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Objective: To investigate the effect of silencing sirtuin 3 (Sirt3) on the apoptosis of human ovarian cancer SKOV3 cells induced by resveratrol (Res), and to explore its mechanism of promoting apoptosis. Methods: The human ovarian cancer SKOC3 cells were cultured with different concentrations 0, 2. 5, 5. 0, 10.0, 20.0, 40.0 and 80.0 mg · L-1) of Res for 24 h. The survival rate of cells was measured by MTT assay. The SKOV3 cells were randomly divided into control group, Sirt3 inhibitory 3-1H-1, 2, 3-triazol-4-yl) pyridine 3-TYP group, Res group and 3-TYP+Res group. After 24 h of culture, the inhibitory rates of proliferation of the cells in various groups were detected by MTT assay; the nuclei were stained with Hoechst 33342, and the morphorgy nucleus was observed by laser confocal microscope; reactive oxygen species (ROS) probe was used to detect the intracellular ROS levels; Western blotting method was used to detect the expression levels of Sirt3, Bax, Bcl-2 and cleaved caspase-3 proteins in the cells in various groups. Results: The results of MTT assay showed that the survival rates of SKOV3 cells were significantly decreased with the increase of concentration of Res, and the median inhibitory concentration (IC50) was 42. 73 mg · L-1. Compared with control group, the inhibitory rates of proliferation of cells in Res group and 3-TYP+Res group were significantly decreased (P0.05); the protein expression levels of Sirt3 and Bcl-2 proteins in Res group were significantly decreased (P< 0.05), and the expression levels of Bax and cleaved caspase-3 proteins were significantly increased (P<0.05). Compared with Res group, the expression levels of Bax and cleaved caspase-3 proteins in 3-TYP + Res group were significantly increased (P<0.05), and the expression levels of Bcl-2 and Sirt3 proteins in 3-TYP+Res group were significantly decreased (P<0.05). Conclusion: Res can induce the apoptosis of SKOV3 cells, and the inhibition of Sirt3 expression by 3-TYP can enhance the effect of Res.
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Objective To investigate the inhibitory effect of Xiaoaiping injection (XAP) combined with paclitaxel (PTX) on human ovarian cancer SK-OV-3 cells .Methods In vitro anti-proliferation activity study of XAP combined with PTX on hu-man ovarian cancer SK-OV-3 cells was performed using optical microscope and MTT assay .Human ovarian cancer SK-OV-3 cells were treated with PTX ,XAP ,PTX combined XAP or vehicle control .Each group of cells was treated with drugs for 24 h or 48 h .SK-OV-3 cell morphology was observed with optical microscope .MTT assay was used to detect the A value and cell vi-ability was calculated .In vivo effect of XAP combined with PTX on the growth of SK-OV-3 cells was determined in nude mice . In our study ,thirty-six mice were randomly divided into six groups :G1 (NS) ,G2 (PTX ,10 mg/kg) ,G3 (XAP ,20 ml/kg) , G4 (XAP ,50 ml/kg) ,G5 (PTX 10 mg/kg+XAP 20 ml/kg) and G6 (PTX 10 mg/kg+XAP 50 ml/kg) .Animals were trea-ted for 18 days .Body weight ,tumor volume and tumor inhibition rate were recorded and calculated .The results were analyzed by the SPSS 19 .0 software .Results In vitro study showed that SK-OV-3 cell viability decreased significantly in PTX com-bined XAP group compared to PTX group or XAP group ,in a time and dose-dependent manner .In vivo study showed that the combination of PTX and XAP resulted in decreased tumor weight significantly compared to the control or the PTX alone . Conclusion The combination of XAP and paclitaxel exhibited a synergistic effect both in vitro and in vivo in nude mouse tumor xenograft model .
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Inflammation has recently been implicated in cancer formation and progression. As tissue transglutaminase (TG2) has been associated with both inflammatory signaling and tumor cell behavior, we propose that TG2 may be an important link inducing interleukin-6 (IL-6)-mediated cancer cell aggressiveness, including cancer stem cell-like characteristics and distant hematogenous metastasis. We evaluated the effect of differential TG2 and IL-6 expression on in vivo distant metastasis of human ovarian cancer cells. IL-6 production in human ovarian cancer cells was dependent on their TG2 expression levels. The size and efficiency of tumor sphere formation were correlated with TG2 expression levels and were dependent on TG2-mediated IL-6 secretion in human ovarian cancer cells. Primary tumor growth and propagation in the peritoneum and distant hematogenous metastasis into the liver and lung were also dependent on TG2 and downstream IL-6 expression levels in human ovarian cancer cells. In this report, we provide evidence that TG2 is an important link in IL-6-mediated tumor cell aggressiveness, and that TG2 and downstream IL-6 could be important mediators of distant hematogenous metastasis of human ovarian cancer cells. Intervention specific to TG2 and/or downstream IL-6 in ovarian cancer cells could provide a promising means to control tumor metastasis.
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Humans , Axis, Cervical Vertebra , Inflammation , Interleukin-6 , Liver , Lung , Neoplasm Metastasis , Ovarian Neoplasms , PeritoneumABSTRACT
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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To construct a recombinant adenovirus containing CDglyES fusion gene, which can directly inhibit human ovarian cancer cell and indirectly inhibit vascular endothelial cell growth. Methods:We constructed prAdCDglyES using a homolo-gous recombination method in bacteria. The prAdCDglyES was transfected to 293 packaging cells using liposome, in which rAdCDgly-ES was packaged and amplified. MTT was used to observe the proliferation inhibition effect of rAdCDglyES on human ovarian cancer cells and the growth inhibition effect of expressing products of rAdCDglyES on ECV-304. Results:The titer of rAdCDglyES was 1 × 1013.3 TCID50/L, whereas the inhibition rate on human ovarian cancer cell SKOV-3 was (83.1±6.3)%. This result is significantly different from the control rAd-LacZ, which had an inhibition rate of (24.1 ± 13.2)% (P<0.01). The concentrated culture supernatant from cells transfected with rAdCDglyES can inhibit ECV-304 cell proliferation at a rate of (78.7 ± 1.6)%. This rate is significantly different com-pared with that of the control with the same concentration of culture supernatant from cells transfected with rAd-CD, with an effect on ECV-304 cell shown by an inhibition rate of (23.9 ± 9.7)%(P<0.01). Conclusion:The results showed that the recombinant adenovirus rAdCDglyES could inhibit human ovarian cancer cells directly and indirectly.
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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 study the effect of Xiaoaiping injection on Caov-3 human ovarian cancer cells and its mechanisms. Methods After treatment with Xiaoaiping injection, viability of Caov-3 cells determined by MTT method. Phase contrast microscopy was used to observed the morphological changes of Caov-3 cells. Cell cycle was assessed by FACS. Cell signaling pathway protein-Akt and pAkt, and cell cycle associated protein-p27 were measured by western blot. Results Xiaoaiping injection inhibited the growth of Caov-3 human ovarian cancer cells in a dose and time dependent manner. Xiaoaiping injection induced G0/G1 phase arrest of Caov-3 cells, accompanied by pAkt down-regulation and p27 up-regulation. Conclusion Xiaoaiping injection can inhibit the proliferation of Caov-3 human ovarian cancer cells by inhibiting PI3K/Akt signaling pathway.
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Objective To discuss the impact of zedoary turmerie oil(ZTO) injection over oncosis index and Bcl-2 expression of human ovarian cancer SKOV3 cell.Methods After ZTO injection acted on human ovarian cancer SKOV3 cells,the changes in the nucleus were observed by fluorescence microscope,oncosis index was counted by projection electron microscope,the situation of cell DNA breakage was observed by agarose gel electrophoresis,and cell Bcl-2 gene expression was observed by immunohistochemical method.Results After treated by ZTO injection for 48 hours,human ovarian cancer SKOV3 cells shows that oncosis cell was swelling in fluorescence microscope,the volume increased,the membrane area narrowed,the nuclear chromatin expansed.The oncosis index increased with dose-effect relationship,DNA electrophoresis showed diffuse type and Bcl-2 gene expression was down-regulated.Conclusion ZTO injection can increase oncosis index of human ovarian cancer SKOV3 cell with the concentration,and lower Bcl-2 gene expression,which may be one of the mechanisms of ZTO injection caused oncosis of SKOV3 cell.
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Objective To investigate the effect of caspase-2 and caspase-3 in cisplatin-induced apoptosis in cisplatin-resistant and-sensitive human ovarian cancer cell lines, A2780/DDP and A2780. Methods A2780 and A2780/DDP cells were incubated with various doses(0,5,10,20,40?M) of cisplatin for 24,48 and 72 hours, respectively. The protein expressions of caspase-2 and caspase-3 in the ovarian cancer cell lines were detected by immunohistochemical technology. The apoptotic rate of the ovarian cancer cell lines was determined by TUNEL. Results After cisplatin treated, apoptotic rates of both 2780 and A2780/DDP cell lines increased in time-and dose-dependent manners (P
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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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0.05).Conclusion:The decreased expressions of IL-18 and ICE in local ovarian cancer Tissues should have certain relation to the occurrence and development of ovarian cancer.
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Objective: To investigate reversal of drug resistance in human ovarian cancer cells by c-jun antisense. Methods: A c-jun antisense was applied to treat resistant and sensitive A2780 cell lines, and to observe the expression levels of c-jun, ?-GCS and GSH in two cell lines. Results: c-jun antisense inhibit c-jun gene expression in resistance cell lines . The mRNA level of the key enzyme in GSH synthesis, ?-glutamyl cysteine synthetase, was also reduced. The GSH content in resistant cells was dropped about 75 % . MTT analysis show that the resistant cells IC_(50) to cisplatin was dropped from 40 ?mol/L to 1.0 ?mol/L after a c-jun antisense treatment. No significant effect was observed in senstive cells (0.2 ?mol/L). Conclusion: A c-jun antisense can inhibit its gene expression in cells, and GSH synthesis in resistant cell was also inhibited. The resistant cells could be reversed to the level of sensitive cells.
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Objective To investigate the morphological changes of mitochondria and the expression of bcl-2 in the apoptotic SKOV3 and 3AO cells induced by arsenic trioxide (As2O3). Methods Light and electron microscopy, flow cytometry analysis, immunofluorescence flow cytometry analysis were used to detect the apoptotic cells, ultrastructural alteration of mitochondria, and the changes of mitochondrial transmembrane potentials (??m). Results As2O3 induced apoptosis of ovarian cancer cell lines was in a dose dependent manner and various in different cell lines. As2O3 also made a decrease of ??m in SKOV3 and 3AO cells in a dose independent fashion. Electron microscopy indicated that the mitochondria showed swollen, balloon-like appearance and outer membrane disrupted 72 h after As2O3 treatment. Expression of bcl-2 was down-regulated in SKOV3 and 3AO cells after As2O3 treatment. Conclusion The reduce of ??m and down-regulation of bcl-2 may play the key roles in the process of As2O3-induced apoptosis.
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Objective:To explore the effect of lipofectin-c-erbB2 antisense oligodexynucleotides on radiosensitivity of human ovarian cancer cell line.Methods:The expression of c-erbB2 was detected by means of RT-PCR;cellular response to irradiation was evaluated by MTT test and the colony forming assay.Results:Lipofectin-c-erbB2 antisense oligodexynucleotides(AS-ODN) could suppress the expression of c-erbB2,and significantly decreased the survival fraction and colony forming rate of human ovarian cancer cells after ionizing irradiation( P 0.05).Conclusion:c-erbB2 antisense oligodexynucleotides sensitize the SKOV3 to ionizing irradiation through decreasing the expression of c-erbB2,which might be the result of the fact that c-erbB2 antisense oligodexynucleotides inhibit the cellular signal transduction pathway relating to the radiation-resistant phenotype.
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Objective To investicate the effect of apoptosis of human ovarian cancer cell line SKOV_(3) and 3AO exposed to arsenic trioxide on telomerase activity and its mechanisms.Methods The human ovarian cancer cell line SKOV3 and 3OA were treated with arsenic trioxide of different concentration for 12,24,72 h.Cell morphology,PCR-ELISA,RT-PCR were adopted to detect the cell apoptosis and telomerase activity and the expression of human telomerase catalytic subunit(hTERT).Results Arsenic trixide could induce apoptisis of ovarian cancer cell lines,but there exists difference in drug concentration,type of cell line.A dose and time-dependent decline of telomerase activity after SKOV_(3) and 3AO cells exposed to arsenic trixide,meanwhile hTERT mRNA was down-regulated.Conclusion Telomerase activity and hTERT mRNA expression have close relationship and play an important role in arsenic trioxide inducing SKOV_(3) and 3AO cells apoptosis.