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Objective:To explore the effects of neural precursor cell-expressed developmentally down regulated 8(NEDD8) covalent modification on ovarian cancer cell proliferation and apoptosis,and the possible underlying mechanisms.Methods:Use different concentrations of MLN4924 (0,0.125,0.25,0.5 mol/L) and human ovarian cancer cell line SKOV3,the expression levels of PAR3, HER2,Neddylated-cullins,P-IκBα were detected by Western blot and the secretion of IL-6 was measured by ELISA after 4 h treatment.The cell proliferation was determined by CCK-8 staining and the cell cycle and apoptosis were analyzed by flow cytometry after 72 h treatment.Results:MLN4924 inhibits the proliferation of SKOV3 cells in a dose-dependent manner,which is associated with reduced IL-6 secretion.Western blot revealed that MLN4924 dose-dependently inhibits the neddylation of cullins(reduced neddylated-cullins) and induced the accumulation of P-IκBα,whereas the expression levels of PARs and HER2 remained largely unchanged.At the same time MLN4924 dose-dependently induced cell cycle prove at S phase and the formation of a large number of tetraploids.Furthermore,apoptosis increased with the dose of MLN4924.Conclusion:NEDD8 covalent modification specific inhibitor MLN4924 can significantly inhibit the proliferation of SKOV3 cells and induce cell cycle arrest and apoptosis.The above inhibitory effects may partially result from impaired P-IκBα degradation and IL-6 secretion.
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Objective The aim of this study was to investigate the expression of IL-6 induced by LPS in ovarian cancer and the effect of IL-6 on the expression of EZH2 mRNA and protein in ovarian cancer cell lines.MethodsCulture ovarian cancer cell lines, according to the different medium composition, establish the experimental group and the control group, detect the concentration of IL-6 in the supernatant of ovarian cancer cells by ELISA and the OD values of each group by MTT assay.Western blotting was used to detect the expression of EZH2 protein in ovarian cancer cells.The mRNA expression of EZH2 was detected by RT-PCR.ResultsThe expression of IL-6 in the supernatant of ovarian cancer cells was significantly higher than that in the control group (P<0.05).The proliferation of ovarian cancer cells was inhibited and the difference was statistically significant (P<0.05).The expression of EZH2 protein in ovarian cancer cells was significantly decreased, and there was statistical difference (P<0.05).The mRNA expression of EZH2 was significantly decreased, and there was statistical difference (P<0.05).ConclusionLPS could induce the expression of IL-6 and inhibit cell proliferation in ovarian cancer cells.IL-6 could inhibit the expression of EZH2.
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Objective To evaluate the effects of malignant ascites on the morphological characteristics and the proliferation and migration abilities of the tumor cell and ovarian cancer cell lines(SKOV3)in the ovarian cancerous ascites.Methods Tumor cells extracted from the ovarian cancerous ascites were cultured in vitro with DMEM high glucose culture medium,and ovarian cancer cell lines( SKOV3) were cultured in DMEM high glucose and DMEM high glucose with different proportion of malignant ascites.The morphological characteristics of the cells were observed by optical microscope and electron microscope respectively.Cell proliferation ability was de-tected by CCK kit;The effect of SKOV3 on the migration of ovarian cancer cell lines was measured by scratch test.Results The morphological characteristics of tumor cells and ovarian cancer cell lines( SKOV3) in ovarian cancer ascites were significantly different.The proliferation ability of tumor cells was decreased without the asci-tes.The proliferation and migration abilities of SKOV3 cultured in mixed culture medium were significantly im-proved compared with the cells cultured in high glucose medium.Conclusion The change of tumor cell morphol-ogy in ascites benefits its abilities of proliferation and migration.The malignant ascites promote the abilities of pro-liferation and migration of ovarian cancer cell line(SKOV3).
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Aim To study the growth inhibitory effect of the conjugate ( ovarian cancer specific targeting peptide and cisplatin, OSTP-DDP ) that targeting ovarian cancer cells A2780. Methods Using chemical method to syn-thesize OSTP-DDP, ovarian cancer cells A2780 were cul-tured in vitro, using CCK-8 method ( Cell Counting Kit-8) to detect the growth inhibitory effect of ovarian cancer A2780 cells, which were disposed by OSTP-DDP and DDP. Annexin V-FITC was used to detect the cycle and apoptosis effect of ovarian cancer A2780 cells which were disposed by OSTP-DDP and DDP. Results According to the mass spectrometry and the high performance liquid chromatography ( HPLC ) analysis, OSTP-DDP was proved to synthesize successfully. CCK-8 assay showed that both OSTP-DDP and DDP could play the growth in-hibitory effect and showed a concentration-dependent manner when cells were treated in different concentrations (10,20,40,80,160,320μmol·L-1 ) respectively after 24 h, 48 h, 72 h. And the effect of OSTP-DDP was stronger than DDP (P<0. 05), indicated OSTP-DDP had targeted cytostatic effect. The result of the flow cytometry showed that cell cycle was mostly arrested in G1 phase after 72h treated by OSTP-DDP and DDP, the inhibitory effect of OSTP-DDP was stronger than DDP (P<0. 05). The apop-tosis effect of OSTP-DDP was stronger than DDP ( P <0. 01),suggested that OSTP-DDP had a stronger targeting apoptosis-inducing effect. Conclusion OSTP-DDP has the targeting growth inhibitory effect on the ovarian cancer cell A2780, OSTP as a chemotherapeutic drug targeting vector has a great prospect to treat ovarian cancer.
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Objective To evaluate the effect of suberoylanilide hydroxamic acid(SAHA) or/and paclitaxel(PTX) on lethality and autophagy of human ovarian cancer OC3 cells,and to explore whether the combination of the two drugs has a synergistic function.Methods The morphology of OC3 cells was treated with SAHA and/or PTX, and then the morphology of treated OC3 cells was observed under an inverted microscope, cell proliferation was detected by MTT assay and autoph-agy was analyzed by AO/EB double staining assay.The synergistic effect of SAHA and/or PTX was analyzed by factorial design and gold formula method.Results After treatment with SAHA and/or PTX, the morphology of OC3 cells in the combination group ( SAHA+PTX) displayed significant morphological changes.OC3 cells became less adherent and refrac-tive than in other groups.Cell proliferation by MTT assay demonstrated that the growth inhibition rate of the combination groups was higher than in groups treated with SAHA or PTX respectively( P<0.05) .Furthermore, the synergistic effect af-ter treatment with a combination of SAHA with PTX was proved by the factorial design and gold formula method.The auto-phagy rate of the combined groups was significantly higher than in single treatment groups (P<0.05) by AO/EB double staining.Conclusion SAHA and PTX can inhibit the survival of OC3 cells and induce its autophagy.The two drugs have synergistic antitumor effects.
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Objective:To develop a biotin-streptavidin system (BAS)-mediated folate receptor (FR)-targeted quantum dot (QD) fluorescent probe and preliminarily validate the targeting ability and signal amplification effect of the probe. Methods: Streptavidin (SA) was covalently coupled with QD through the active ester method;the physical characteristics of the prepared QD-SA were veri-fied. Biotinylated folate was synthesized through the carrier bovine serum albumin using the same method and then reacted with QD-SA to form the special probe. The probe was used to identify SKOV3 cells and FR-negative A549 cells to verify its targeting speci-ficity. QD-SA was used as the contrast. SKOV3 cells were imaged using the BAS-mediated FR-targeted QD probe with a biotinylated folate incubation time of 1 or 4 h. Various reaction times were also tested between the probe and the QD-FA that was formed without BAS mediation. Results:The BAS-mediated FR-targeted QD probe specifically recognized FR-positive SKOV3 cells. The probe ob-tained higher fluorescent intensity after 4 h than after 1 h of biotinylated folate incubation. The BAS-mediated FR-targeted QD probe al-so had a stronger fluorescent signal than the QD-FA probe. Conclusion:The proposed probe presents a great potential in the early diag-nosis of ovarian cancer because of its high specificity and sensitivity.
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Objective To evaluate the effect of SAHA or/and PTX on survival and apoptosis of human paclitaxel-resist-ant ovarian cancer OC3/P cells, and explore whether the combination of two drugs has a synergistic effect .Methods The morphology of OC3/P cells in different drug-groups was observed by inverted microscope .Cell viability was evaluated by MTT assay.The apoptosis rate was analyzed by Annexin V-FITC/PI assay.Results The morphology change of OC 3/P cells treated with different drug was observed by inverted microscope , and the change in combination group was more signif-icant than one drug alone group .The result of cell survival measured by MTT assay showed that inhibition rate of combina -tion group was more higher than one drug alone group (P<0.05).The analysis of factorial design and gold formula method all proved that the two drugs had synergy .Further the result of flow cytometry showed that apoptosis rate in combination group was significantly higher than SAHA or PTX alone group (P<0.05).Conclusion SAHA and PTX can inhibit the survival and induce apoptosis of OC 3/P cells, and two drugs have synergistic antitumor effects .
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Objective To explore the mechanism of drug resistance of ovarian cancer cells to TRAIL-induced apoptosis.Methods We collected 3AO cells and CAOV3 cells,respectively,at 18,24,48 and 72 hour under 12.5,25,50 and 100 ng/mL concentrations of TRAIL.The rate of cell growth inhibition was checked by methyl thiazolyl tetrazolium (MTT)assay to evaluate the effect of TRAIL.Morphology of apoptotic cells was observed by TdT-mediated-dUTP nick end labeling (TUNEL).The apoptosis rate was detected by flow cytometry (FCM)and C-FLIP protein was determined by Western blotting.Results TRAIL inhibited the growth of 3AO and CAOV3 cells.The rate of growth inhibition at 24 hour was 28% in 3AO cells and 10% in CAOV3 cells.TRAIL induced apoptosis of cells.The apoptosis rate at 24 hour was 8.5% in 3AO cells,which was higher than 5.5% in CAOV3 cells.The expression level of C-FLIP protein was higher in CAOV3 cells than in 3AO cells.Conclusion C-FLIP protein is an important protein that regulates drug resistance of ovarian cancer cells to TRAIL-induced apoptosis.
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OBJECTIVE: To investigate the relationship between cisplatin resistance and histone deacetylase (HDAC) isoform overexpression in ovarian cancer cell lines. METHODS: Expression of four HDAC isoforms (HDAC 1, 2, 3, and 4) in two ovarian cancer cell lines, SKOV3 and OVCAR3, exposed to various concentrations of cisplatin was examined by western blot analyses. Cells were transfected with plasmid DNA of each HDAC. The overexpression of protein and mRNA of each HDAC was confirmed by western blot and reverse transcriptase-polymerase chain reaction analyses, respectively. The cell viability of the SKOV3 and OVCAR3 cells transfected with HDAC plasmid DNA was measured using the cell counting kit-8 assay after treatment with cisplatin. RESULTS: The 50% inhibitory concentration of the SKOV3 and OVCAR3 cells can be determined 15-24 hours after treatment with 15 microg/mL cisplatin. The expression level of acetylated histone 3 protein in SKOV3 cells increased after exposure to cisplatin. Compared with control cells at 24 hours after cisplatin exposure, the viability of SKOV3 cells overexpressing HDAC 1 and 3 increased by 15% and 13% (p<0.05), respectively. On the other hand, OVCAR3 cells that overexpressed HDAC 2 and 4 exhibited increased cell viability by 23% and 20% (p<0.05), respectively, compared with control cells 24 hours after exposure to cisplatin. CONCLUSION: In SKOV3 and OVCAR3 epithelial ovarian cancer cell lines, the correlation between HDAC overexpression and cisplatin resistance was confirmed. However, the specific HDAC isoform associated with resistance to cisplatin varied depending on the ovarian cancer cell line. These results may suggest that each HDAC isoform conveys cisplatin resistance via different mechanisms.
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Blotting, Western , Cell Count , Cell Line , Cell Survival , Cisplatin , DNA , Hand , Histone Deacetylases , Histones , Neoplasms, Glandular and Epithelial , Ovarian Neoplasms , Plasmids , Protein Isoforms , RNA, MessengerABSTRACT
OBJECTIVE: The aim of this study was to identify apoptosis-related genes of ovarian cancer cell lines following cisplatin treatment. METHODS: We used IC50 values and fluorescence-activated cell sorting analysis to compare cell death in 2 ovarian cancer cell lines, namely, SKOV-3 and OVCAR-3, upon treatment with cisplatin. Moreover, the change in transcriptional levels of apoptosis-associated genes was measured with a dendron-modified DNA microarray. RESULTS: The protein levels for the up-regulated genes in each cell line were validated to identify the molecules that may determine the cellular behavior of cisplatin resistance. Eight genes were over-expressed in the 2 cell lines. The cisplatin-induced up-regulation of DAD1 in transcriptional and protein levels contributed to the cisplatin resistance of OVCAR-3, and the up-regulation of FASTK and TNFRSF11A in SKOV-3 resulted in its higher sensitivity to cisplatin than that of OVCAR-3. CONCLUSION: In the present study, we have identified a set of genes responsible for apoptosis following cisplatin treatment in ovarian cancer cell lines. These genes may give information about the understanding of cisplatin-induced apoptosis in ovarian cancer.
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Apoptosis , Cell Death , Cell Line , Cisplatin , DNA , Flow Cytometry , Inhibitory Concentration 50 , Ovarian Neoplasms , RNA, Messenger , Up-RegulationABSTRACT
OBJECTIVE: Flavopiridol that inhibits cyclin-dependent kinase, can cause cell cycle arrest, induce apoptosis in human tumor cell lines. In the present study, we investigated apoptotic effects of flavopiridol and the underlying molecular mechanisms in human ovarian cancer cell lines. METHODS: We used TOV-21G and TOV-112D cell lines. The cell viability was tested by MTT assay and apoptosis was assessed by TUNEL assay and annexin-V binding. Western blot was used to examine apoptosis related protein levels. MAP kinase activity was analyzed by non-radioactive MAP kinase assay kit. RESULTS: Treatment of TOV-21G and TOV-112D cells with flavopiridol (50 nM to 1000 nM) led to a dose- and time-dependent inhibition of cell growth and survival. Dose-related induction of apoptosis was also observed in these cell lines. Flavopiridol (500 nM) induced striking decreases in the levels of the antiapoptic proteins Mcl-1, Bcl-X(L), and XIAP in both cell lines. In contrast, expression of Bax, Bcl-2, and AIF was not significantly influenced by flavopiridol. Although flavopiridol resulted in accumulation of p53 in both cells, flavopiridol mediated apoptosis was p53 independent because it occurred to the same degree in TOV-112D cells in which p53 was inactivated by mutation. Flavopiridol treatment resulted in enhanced cleavage of pro-caspase 9 and activation of caspase 3. Apoptosis was associated with suppression of ERK activity. CONCLUSION: Although the precise mechanisms of flavopiridol mediated cytotoxicity have not been fully defined, these data suggest that flavopiridol has activity against ovarian cancers in vitro and is worthy of continued clinical development in the treatment of ovarian cancer.
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Humans , Apoptosis , Blotting, Western , Caspase 3 , Cell Cycle Checkpoints , Cell Line , Cell Line, Tumor , Cell Survival , Flavonoids , In Situ Nick-End Labeling , Ovarian Neoplasms , Phosphotransferases , Piperidines , Proteins , Strikes, EmployeeABSTRACT
Objective To evaluate the inhibitory effect of endostatin on ovarian cancer cell line SKOV3 and to investigate the possible mechanism of the inhibition.Methods The effect of endostatin on SKOV3 cell proliferation was studied by means of MTT.The cell apoptosis was detected by transmission electronic microscopy.BCL-2 and BAX expressions were measured in SKOV3 cells treated by endostatin by immunocytochemistry,RT-PCR and Western blot.Results Endostatin inhibited SKOV3 cell proliferation(P
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OBJECTIVE: In order to explore Mullerian inhibiting substance (MIS) effects on the ovarian neoplasia, the expression and localization of the MIS type II receptor (MISR II), the growth inhibitory effects of MIS, and the underlying molecular mechanisms were investigated in the ovarian cancer cell lines. METHODS: Expression of MISR II were studied in SKOV-3, OVCAR-3, and OVCAR-8 cell lines by immunohistochemical staining. The antiproliferative effects of MIS in these cell lines were investigated by methylthiazoletetrazolium (MTT) assay, fluorescence-activated cell sorting (FACS) analysis, annexin-V-FITC binding, and western blot analysis. RESULTS: All cell lines showed strong specific staining for MISR II, although staining in OVCAR-8 cells was more intense than that in SKOV-3 and OVCAR-3. Treatment of OVCAR-8 cells with MIS led to a dose- and time-dependent inhibition of cell growth and survival was determined use by MTT assay. But OVCAR-3 cells exhibited growth inhibition at higher doses after 48 hours of treatment and SKOV-3 cells did not demonstrate response. Using FACS analysis, exposure of OVCAR-8 cells to MIS (71 nM) resulted in G1 arrest after 24 hours of treatment. This pattern was changed by time-dependent increase in the percentage of cells with a sub G0G1 DNA content, suggesting apoptosis, after 48 hours of treatment. These results suggested that cell death be preceded by cell cycle arrest. Time-related induction of apoptosis was also observed in this cell line as measured by annexin-V-FITC binding. In OVCAR-8 cells, the growth inhibitory effects of MIS were mediated through specific induction of CDKI p16 protein expression and via regulation of E2F1 in the absence of detectable levels of pRb. We estimated that OVCAR-3 cells were affected by MIS through p16-independent, alternative mechanistic pathways, since the growth inhibitory effects of MIS were minimal. SKOV-3 cells did not express p16 protein. CONCLUSION: We have demonstrated that ovarian cancer cells express the MISR II. Epithelial ovarian cancer cells respond to MIS by growth inhibition. Although the precise mechanisms of MIS mediated inhibition of ovarian cancer cell growth have not been fully defined, these data suggest that MIS has activity against ovarian cancers in vitro and may also be an effective targeted therapy for ovarian cancer.
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Humans , Anti-Mullerian Hormone , Apoptosis , Blotting, Western , Cell Cycle Checkpoints , Cell Death , Cell Line , DNA , Flow Cytometry , Immunohistochemistry , Ovarian NeoplasmsABSTRACT
OBJECTIVE: To evaluate the effects of both exogenous and endogenous osteopontin on normal and malignant ovarian epithelial cell growth, and on paclitaxel chemo-resistance. METHODS: The ovarian cancer cell line OV429, which showed low level of endogenous osteopontin and paclitaxel sensitive cell line OV420, which showed high level of endogenous osteopontin, and a normal ovarian epithelial (HOSE: Human ovarian surface epithelial) cells were treated with purified osteopontin. Furthermore, OV420 was treated with osteopontin siRNA alone or in combination with paclitaxel. Proliferation rates and cell cycle progression of treated cells were determined by the tetrazolium colorimetric (XTT) assay and FACS analysis, respectively. RESULTS: Exogenous osteopontin increased the proliferation rate of OV429 and OV420 but had negligible effect on normal HOSE. Ovarian cancer cell lines treated with siRNA showed significantly reduced the growth rates (P<0.05), and they were arrested in G2/M phase of the cell cycle. Furthermore, OV420 treated with paclitaxel in the presence of osteopontin siRNA showed significantly decreased the survival rate. CONCLUSION: Osteopontin promote cell growth in malignant but not in normal ovarian epithelial cells, and may confer paclitaxel-resistance by adhesion to each cell and minimized the cell surface which exposure to chemo-agents.
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Humans , Cell Cycle , Cell Line , Epithelial Cells , Osteopontin , Ovarian Neoplasms , Paclitaxel , RNA, Small Interfering , Survival RateABSTRACT
In solid tumor, there is a hypoxic region where oxygen supply is insufficient. In this study, we found that one of the quinolone antibiotics, levofloxacin, made a human ovarian cancer cells, SK-OV-3, resistant to hypoxia, even in the presence of a platinum-based anti-cancer therapeutic, carboplatin; when the cells (2 X 10(5) cells/12 well multi culture dish) were grown in no glucose medium (0 g/l) under hypoxia (1% O2), all the cells became dead after 24 hours of culture in the absence of levofloxacin and carboplatin, whereas the cells still survived, at least, until 36 hours of culture in the presence of levofloxacin (10-100 microgram/ml) alone or in combination with carboplatin. The results might have some implications in treating solid tumor; if cancer patients should be treated for infection with antibiotics, quinolone antibiotics can aggravate tumor by making cancer cells more resistant to hypoxia. This is also true even when a patient is treated with carboplatin. Therefore, the results strongly suggest that we should be careful in choosing antibiotics when they are used for cancer patients. In this regard, our work could be a new guideline in choosing antibiotics when antibiotics are applied for treating cancer patients.
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Humans , Hypoxia , Anti-Bacterial Agents , Carboplatin , Cell Survival , Glucose , Levofloxacin , Ovarian Neoplasms , OxygenABSTRACT
OBJECTIVE: The chemotherapeutic agent Cisplatin (cis-diammminedichloroplatinum (II)) is particularly effective against ovarian carcinoma, however, its clinical success is limited by recurrent drug resistant tumors. It is mandatory to reveal the mechanism of cisplatin resistance for the ovarian cancer treatment or prognosis. This study assessed the expression of p53, p16, PTEN, and c-myc genes with cisplatin treatment in human ovarian cancer cell lines; cisplatin-sensitive (A2780) and cisplatin-resistant (A2780/CP70) ovarian cancer cell line to elucidate the mechanism of cisplatin resistance. METHODS: Cytotoxic assay for cisplatin was performed in cisplatin-sensitive ovarian cancer cell line, A2780 and cisplatin-resistant ovarian cancer cell line, A2780/CP70. After cisplatin treatment, expression of p53, p16, PTEN, c-myc was analyzed by Western blot analysis. RESULTS: PTEN expression was significantly about 30% higher in A2780 than in A2780/CP70. p16 expression was defective in both cell lines. p53 and c-myc expression was no difference in cancer cell lines. After cisplatin treatment, the expression of p53, PTEN, c-myc genes increased 2-3 times in both cell lines. CONCLUSION: Relatively lower expression of PTEN was detected in A2780/CP70 suggesting that PTEN expression might play a role in the development of cisplatin resistance in ovarian cancer cell line, A2780/ CP70.
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Humans , Blotting, Western , Cell Line , Cisplatin , Genes, myc , Ovarian Neoplasms , PrognosisABSTRACT
AIM: To investigate the effects of synthetical glucocorticoid dexamethasone(Dex) on the activation of two members of mitogen-activated protein kinase (MAPK) family, extracellular signal-regulated protein kinase1/2(ERK1/2 ) and p38 MAPK (p38) in human ovarian cancer cell line HO-8910. METHODS: The activation of ERK1/2 and p38 was determined by Western blot. RESULTS: Inhibition of activation of ERK1 and ERK2 by 10 -7 mol/L Dex occurred at 5 min, with maximum up to 41% and 54% respectively at 30 min ( P
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PURPOSE: The growth regulatory effect of retinoid derivatives could be mediated by the transcriptional inactivation of AP-1 oncogenic transcription factor. By using ovarian cancer cell lines we were to investigate the cross-regulation mechanism between retinoids and AP-1. MATERIALS AND METHODS: Cell proliferation assays were performed in 4 ovarian cancer cells (A2774, PA-1, OVCAR-3, SKOV-3) by increasing the concentrations of all-trans retinoic acid (ATRA), 9-cis retinoic acid (9RA), 13-cis RA (13RA), 4-hydroxyphenyl retinamide (4-HPR). Transient transfection and CAT ELISA were done to determine the selective activity of each retinoid on the RAR (alpha, beta, gamma), RXR (alpha, beta, gamma). and the negative activity on AP-1 (c-Jun). RESULTS: Antiproliferative effect of 4-HPR (IC50; 0.7~2.7 micrometer) was more potent than those of other retinoid derivatives (IC50; 2.7~9.0 micrometer). To assess the anticancer mechanism, we examined the effect of 4-HPR on the transriptional activity of retinoic acid receptors (RAR/RXR) and of c-jun. Contrary to other retinoid derivatives that are active for RAR and RXR with some different levels, 4-HPR showed weak activity only for RARgamma. However, 4-HPR exerted the strongest suppression on AP-1 (c-Jun) activity. CONCLUSION: Based on our results showing much 4-HPR's potent antiproliferative activity coupled with the most effectively inhibiting activity on AP-1 and minimum activity on RA receptor (selective for RARgamma) than other retinoid derivatives, we suggest that 4-HPR may be a novel, and very effective anticancer drugs for ovarian cancer.
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Animals , Cats , Cell Line , Cell Proliferation , Enzyme-Linked Immunosorbent Assay , Fenretinide , Ovarian Neoplasms , Receptors, Retinoic Acid , Retinoid X Receptors , Retinoids , Transcription Factor AP-1 , Transcription Factors , Transfection , TretinoinABSTRACT
OBJECTIVE: Metastatic dissemination of epithelial ovarian carcinoma is thought to be mediated via tumor cell exfoliation into the peritoneal cavity, followed adhesion to and invasion through the mesothelium which overlies the contents of the peritoneal cavity. MMP-2 is secreted as a zymogen, the activation of which has been associated with metastatic progression in human ovarian cancer cell lines. METHODS: We have utilized short-term cultures to analyze the effect of specific extracellular matrix proteins, type I collagen. RESULTS: Culturing Caov-4 ovarian cell line on type I collagen led to a significant increase in conversion of the MMP-2,72kD to the MMP-2,66kD, and MT-MMP expression. MT-MMP expression correlates with expression and activation of MMP-2 during malignant progression. Altered MT-MMP expression in ovarian cell lines might contribute to MMP-2 activation, which facilitates invasion of these tumors. CONCLUSION: In summary, we found increased expression of MT-MMP that correlated with increased level of activated MMP-2 and cellular counts in chemoinvasion assay in Caov-3 cell line. But no significant increases in Skov-4 cell line on type I collagen. Conclusion: These data suggest that type I collagen induces MMP-2 activation in part by up-regulation of MT-MMP expression but has a more complicated mode of action involving additional processes.
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Humans , Cell Line , Collagen Type I , Epithelium , Extracellular Matrix Proteins , Matrix Metalloproteinases , Membranes , Ovarian Neoplasms , Peritoneal Cavity , Up-RegulationABSTRACT
Purpose:To investigate whether transforming growth factor ?1 (TGF ?1) pathway is involved in the mechanism of dexamethasone (Dex) mediated proliferation inhibition in ovarian cancer cell line HO 8910.Methods:To analyse cell proliferation and cell cycle distribution by cell counts and flow cytometric analysis, respectively ; to determine the expression levels of TGF ?1 and its two receptors, T?R Ⅰ and T?R Ⅱ , by quantative RT PCR, ELISA and(or) immunocytochemistry methods.Results:Dex induced a G 0 /G 1 cell cycle arrest in HO 8910 cells, and it up regulated T?R Ⅱ expression in a concentration dependent manner.The level of T?R Ⅱ mRNA was the highest after treatment with Dex for 8 hours, with 1.4 fold more than that of control at concentration of 10 -7 mol/L ( P