated by calcium-related signaling. The melastatin-related transient receptor potential 7 (TRPM7) regulates a non-selective cation channel and promotes cancer metastasis. However, the mechanisms underlying the action of TRPM7 in ovarian cancer are unclear.MethodsThe expression of TRPM7 and EMT markers (Vimentin, N-cadherin, Twist and E-cadherin) in ovarian cancer samples was detected. TRPM7was knockdown by shRNA in Ovarian cancer cell lines to examine calcium [Ca2+]i, EMT markers and PI3K/AKT markers. Various cellular assays, such as invasion and migration, were performed in vitro, and further confirmed in vivo.ResultsTRPM7 expression is negatively correlated with E-cadherin, but positively with N-cadherin, Vimentin and Twist expression in ovarian cancer samples. TRPM7 depletion inhibited the migration and invasion in SKOV3 and OVCAR3 cells. In addition, TRPM7 silencing decreased the lung metastasis of SKOV3 tumors and prolonged the survival of tumor-bearing mice. Similar to that of TRPM7 silencing, treatment with MK886, a potent 5-lipoxygenase inhibitor to reduce TRPM7 expression, and/or BAPTA-AM, an intracellular calcium chelator, significantly mitigated the Epidermal growth factor (EGF) or Insulin-like growth factors (IGF)-stimulated migration, invasion, and the EMT in ovarian cancer cells by decreasing the levels of intracellular calcium [Ca2+]i. Furthermore, treatment with LY2904002, a PI3K inhibitor, also inhibited the migration, invasion, and treatment with both LY2904002 and BAPTA-AM further enhanced their inhibition in ovarian cancer cells. Moreover, treatment with BAPTA-AM mitigated the IGF-stimulated migration, invasion, particularly in TRPM7-silenced ovarian cancer cells. Finally, TRPM7 silencing attenuated the PI3K/AKT activation, which was enhanced by BAPTA-AM, MK886 or LY2904002 treatment in ovarian cancer cells.ConclusionsTRPM7 silencing inhibited the EMT and metastasis of ovarian cancer by attenuating the calcium-related PI3k/AKT activation. Our findings suggest that TRPM7 may be a therapeutic target for intervention of ovarian cancer.

Peptide P7 inhibits the bFGF-stimulated proliferation and invasion of SKOV3 cells.

Peptide P7 specifically binds with basic fibroblast growth factor (bFGF) to inhibit the proliferation and invasion of numerous types of cancer cell. However, this effect has remained to be demonstrated in ovarian cancer-derived cell lines. In the present study, the protein P7 was used treat bFGF-stimulated SKOV3 epithelial ovarian cancer cells to explore the therapeutic potential of P7. An MTT and a scratch wound assay were used to respectively evaluate the proliferation and migration of bFGF-stimulated SKOV3 cells treated with P7. Reverse transcription-quantitative polymerase chain reaction analysis was used to detect the gene expression of urokinase-type plasminogen activator (uPA), as well as matrix metallopeptidase (MMP)-2 and −9, which have a role in cell migration/invasion. The morphology and proliferation of SKOV3 cells were not significantly affected by different concentrations of P7. However, P7 had an obvious inhibitory effect on the proliferation and migration of bFGF-stimulated SKOV3 cells. Treatment with P7 significantly lowered the gene expression of uPA, MMP-2 and MMP-9 compared with that in the control group. In conclusion, the present results suggested that P7, which, at least in part, acts through inhibition of bFGF, may have a potential therapeutic application in epithelial ovarian cancer.

Lnc00908 promotes the development of ovarian cancer by regulating microRNA-495-5p.

The aim of this study was to investigate whether lnc00908 could affect the proliferative and migratory behaviors of ovarian cancer (OC) cells by regulating microRNA-495-5p, thus participating in the development of OC. Quantitative Real Time-Polymerase Chain Reaction (QRT-PCR) was used to detect the expression levels of lnc00908 and microRNA-495-5p in OC tissues and normal ovarian tissues, as well as OC cell lines. The regulatory effects of lnc00908 and microRNA-495-5p on the proliferative and migration abilities of OC cells were detected by cell counting kit-8 (CCK-8) and transwell assay, respectively. The binding relationship between microRNA-495-5p and ANXA3, as well as miR-495-5p and lnc00908, was examined by luciferase reporter gene assay. Gain-of-function experiments were conducted to verify whether lnc00908 could affect the proliferative and migratory behaviors of OC cells by regulating microRNA-495-5p. Lnc00908 was highly expressed in OC tissues, and its expression was positively correlated with tumor stage. Overexpression of lnc00908 markedly promoted the proliferative and migratory abilities of SKOV3 and OVCAR cells. Luciferase reporter gene assay showed that lnc00908 could bind to microRNA-495-5p. However, microRNA-495-5p was significantly downregulated in OC tissues. Overexpression of microRNA-495-5p reversed the enhanced abilities of proliferation and migration in SKOV3 and OVCAR3 cells by lnc00908 overexpression. ANXA3 was a target gene of microRNA-495-5p. Moreover, overexpression of ANXA3 attenuated the inhibitory effect of miR-495-5p on the proliferative and migratory behaviors of SKOV3 and OVCAR3 cells. We found that the high expression of lnc00908 can promote the proliferation and migration abilities of OC cells through sponging microRNA-495-5p to regulate ANXA3 expression.

Knockdown of ARK5 expression suppresses invasion of ovarian cancer cells.

The aim of the current study was to investigate the effects and the molecular mechanisms of ARK5 in ovarian cancer cell invasion. The plasmid pGCsilencerU6/GFP/Neo‑RNAi‑ARK5 and the control vector with a scramble sequence were transfected into SKOV3 cells to establish ARK5‑deficient SKOV3 cells (siARK5/SKOV3) and a control cell line (Scr/SKOV3), respectively. Reverse transcription‑polymerase chain reaction (RT‑PCR) and Western blot analysis were used to determine the mRNA and protein expression levels of ARK5. Migration and invasion abilities of SKOV3 cells were determined in chemotaxis and invasion assays, respectively. The epidermal growth factor‑1 (EGF‑1)‑induced expression of matrix metallopeptidase (MMP)‑2 and MMP‑9, epithelial‑mesenchymal transition (EMT) and phosphorylation of mechanistic target of rapamycin kinase (mTOR) in siARK5/SKOV3 and Scr/SKOV3 cells were detected by western blot. RT‑PCR and western blot analyses demonstrated that the expression of ARK5 was significantly downregulated in siARK5/SKOV3 cells at the mRNA and protein levels (P<0.01). The migration and invasion abilities of siARK5/SKOV3 cells were markedly decreased compared with Scr/SKOV3 cells (P<0.01). In addition, the results demonstrated that EGF‑1‑induced expression of MMP‑2 and MMP‑9, EMT and phosphorylation of mTOR were suppressed in siARK5/SKOV3 cells as compared with Scr/SKOV3 cells (P<0.01). The current study demonstrated that ARK5 is a critical factor involved in SKOV3 cell invasion and ARK5 increases invasive potential by promoting EMT and activating the Akt‑mTOR‑MMPs pathway.

Icariin suppresses cell cycle transition and cell migration in ovarian cancer cells.

Ovarian cancer is the third most common type of gynecological tumor, in addition to being the most lethal. Cytoreductive surgery with chemotherapy is the standard treatment for ovarian cancer. It is necessary to identify novel chemotherapeutic methods, since current chemotherapy treatments are rarely effective for patients with advanced‑stage or recurrent ovarian cancer and may cause acute systemic toxicity. Icariin (ICA) is a prenylated flavonol glycoside derived from Herba Epimedii, a medicinal plant with a variety of pharmacological activities, including anticancer, antidiabetic and anti‑obesity effects. By analyzing cell viability, cell cycle and cell migration, the present study demonstrated that ICA inhibited the cell viability of the ovarian cancer cell line, SKOV3, and blocked cell cycle transition. ICA inhibited the expression of fuse binding protein 1 (FBP1), a critical regulator of proliferation and tumorigenesis through binding to the c‑Myc promoter, as well as β‑catenin, a key regulator in ovarian cancer initiation, metastasis, chemoresistance and recurrence. Furthermore, it was indicated that ICA inhibited the migration of SKOV3 cells. In accordance with our previous findings on high FBP1 expression in ovarian cancer, FBP1 was a potential target of ICA in ovarian cancer cells. Based on these results, the present study demonstrated that ICA may be a potential therapeutic agent for ovarian cancer treatment.

MicroRNA-16 inhibits migration and invasion via regulation of the Wnt/β-catenin signaling pathway in ovarian cancer.

As small non-coding RNA molecules, microRNAs (miRs) function in the regulation of tumorigenesis. Proliferation in ovarian cancer is considered to be associated with miR-16; however, the role of miR-16 in the migration and invasion of ovarian cancer cells remains unclear. The results of the present study demonstrated that miR-16 expression is downregulated in the ovarian cancer SKOV3 and OVCAR3 cell lines compared with that in normal ovarian epithelial cells (OECs). miR-16 overexpression inhibited the proliferation, migration and invasion of SKOV3 and OVCAR3 cells, and decreased the expression of matrix metallopeptidase (MMP)2 and MMP9. Additionally, miR-16 upregulated the expression of cadherin 1, an intercellular adhesion molecule, and downregulated the expression of some mesenchymal markers, including snail family transcriptional repressor 2, snail family transcriptional repressor 1, Vimentin, twist family BHLH transcription factor 1 and cadherin 2 in SKOV3 and OVCAR3 cells. Furthermore, it was indicated that miR-16 overexpression in SKOV3 and OVCAR3 cells resulted in a significant increase in anti-glycogen synthase kinase 3 β expression and a decrease in the expression of Wnt family member 3A, β-catenin, MYC proto-oncogene, BHLH transcription factor and cyclin D1 compared with the NC group. The results of the present study indicated that miR-16 exerts a suppressive effect on cell migration and invasion in ovarian cancer in vitro, through inactivation of the Wnt/β-catenin signaling pathway. The data suggest that miR-16 may be a potential therapeutic agent for the treatment and prevention of ovarian cancer.

UNBS5162 inhibits SKOV3 ovarian cancer cell proliferation by regulating the PI3K/AKT signalling pathway.

Ovarian cancer is the gynaecological malignancy with the highest mortality rate worldwide, and effective and safe therapeutic methods are limited. UNBS5162, a derivative of naphthalimide, has a clear inhibitory effect on the proliferation of various tumour cells in vitro and in vivo as a pan-antagonist of CXC chemokine ligand expression, but whether it serves a function in ovarian cancer remains unclear. The aim of the present study was to determine the effects of UNBS5162 on the proliferation, migration and invasion of ovarian cancer cells. The cell viability was detected using a Cell Counting Kit-8 (CCK-8) assay. The invasion and migration of SKOV3 cells were determined using Transwell assays. Cell apoptosis was determined using flow cytometry. The apoptosis-associated proteins and associated factors, such as phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signalling pathway members, were detected using western blot analysis. The CCK-8 assay revealed that SKOV3 cell viability was affected by UNBS5162 in a dose- and time-dependent manner. In Transwell assays, UNBS5162 inhibited cell invasion and migration. Furthermore, it was identified that UNBS5162 markedly increased the apoptosis rate of SKOV3 cells. Simultaneously, the expression of the anti-apoptotic protein B-cell lymphoma 2 (Bcl-2) was decreased and the expression of the pro-apoptotic proteins active caspase-3 and Bcl-2-associated X protein were increased in SKOV3 cells treated with UNBS5162. In addition, the expression levels of phospho (p-)AKT/total AKT, p-mammalian target of rapamycin (mTOR)/total mTOR, p-p70 S6 kinase (p70S6K)/total p70S6K and cyclin D1 were decreased in the UNBS5162-treated group. The results of the present study indicated that UNBS5162 inhibits proliferation, migration and invasion, and induces apoptosis in ovarian cancer cells, which may be regulated by the PI3K/AKT signalling pathway. These results suggest that UNBS5162 may be a potential novel drug for clinical ovarian cancer treatment.

SND1 acts upstream of SLUG to regulate the epithelial-mesenchymal transition (EMT) in SKOV3 cells.

Staphylococcal nuclease domain-containing protein 1 (SND1) has been reported as an oncoprotein in a variety of cancers involving multiple processes, including proliferation, angiogenesis, and metastasis. However, the mechanisms underlying metastasis remain largely unknown. Herein, by using the ovarian cancer cell line SKOV3, which has high metastasis ability, we showed that loss-of-function of SND1 dramatically suppressed the invasion and migration of SKOV3 cells. We then performed gene expression profiles and further verified (by use of quantitative PCR and Western blot analysis) that loss-of-function of SND1 resulted in up-regulation of epithelial markers, such as epithelial cadherin and claudin 1, and down-regulation of mesenchymal markers, including neural cadherin and vimentin. Moreover, we illustrated that SLUG, a key transcription factor implicated in epithelial-mesenchymal transition and metastasis, acts as an essential effector of the SND1-promoted epithelial-mesenchymal transition process via regulating N-CAD and VIM expression (or E-CAD and CLDN1). The underlying molecular mechanisms illustrated that SND1 regulates the gene transcriptional activation of SLUG by increasing chromatin accessibility through the recruitment of the acetyltransferases GCN5 and CBP/p300 to the SLUG promoter proximal region. Overall, SND1 was identified as a novel upstream regulator of SLUG, which plays important roles in regulating the E-CAD/N-CAD expression switch.-Xin, L., Zhao, R., Lei, J., Song, J., Yu, L., Gao, R., Ha, C., Ren, Y., Liu, X., Liu, Y., Yao, Z., Yang, J. SND1 acts upstream of SLUG to regulate the epithelial-mesenchymal transition (EMT) in SKOV3 cells.

MiR-122 inhibits epithelial mesenchymal transition by regulating P4HA1 in ovarian cancer cells.

Ovarian cancer is one of the most common gyneacologic malignancies, with high morbidity and high mortality. Hsa-miR-122-5p (miR-122) has been reported with tumor-suppressing roles in various cancers. In this study, miR-122 was overexpressed in ovarian cancer cells, and phenotypic experiments demonstrated that miR-122 inhibited migration and invasion in SKOV3 and OVCAR3 cells. MiR-122 also suppressed epithelial mesenchymal transition (EMT), evidenced by expression changes of E-cadherin, vimentin, matrix metalloproteinase (MMP)2, and MMP14. Prolyl-4-hydroxylase subunit alpha-1 (P4HA1) was identified as a target of miR-122, and downregulated by miR-122. MiR-122-induced the elevation of migration, invasion, and EMT were recovered by P4HA1. Additionally, miR-122 restrained the tumor metastasis of SKOV3 cells in peritoneal cavity of nude mice. In summary, we demonstrated that miR-122 inhibited migration, invasion, EMT, and metastasis in peritoneal cavity of ovarian cancer cells by targeting P4HA1 for the first time, which shed lights on the discovery of miR-122 and P4HA1 as possible potential diagnostic markers and therapeutic targets for ovarian cancer.

Integrated microarray meta-analysis identifies miRNA-27a as an oncogene in ovarian cancer by inhibiting FOXO1

AimsSurvival of ovarian cancer patients is generally poor, partly because most of them are already at an advanced stage when diagnosed. The purpose of this study was to screen prognostic miRNAs for ovarian cancer, and to explore the underlying mechanisms. Main methodsIntegrated meta-analysis of miRNA microarrays retrieved from public repositories was employed to identify clinically significant miRNAs involved in ovarian cancer. Targets of candidate miRNA were predicted using four online databases, and validated with dual luciferase assay. Loss and gain of function were performed to investigate the role of miR27a in the growth of ovarian cancer cell lines. Key findingsBased on cross-validation results in multiple datasets, we recognized hsa-miR-27a as an oncogenic molecular and a prognostic factor for ovarian cancer patients. Dual luciferase assay indicated tumor suppressor FOXO1 was a direct target of miR-27a. In addition, hsa-miR-27a could stimulate SKOV3 and A2780 cell proliferation and migration by regulating the expression of FOXO1. SignificanceIn summary, our results indicate that miR-27a can promote progression of ovarian cancer by mediating FOXO1. To our knowledge, this is the first study focusing on the role of miR-27a/FOXO1 axis using the microarray meta-analysis in ovarian cancer. Furthermore, inhibiting miR-27a expression may be a new strategy for the treatment of ovarian cancer.

Tumor-associated macrophages promote the metastasis of ovarian carcinoma cells by enhancing CXCL16/CXCR6 expression

This study investigated the underlying mechanism by which C-X-C motif chemokine ligand 16 (CXCL16)/C-X-C motif chemokine receptor 6 (CXCR6) signaling is activated by tumor-associated macrophages and assists in regulating the metastasis of ovarian carcinoma. Specimens of ovarian carcinoma tissue and adjacent tissue were collected from 20 ovarian carcinoma patients. Human THP-1 cells were induced to differentiate into macrophages, which were then co-cultured with SKOV3 cells and low concentrations of tumor necrosis factor-α (TNF-α) to simulate the inflammatory microenvironment of ovarian carcinoma. Additionally, small interfering RNA (siRNA) targeting CXCR6 was transfected into SKOV3 cells; after which, the levels of nuclear factor kappa B p65 (NF-κB p65) protein and phosphorylated PI3K and Akt were measured. The migration and invasion abilities of the SKOV3 cells were also tested. The levels of TNF-α, interluekin-6 (IL-6), NF-κB p65, CXCL16, and CXCR6 expression in the ovarian carcinoma tissues were higher than those in the precancerous tissues. CXCR6 expression was positively correlated with TNF-α, IL-6, and CXCL16 expression. Co-culture of SKOV3 cells with macrophages significantly promoted CXCL16, CXCR6, NF-κB, and p65 expression by the SKOV3 cells, increased their levels of phosphorylated PI3K and Akt, and increased the migration and invasion abilities of SKOV3 cells. Silencing of CXCR6 or blocking the PI3K/Akt signal pathway markedly attenuated the expression of NF-κB p65 and phosphorylation of PI3K and Akt, as well as the migration and invasion abilities of SKOV3 cells. These findings demonstrate that macrophages can promote the migration and invasion of ovarian carcinoma cells by affecting the CXCL16/CXCR6 pathway.

Abstract 495: Leptin facilitates ovarian cancer metastasis through miR-28-3p and matrix metalloproteinase-1 in the omental tumor microenvironment

Abstract Advanced stage serous ovarian cancers spread beyond the pelvis and preferentially metastasize to the omentum, which is mainly composed of adipose tissue, suggesting that the omental microenvironment is a favorable niche for ovarian cancer cells. By comparing the transcriptome profiles of microdissected ovarian cancer tissues from the primary ovarian site and from the omental site, we identified matrix metalloproteinase-1 (MMP-1) as one of the most significant genes that was up-regulated in the omental site compared with that in the primary site. To identify mediators secreted by the omental microenvironment that up-regulated MMP-1 in the omental site tumors, transcriptome profiling was performed on microdissected ovarian tumor associated adipose tissue in the omental site and the ovarian stromal tissue in the primary ovarian tumor site. We identified leptin as one of the top differentially expressed genes in ovarian tumor associated adipose tissue in the omental site, suggesting that it might up-regulate MMP-1 in the omental ovarian cancer. To further delineate the role of MMP-1 in mediating the effect of leptin on ovarian cancer cell metastasis, ovarian cancer SKOV3 and OVCA433 cells were treated with leptin. The results showed that leptin increased MMP-1 mRNA and protein expression as assessed by quantitative RT-PCR and Western blot analyses, respectively. Furthermore, we demonstrated that leptin induced both the cell migration and invasion potential of SKOV3 and OVCA433 cells, and the effects were abrogated by transfecting cells with MMP-1 siRNAs. In addition, using Ion Torrent next generation sequencing, we found that miR-28-3p is down-regulated in leptin-treated SKOV3 and OVCA433 cells compared to the untreated cells, suggesting that it may play a role in mediating the effect of leptin on MMP-1 expression and the subsequent ovarian cancer cell motility and invasion potential. In conclusion, leptin secreted by the adipocytes in ovarian cancer associated omental microenvironment may be important in facilitating the ovarian cancer cell metastasis through the miR-28-3p/MMP-1 pathway. Citation Format: Chi Lam Au Yeung, Ngai Na Co, Tsz-Lun Yeung, Samuel C. Mok. Leptin facilitates ovarian cancer metastasis through miR-28-3p and matrix metalloproteinase-1 in the omental tumor microenvironment [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 495.

Prucalopride Inhibits Proliferation of Ovarian Cancer Cells via Phosphatidylinositol 3-Kinase (PI3K) Signaling Pathway.

BackgroundOvarian cancer is the second most common malignant tumor of the female reproductive system and is the leading cause of death of gynecological malignancies, but at present there is no effective and safe therapy. There is no previously published report on the anti-cancer effect of prucalopride, which is a high-affinity 5-HT4 receptor. The aim of the present study was to determine whether prucalopride can inhibit proliferation of ovarian cancer cells.Material/MethodsThe cell viability was detected by use of the Cell Counting Kit-8 (CCK-8) assay. The invasion and migration of SKOV3 and OVCAR3 cells was detected by Transwell assay. The cell apoptosis was detected by apoptosis flow detection and Caspase-Glo 3/7 Assay Systems. The apoptosis-related proteins, autophagy marker proteins, and the related-factors of phosphatidylinositol 3-kinase (PI3K) were detected by Western blot.ResultsThe CCK-8 proliferation test showed that prucalopride inhibited the growth of ovarian cancer cell lines SKOV3 and OVCAR3. In the Transwell assay, prucalopride inhibited cell invasion and migration. Furthermore, we found the expression of anti-apoptotic protein Bcl-2 decreased, whereas the expression of pro-apoptotic protein Caspase3 and Bax increased in the SKOV3 cell line treated with prucalopride, as well as cleaved PARP. In addition, the expression of p-AKT, p-mTOR, and p70S6K decreased in the prucalopride-treated group, and the expression of autophagy marker protein LC3-II/I and Beclin1 significantly increased, whereas the expression of p62 protein decreased.ConclusionsThe present study reveals that in ovarian cancer cells, prucalopride inhibits proliferation, migration, and invasion, and induces apoptosis and autophagy, which may be regulated by the PI3K signaling pathway. These results suggest prucalopride has potential as a new drug for clinical ovarian cancer treatment.

The lncRNA TP73-AS1 promotes ovarian cancer cell proliferation and metastasis via modulation of MMP2 and MMP9.

Ovarian cancer is one of the most common gynecologic malignancy with poor prognosis. Recently, long noncoding RNAs (lncRNAs) have been identified as key regulators in cancer development. The current study investigated the role of lncRNA P73 antisense RNA 1T (TP73-AS1) in ovarian cancer. Quantitative real-time polymerase chain reaction determined the expression levels of TP-73AS1, matrix metallopeptidases (MMPs) messenger RNA. Cell proliferative ability, cell invasion, and migration were CCK-8 and colony formation, and transwell invasion and migration assays, respectively. The protein levels of matrix metallopeptidase 2 (MMP2) and MMP9 were measured by Western blot. TP73-AS1 was upregulated in the ovarian cancer tissues and ovarian cancer cells, and upregulation of TP73-AS1 was associated with poor prognosis. Knockdown of TP73-AS1 significantly suppressed cell proliferation, invasion, and migration of SKOV3 cells, and overexpression of TP73-AS1 promoted cell proliferation, invasion, and migration of OVCA429 cells. In addition, knockdown of TP73-AS1 suppressed the in vivo tumor growth. Tumor metastasis RT2 profiler polymerase chain reaction array showed that MMP2 and MMP9 was significantly upregulated by TP73-AS1 overexpression in ovarian cancer cells. TP73-AS1 overexpression enhanced the expression of MMP2 and MMP9 in ovarian cancer cells. Knockdown of MMP2 and MMP9 attenuated the effects of TP73-AS1 overexpression on cell invasion and migration. The clinical data showed that MMP2 and MMP9 were upregulated and positively correlated with TP73-AS1 expression in ovarian cancer tissues. Collectively, our results demonstrated the oncogenic role of TP73-AS1 in ovarian cancer, and targeting TP73-AS1 may represent a novel approach in battling against ovarian cancer.

Upregulation of KIF26B, Cell Migration and Proliferation of Human Ovarian Cancer Cell Lines In Vitro, and Patient Outcomes from Human Bioinformatic Analysis

BackgroundThe aim of this study was to assess the expression profile of the KIF26B gene, its effects on ovarian cancer cell behavior in vitro, the clinical prognostic value of expression of the KIF26B gene and associated signaling pathways, from bioinformatics data analysis.Material/MethodsThe Cancer Genome Atlas-Ovarian Cancer (TCGA-OV) database and the online Kaplan-Meier plotter for ovarian cancer were analyzed. Human ovarian cancer cell lines A2780 and SKOV3 were used to study the in vitro effects of KIF26B gene expression on cell proliferation and cell invasion. Genes that interacted with, co-localized to, and were co-expressed with the KIF26B gene (Pearson’s r ≥0.6) were identified and underwent Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis.ResultsIncreased expression of the KIF26B gene increased the proliferation and migration of A2780 and SKOV3 cells in vitro. KIF26B protein expression was upregulated in ovarian cancer tissue and was associated with lymphatic and venous invasion. TCGA-OV data analysis showed that increased expression of the KIF26B gene was significantly associated with reduced 3-year, 5-year, and 10-year overall survival (OS), progression-free survival (PFS), and post-progression survival (PPS). The genes that interacted with, co-localized to, and were co-expressed with KIF26B were enriched in several KEGG pathways; upregulation of the KIF26B gene was associated with TGF-β signaling pathway.ConclusionsUpregulation of KIF26B enhanced proliferation and migration of ovarian cancer cells in vitro. Bioinformatics analysis supported the association between increased expression of the KIF26B gene and reduced clinical outcome in patients with ovarian carcinoma.

MicroRNA-299-3p regulates proliferation, migration and invasion of human ovarian cancer cells by modulating the expression of OCT4

Ovarian cancer is among the most prevalent and lethal types of cancers in women. Several factors such as late diagnosis, unavailability of the reliable biomarkers, frequent relapses and dearth of efficient therapeutic targets form bottleneck in the treatment of ovarian cancers. In this study we investigated the potential of less studied miR-299-3p as the therapeutic target for the treatment of ovarian cancer. The results of the present investigation revealed that miR-299 is significantly upregulated in the ovarian cancers and suppression of its expression inhibits the proliferation by induction of apoptosis as well suppresses migration and invasion of the SKOV3 cancers cells. Further, OCT-4 was found to be putative target of miR-99-3p in ovarian cancer and inhibition of OCT-4 had similar effects as that of miR-299 inhibition on cell migration and invasion. Intriguingly, even overexpression of miR-299-3p could not rescue the effects of OCT-4 suppression on SKOV3 cell proliferation, migration and invasion. On contrary, overexpression of OCT-4 in SKOV3 cells transfected with miR-299-3p transfected could nullify the effects of miR-200-3p on proliferation, migration and invasion of the SKOV3 cells. Taken together, miR-299-3p regulated cell proliferation and metastasis by modulating the expression of OCT-4 and as such may prove to be an important therapeutic target.

Long non-coding RNA RP11-552M11.4 promotes cells proliferation, migration and invasion by targeting BRCA2 in ovarian cancer.

The present study aimed to investigate the effect of long non‐coding RNA (lncRNA) RP11‐552M11.4 on cell proliferation, apoptosis, migration and invasion as well as its targeting genes in epithelial ovarian cancer (EOC) cells. LncRNA RP11‐552M11.4 expression was detected in 67 tumor tissues and paired adjacent tissues obtained from EOC patients. lncRNA RP11‐552M11.4 mimic/inhibitor plasmids were transferred into ovarian cancer cells (SKOV3, A‐2780) and normal ovarian epithelial cells (IOSE80 cells). In addition, rescue experiment was carried out by transferring BRCA2 inhibitor&lncRNA RP11‐552M11.4 inhibitor plasmids into SKOV3 and A‐2780 cells. qPCR, western blot, CKK‐8, Annexin V/propidium iodide (AV/PI), wound‐healing and Matrigel invasion assays were carried out to detect RNA expression, protein expression, cell proliferation, apoptosis, migration, and invasion, respectively. LncRNA RP11‐552M11.4 expression was elevated in tumor tissues compared with paired adjacent tissues and correlated with higher pathological grade, International Federation of Gynecology and Obstetrics stage and worse overall survival in EOC patients. LncRNA RP11‐552M11.4 promoted SKOV3 cell proliferation, migration and invasion whereas it inhibited apoptosis. Rescue experiment and luciferase reporter assay showed that lncRNA RP11‐552M11.4 regulated SKOV3 cells functions through binding BRCA2. Further experiments in A‐2780 cells also validated that lncRNA RP11‐552M11.4 induced A‐2780 cell proliferation while repressing apoptosis by targeting BRCA2. In addition, upregulation of lncRNA RP11‐552M11.4 increased IOSE80 cell proliferation, migration and invasion while decreasing apoptosis. In conclusion, lncRNA RP11‐552M11.4 correlates with worse prognosis, and promotes cell proliferation, migration, invasion, and inhibits cell apoptosis by down‐regulating BRCA2 in EOC.