Lidocaine Inhibits Ovarian Cancer Cell Proliferation and Induces Apoptosis: An In Vitro Study

2020 ◽  
Vol 10 (5) ◽  
pp. 724-729
Author(s):  
Yaping Xu ◽  
Xiaoqin Fang ◽  
Xianjiang Wei
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Protein Expression ◽  
Western Blot ◽  
Cancer Cell ◽  
Blot Analysis ◽  
Group Treatment ◽  
Western Blot Analysis ◽  
Ovarian Cancer Cell ◽  
Control Group

Objective: The present study aimed to explore the effects and related mechanism of lidocaine on human ovarian cancer cell lines. Methods: Human ovarian cancer cell lines (SKOV3 and ES-2) were treated with different concentrations of lidocaine for different time. We treated SKOV3 and ES-2 cells using lidocaine then used MTT assay and flow cytometry to detect the cell proliferation and cell apoptosis. In addition, we used western blot analysis to explore the protein expression of Bax and Bcl-2 in SKOV3 and ES-2 cells. Western blot analysis and qRT-PCR were performed for the detection of EMT markers (E-cadherin, N-cadherin). The protein expression levels of TRAF3 and p-p65 in SKOV3 and ES-2 cells were determined by Western blot analysis. Results: Compared to the control group, 0.5, 1, 5, and 10 mM of lidocaine significantly inhibited ovarian cancer cell proliferation at different time points, while 0.1 mM of lidocaine had no significant effect. 1, 5 mM of lidocaine induced the cell apoptosis, and observably reduced expression of Bcl-2 protein, but improved Bax expression markedly compared with the control group. Treatment of lidocaine increased E-cadherin expression, but decreased N-cadherin expression when compared with control group. Treatment of lidocaine increased TRAF3 protein expression, but decreased p-p65 protein expression in ES-2 and SKOV3 cells. Conclusion: We demonstrated that lidocaine inhibited cell proliferation, induced apoptosis, and inhibited EMT in ovarian cancer cells via regulating TRAF3/NF-κB pathway.

scholarly journals GnRH-II receptor-like antigenicity in human placenta and in cancers of the human reproductive organs

2005 ◽  
Vol 153 (4) ◽  
pp. 605-612 ◽  
Author(s):  
Nicola Eicke ◽  
Andreas R Günthert ◽  
Volker Viereck ◽  
Doreen Siebold ◽  
Martin Béhé ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cell Membrane ◽  
Western Blot ◽  
Cell Lines ◽  
Cancer Cell ◽  
Human Placenta ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Ovarian Cancer Cell ◽  
Ovarian Cancer Cell Lines

We have recently demonstrated that the antiproliferative activity of GnRH-II on human endometrial and ovarian cancer cell lines is not mediated through the GnRH-I receptor. A functional receptor for human GnRH-II has not yet been identified. In this study, we have generated a polyclonal antiserum to the putative human GnRH-II receptor using a peptide (YSPTMLTEVPPC) corresponding to the third extracellular domain coupled to keyhole limpet haemocyanin via the Cys residue. A database search showed no identical peptide sequences in any other human gene. To avoid cross-reactions against two similar amino acid sequences the antiserum was pre-absorbed using these peptides. Immune histological sections of human placenta and human endometrial, ovarian and prostate cancers using rabbit anti-human GnRH-II receptor antiserum showed GnRH-II receptor-like staining. Western blot analysis of cell membrane preparations of human endometrial and ovarian cancer cell lines yielded a band at approximately 43 kDa whereas Western blot analysis of cell membrane preparations of ovaries obtained from the marmoset monkey (Callithrix jacchus) yielded a band at approximately 54 kDa. To identify the GnRH-II receptor-like antigen we used the photo-affinity labelling technique. Photochemical reaction of 125I-labelled (4-azidobenzoyl)-N-hydroxysuccinimide-[d-Lys6]-GnRH-II (10−9 M) with cell membrane preparations of human endometrial and ovarian cancer cells yielded a band at approximately 43 kDa. In competition experiments, the GnRH-I agonist Triptorelin (10−7 M) showed a weak decrease of 125I-labelled (4-azidobenzoyl)-N-hydroxysuccinimide-[d-Lys6]-GnRH-II binding to its binding site. The GnRH-I antagonist Cetrorelix (10−7 M) showed a clearly stronger decrease, whereas GnRH-II agonist [d-Lys6]-GnRH-II (10−7 M) was the most potent competitor. Western blot analysis of the same gel using rabbit anti-human GnRH-II receptor antiserum identified this band as GnRH-II receptor-like antigen.

TCP1 Regulates PI3K/AKT/mTOR Signalling Pathway to Promote Ovarian Cancer Cell Proliferation

2020 ◽  
Author(s):  
Huixi Weng ◽  
Xiushan Feng ◽  
Yu Lan ◽  
Zhiqun Zheng
Keyword(s):  
Ovarian Cancer ◽  
Protein Expression ◽  
Western Blot ◽  
Epithelial Ovarian Cancer ◽  
Cancer Cell ◽  
Ovarian Cancer Cell ◽  
Signalling Pathway ◽  
Migration And Invasion ◽  
Benign Ovarian Tumours ◽  
Pi3k Signalling

Abstract ObjectiveTCP1 is one of the eight subunits of the TCP1 ring complex (TRiC) or the multi-protein mammalian cytosolic chaperone complex. TRiC participates in protein folding and regulates the expression of multiple signalling proteins and cytoskeletal components in cells. Although the clinical importance of its subunits has been clarified in various carcinomas, the function of TCP1 in ovarian cancer (OC) remains unclear. We aimed to identify the association between the expression of TCP1 and epithelial ovarian cancer (EOC) development and patients’ prognosis, and explore the underlying mechanisms of TCP1 on the tumour progression of ovarian cancer cells.MethodsTCP1 protein expression was tested in the various ovarian tissues by immunohistochemistry (IHC), and the correlation between TCP1 expression and clinical physiologic or pathologic parameters of EOC patients was analyzed in this study. The relationship between TCP1 expression and ovarian cancer patients’ prognosis was collected and analyzed using the Kaplan-Meier (KM) Plotter online database. Then, the expression levels of TCP1 was tested in different OC cell lines by Western blot. Furthermore, a model using ovarian cancer cell line A2780 was constructed for studying the functions of TCP1 in human EOC cell growth, migration, and invasion. Finally, possible regulated signalling pathways were discussed.ResultsTCP1 protein expression in ovarian cancer or borderline tissue was significantly higher compared to that in benign ovarian tumours and normal ovarian tissue. The upregulated expression of TCP1 in ovarian cancer was positively associated with and the differentiation grade and FIGO stage, which predicted poor clinical outcomes. Compared with IOSE-80 cells, TCP1 protein was overexpressed in the A2780 cells. TCP1 knockdown using shRNA lentivirus inhibited cell viability in A2780 cells. Western blot showed that the phosphatidylinositol-3 kinase (PI3K) signalling pathway was activated in the tumour invasion of EOC driven by TCP1. ConclusionThe protein level of TCP1 is overexpressed in aggressive histologic types of epithelial ovarian cancer. Upregulated TCP1 is correlated with poor prognosis of patients and TCP1 may serve as a novel prognostic biomarker. The mechanism of cancer progression promoted by TCP1 upregulation may be linked to the PI3K signalling pathway activation and TCP1 may serve as a novel target for ovarian cancer treatment.

Novel method for predicting chemoresistance to paclitaxel in epithelial ovarian cancer patients

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 15007-15007
Author(s):  
D. Silasi ◽  
T. J. Rutherford ◽  
P. E. Schwartz ◽  
R. Chen ◽  
A. Alvero ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Protein Expression ◽  
Western Blot ◽  
Epithelial Ovarian Cancer ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Laser Capture Microdissection ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Laser Capture

15007 Background: No available test exists to guide the selection of effective chemotherapeutic regimen in recurrent ovarian cancer. Preliminary studies in our lab have identified a protein, MyD88, a major component in the inflammatory pathway, to be highly expressed in epithelial ovarian cancer cells that exhibit primary or acquired Paclitaxel chemoresistance. The objective of this study was to develop a sensitive approach that can detect expression of MyD88 in ovarian cancer tissue. We report the development of a test based on Laser capture microdissection that allows detection of MyD88 in a 6000 cell sample. Methods: Tumor tissue was obtained at surgery from epithelial ovarian cancer patients and snap-frozen in liquid nitrogen. Eight micron sections were prepared on polyethylene covered glass slides and tumor cells were dissected with a Laser capture microdissection system. Protein expression was detected by Western blot analysis. Results: Protein expression was detected by Western blot analysis in 1000 microdissected cells. An inverse correlation was observed between MyD88 expression in tumor cells and clinical response to Paclitaxel. Furthermore, this method allows the isolation of CD-45 positive cells from the tumor and analysis of their protein expression. Conclusions: We describe for the first time a method that will allow us to predict chemoresistance. Laser capture microdissection is a powerful technique that can be used to study the protein profile of each of the cellular components present in the tumor microenvironment. This technique will facilitate our understanding of the proteins necessary for tumor growth and may help to identify novel markers or potential protein targets. No significant financial relationships to disclose.

Protein Microarray Analysis of Nasal Polyps from Aspirin-Sensitive and Aspirin-Tolerant Patients with Chronic Rhinosinusitis

2009 ◽  
Vol 23 (3) ◽  
pp. 268-272 ◽  
Author(s):  
Kelly A. Zander ◽  
Milene T. Saavedra ◽  
James West ◽  
Victor Scapa ◽  
Linda Sanders ◽  
...  
Keyword(s):  
Protein Expression ◽  
Western Blot ◽  
Chronic Rhinosinusitis ◽  
Nasal Polyp ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Nasal Polyps ◽  
Protein Microarray ◽  
Control Group ◽  
Microarray Technology

Background The purpose of this study was to apply protein microarray technology to the study of sinonasal tissue and to identify differential protein expression in nasal polyps from aspirin-sensitive (AS) versus aspirin-tolerant (AT) patients with chronic rhinosinusitis (CRS) and CRS with nasal polyps (CRSwNPs). Methods Nasal polyp specimens were prospectively obtained from two groups of patients with CRSwNP. The test group (AS) consisted of five patients that were diagnosed with CRSwNP and intolerance to aspirin based on medical history and physical exam. The control group (AT) consisted of four AT patients with CRSwNP. Protein was extracted and labeled from harvested polyps and the Sigma Panorama Antibody Microarray–Cell Signaling Kit was used to identify differences in protein expression between the two polyp groups. Western blot analysis was used to validate the results of the protein microarray. Results The protein microarray showed a greater than twofold change in expression of both beta-adaptin and heat shock protein 70 (HSP70). Western blot analysis confirmed up-regulation of beta-adaptin and HSP70 in nasal polyp tissue from AS patients. Conclusion Pooled samples of AS and AT nasal polyps evaluated by protein microarray show distinct protein expression profiles in the stress response and receptor-mediated endocytosis pathways. This study establishes the successful application of protein microarray technology to study nasal polyposis, which in turn can be validated by Western blot analysis.

scholarly journals Ethanol Extracts of Solanum lyratum Thunb Regulate Ovarian Cancer Cell Proliferation, Apoptosis, and Epithelial-to-Mesenchymal Transition (EMT) via the ROS-Mediated p53 Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-16
Author(s):  
Chen Zhang ◽  
Zheming Li ◽  
Jie Wang ◽  
Xuelu Jiang ◽  
Mengting Xia ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Western Blot ◽  
Cancer Cell ◽  
Ovarian Cancer Cell ◽  
Ovarian Cancer Cells ◽  
Cancer Cell Proliferation ◽  
Anticancer Effects ◽  
Skov3 Cells

Ovarian cancer is a type of common gynecological tumors with high incidence and poor survival. The anticancer effects of the traditional Chinese medicine Solanum lyratum Thunb (SLT) have been intensively investigated in various cancers but in ovarian cancer is rare. The current study is aimed at investigating the effect of SLT on ovarian cancer cells. Lactate dehydrogenase (LDH) and MTT assays indicated that SLT concentrations of 0.25 and 0.5 μg/mL were not cytotoxic and had significant inhibitory effects on the cell viabilities of A2780 and SKOV3 cells, hence were used for subsequent experiments. Flow cytometric and western blot analysis revealed that SLT effectively suppressed ovarian cancer cell proliferation via inducing cell cycle arrest and increasing apoptosis. Cell cycle and apoptosis-related protein expressions were also regulated in SLT-treated cells. Moreover, DCFH-DA and western blot assays demonstrated that SLT enhanced ROS accumulation and subsequently activated the p53 signaling pathway. However, SLT-regulated ovarian cancer cell proliferation, apoptosis, migration, invasion, and EMT were significantly reversed by an ROS inhibitor (NAC, N-acetyl-L-cysteine). Furthermore, A2780 and SKOV3 cells cocultured with M0 macrophages showed that SLT activated the polarization of M0 macrophages to M1 macrophages and inhibited the polarization to M2 macrophages, with the increased percentage of CD86+ cells and decreased percentage of CD206+ cells were detected. In summary, this study illustrated the anticancer effects of SLT on ovarian cancer cells, suggesting that SLT may have the potential to provide basic evidence for the discovery of antiovarian cancer agents.

Lidocaine Protects Endometriosis by Inducing Apoptosis of Endometrial Stromal Cells

2020 ◽  
Vol 10 (6) ◽  
pp. 845-851
Author(s):  
Ping Li ◽  
Qian Zhang ◽  
Tao Li ◽  
Haibo Wang ◽  
Xia Ji ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Stromal Cells ◽  
Blot Analysis ◽  
Protein Level ◽  
Cell Apoptosis ◽  
Western Blot Analysis ◽  
Control Group ◽  
Endometrial Stromal Cells ◽  
Qrt Pcr

Objective: This study aimed to explore the effects of lidocaine on ectopic endometrial stromal cells and the underlying mechanisms in endometriosis. Methods: Ectopic endometrial stromal cells (ESCs), which were separated from endometriotic tissues, were subjected to various concentrations of lidocaine for different time. After the treatment of lidocaine, MTT assay was applied to assess the cell proliferation of ESCs, and cell apoptosis was analyzed by flow cytometry. Meanwhile, we detected the protein level of pro-caspse3 and cleaved-caspase3 protein in ESCs by Western blot analysis. The invasion and migration capability of ESCs was also detected by transwell assay. Western blot analysis and qRT-PCR were performed for the detection of EMT markers (E-cadherin, N-cadherin and MMP-9). Similarly, the expression levels of β-catenin, cyclin D1 and c-myc in ESCs were determined by Western blot analysis and qRT-PCR. Results: 0.5, 1, 5 and 10 mM of lidocaine obviously inhibited the cell proliferation of ESCs at different time points compared with the control group, while no significant effect was observed in 0.1 mM lidocaine treated cells. Lidocaine dose-dependently increased cell apoptosis, reduced the protein level of pro-caspse3 protein, but improved cleaved-caspase3 protein level in ESCs. Moreover, lidocaine dose-dependently decreased the migration and invasion capabilities of ESCs. In addition, compared to the control group, lidocaine enhanced the level of E-cadherin, and reduced the level of N-cadherin and MMP-9 in ESCs. Lidocaine suppressed the level of β-catenin, cyclin D1 and c-myc in ESCs in a dose-dependent manner. Conclusion: We demonstrated that lidocaine protected endometriosis by inducing the apoptosis of ESCs via regulating Wnt/β-catenin pathway.

scholarly journals Prognostic Impact of CEACAM1 in Node-Negative Ovarian Cancer Patients

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Leticia Oliveira-Ferrer ◽  
Roshni Goswami ◽  
Vladimir Galatenko ◽  
Yi Ding ◽  
Kathrin Eylmann ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Protein Expression ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Prognostic Impact ◽  
Cancer Type ◽  
Mrna Levels ◽  
Node Negative ◽  
Prognostic Relevance

The underlying mechanisms of ovarian cancer (OvCa) dissemination are still poorly understood, and novel molecular markers for this cancer type are urgently needed. In search of adhesion molecules with prognostic relevance in OvCa, we compared tumors with good outcome (alive > 3 years) and those with poor outcome (dead < 2 years) within data from The Cancer Genome Atlas (TCGA). The carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) turned out as the only gene with differential expression in these groups. In order to further investigation on its role in OvCa, we analyzed CEACAM1 mRNA levels extracted from TCGA microarray data (n=517) as well as CEACAM1 protein expression by Western blot analysis in a cohort of 242 tumor samples. Further, CEACAM1 localization in tumour tissue was evaluated by immunohistochemistry and CEACAM1 splice variants by RT-PCR in representative tumours. In Kaplan–Meier analysis, high CEACAM1 mRNA levels significantly correlated with longer survival (p=0.008). By Western blot analysis in the second cohort, similar associations of high CEACAM1 protein levels with longer recurrence-free survival (RFS, p=0.035) and overall survival (OAS, p=0.004) were observed. In multivariate Cox regression analysis including clinical prognostic parameters, CEACAM1 mRNA or protein expression turned out as independent prognostic markers. Stratified survival analysis showed that high CEACAM1 protein expression was prognostic in node-negative tumors (p=0.045 and p=0.0002 for DFS and OAS) but lost prognostic significance in node-positive carcinomas. Similarly, high CEACAM1 mRNA expression did not show prognostic relevance in tumors with lymphatic invasion (L1) but was associated with longer survival in cases without lymphovascular involvement. Further analysis showed a predominance of 4S and 4L isoforms and mostly membraneous CEACAM1 localization in ovarian tumours. Our results suggest that CEACAM1 might be an independent favorable prognostic marker in OvCa, especially in the subgroup of patients with solely intraperitoneal metastasis.

Synthesis and Characterization of Quinoline-3-Carboxamide Derivatives as Inhibitors of the ATM Kinase

2020 ◽  
Vol 20 (23) ◽  
pp. 2070-2079
Author(s):  
Srimadhavi Ravi ◽  
Sugata Barui ◽  
Sivapriya Kirubakaran ◽  
Parul Duhan ◽  
Kaushik Bhowmik
Keyword(s):  
Western Blot ◽  
Cancer Cells ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Cancer Cell Lines ◽  
Atm Kinase ◽  
Cytotoxicity Studies

Background: The importance of inhibiting the kinases of the DDR pathway for radiosensitizing cancer cells is well established. Cancer cells exploit these kinases for their survival, which leads to the development of resistance towards DNA damaging therapeutics. Objective: In this article, the focus is on targeting the key mediator of the DDR pathway, the ATM kinase. A new set of quinoline-3-carboxamides, as potential inhibitors of ATM, is reported. Methods: Quinoline-3-carboxamide derivatives were synthesized and cytotoxicity assay was performed to analyze the effect of molecules on different cancer cell lines like HCT116, MDA-MB-468, and MDA-MB-231. Results: Three of the synthesized compounds showed promising cytotoxicity towards a selected set of cancer cell lines. Western Blot analysis was also performed by pre-treating the cells with quercetin, a known ATM upregulator, by causing DNA double-strand breaks. SAR studies suggested the importance of the electron-donating nature of the R group for the molecule to be toxic. Finally, Western-Blot analysis confirmed the down-regulation of ATM in the cells. Additionally, the PTEN negative cell line, MDA-MB-468, was more sensitive towards the compounds in comparison with the PTEN positive cell line, MDA-MB-231. Cytotoxicity studies against 293T cells showed that the compounds were at least three times less toxic when compared with HCT116. Conclusion: In conclusion, these experiments will lay the groundwork for the evolution of potent and selective ATM inhibitors for the radio- and chemo-sensitization of cancer cells.

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