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We have not received reimbursements, fees, funding, or salary from an organization that may in any way gain or lose financially from the publication of this manuscript, either now or in the future. No such organization is financing this manuscript, including the article-processing charge. We do not hold stocks or shares in any organization that may gain or lose financially from the publication of this manuscript, either now or in the future. We do not hold and are not currently applying for any patents relating to the content of the manuscript.

We have not received reimbursements, fees, funding, or salary from an organization that holds or has applied for patents relating to the content of the manuscript. We have no other financial competing interests. We have no non-financial competing interests political, personal, religious, ideological, academic, intellectual, or commercial to declare in relation to this manuscript.

The author s declare that they have no competing interests. SRA generated the resistant cell lines, assisted in performing the microarray experiments, and performed the data subset analysis and selection of the validation genes. RN performed and analyzed the qPCR validation experiments for the selected genes. BG carried out the microarray experiments and the initial data analysis to identify genes with significantly different expression in the resistant cell lines.

AP contributed to the conception of the study, reviewed the manuscript and oversaw the microarray experiments. KLM performed the proliferation analysis of the cell lines and analyzed the data. SC carried out the immunoblots and generated the graphs. CL contributed to the conception of the study, oversaw the selection of the cell lines, oversaw the proliferation analysis, contributed to the final data analysis and drafted the manuscript. All authors read and approved the final manuscript.

Table S2. Determination of significant difference between the parent and resistant cell lines according to QPCR analysis. Figure S1. Comparison of gene expression changes between the resistant cell lines. The log fold change in gene expression is shown for each resistant cell line.

We thank the Genetics Laboratory at the Sudbury Regional Hospital for the use of the microarray scanner. National Center for Biotechnology Information , U. Journal List J Ovarian Res v. J Ovarian Res. Published online Nov Stephen R Armstrong 1 Dept. Rashmi Narendrula 1 Dept.

Amadeo M Parissenti 1 Dept. Author information Article notes Copyright and License information Disclaimer. Corresponding author. Stephen R Armstrong: ac. Received Jul 30; Accepted Oct This article has been cited by other articles in PMC. Primer sequences and melting temperatures. Additional file 2 Table S2.

Additional file 3 Figure S1. Abstract Background Current protocols for the treatment of ovarian cancer include combination chemotherapy with a platinating agent and a taxane. Methods The A epithelial endometrioid ovarian cancer cell line was used to select for isogenic carboplatin, docetaxel and dual drug resistant cell lines. Results Three isogenic cell lines were developed and resistance to each drug or the combination of drugs was confirmed.

Conclusions Ovarian tumor cells can acquire resistance to both carboplatin and docetaxel when selected in the presence of both agents. Determination of IC 50 The number of colonies growing was recorded by taking photomicrographs of five random fields X magnification per drug concentration and counting the colonies in each field.

Cell line selection Cell line selection was performed as described in a previous study by Guo et al. Cell line growth rate analysis Cells were plated at a density of 2. Statistical analysis of changes in gene expression For the microarray data, significant differences in fold change expression between co-cultured controls and resistant lines was determined using the Partek Genomics Suite program Partek Inc.

Table 1 Resistance values determined by clonogenic assay expressed as IC 50 for each cell line and drug s. Open in a separate window. Figure 1. Figure 2. Figure 3. Figure 4. Figure 5. Figure 6. Figure 7. Selection of validation gene sets Unique gene signals based on the microarray results were selected from each of the three resistant cell lines for validation by QPCR and immunoblotting.

Table 2 Validation gene set , genes with significant changes in one or more of the resistant cell lines according to microarray analysis. Table 3 Validation gene set , genes present in all three cell lines according to microarray analysis.

Significant differences confirmed between parent and resistant cell lines Significant differences in transcript levels from the microarray data were confirmed by QPCR for all transcripts in at least one cell line except the GSTO1 transcript, which was not found to be significantly different from the parent in any of the cell lines, despite the microarray results Additional file 2 : Table S2. The dual resistant line contains specific differences in gene expression To examine if the expression level of the selected genes was significantly different between the resistant cell lines, one-way ANOVA was performed on the log of the fold change as determined by QPCR.

Changes in protein expression determined by immunoblotting Further confirmation of the changes in gene expression, at the protein level, was attempted by immunoblotting. Figure 8. Figure 9. Discussion Selection of resistant cell lines In this study, a set of three isogenic drug-resistant ovarian cancer cell lines has been generated from the A ovarian cancer cell line.

Characterization of levels of resistance During the selection for single or dual drug resistance in our study, the gradual increase in drug concentration, beginning with a dose fold below the IC 50 of the parental A cell line, generated populations of resistant cells and avoided selection of a few drug resistant clones.

Immunoblot confirmation of changes in protein expression Immunoblots were performed to determine if changes in gene expression at the transcript level could be confirmed at the protein expression level. Conclusions In this study, we report the establishment of a novel cell line with documented resistance to both carboplatin and docetaxel.

Competing interests The authors of this paper have no competing interests to declare. Supplementary Material Additional file 1: Table S1. Click here for file 47K, doc. Additional file 2: Table S2. Click here for file 44K, doc.

Additional file 3: Figure S1. Click here for file K, docx. Acknowledgements We thank the Genetics Laboratory at the Sudbury Regional Hospital for the use of the microarray scanner. Cancer Statistics, CA Cancer J Clin. Intraperitoneal chemotherapy in the first-line treatment of women with stage III epithelial ovarian cancer: a systematic review with metaanalyses. Antineoplastic agents in the management of ovarian cancer: current status and emerging therapeutic strategies.

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Combined oral cyclophosphamide and bevacizumab in heavily pre-treated ovarian cancer. Clin Transl Oncol. Carboplatin and paclitaxel versus cisplatin, paclitaxel and doxorubicin for first-line chemotherapy of advanced ovarian cancer: a Hellenic Cooperative Oncology Group HeCOG study.

Eur J Cancer. Phase III trial of carboplatin and paclitaxel compared with cisplatin and paclitaxel in patients with optimally resected stage III ovarian cancer: a Gynecologic Oncology Group study. Eur J Cancer Care Engl ; 19 2 — Cross-resistance studies of isogenic drug-resistant breast tumor cell lines support recent clinical evidence suggesting that sensitivity to paclitaxel may be strongly compromised by prior doxorubicin exposure.

Potent killing of paclitaxel- and doxorubicin-resistant breast cancer cells by calphostin C accompanied by cytoplasmic vacuolization. Overexpression of dihydrodiol dehydrogenase is associated with cisplatin-based chemotherapy resistance in ovarian cancer patients.

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BMC Med Genomics. Increased DNA repair as a mechanism of acquired resistance to cis-diamminedichloroplatinum II in human ovarian cancer cell lines. Targeting tumor-associated fibroblasts improves cancer chemotherapy by increasing intratumoral drug uptake. J Clin Invest.

Antiangiogenesis enhances intratumoral drug retention. Drug resistance and the solid tumor microenvironment. J Cancer Res Clin Oncol. Phase I trial of intraperitoneal docetaxel in the treatment of advanced malignancies primarily confined to the peritoneal cavity: dose-limiting toxicity and pharmacokinetics. Description of paclitaxel resistance-associated genes in ovarian and breast cancer cell lines. Molecular description of evolving paclitaxel resistance in the SKOV-3 human ovarian carcinoma cell line.

In vitro cross-resistance and collateral sensitivity in seven resistant small-cell lung cancer cell lines: preclinical identification of suitable drug partners to taxotere, taxol, topotecan and gemcitabin. Mitomycin C cross-resistance induced by adriamycin in human ovarian cancer cells in vitro.

Platinum-Taxol non-cross resistance in epithelial ovarian cancer. Peritoneal mesothelial hyperplasia associated with gynaecological disease: a potential diagnostic pitfall that is commonly associated with endometriosis. J Clin Pathol. The putative cannabinoid receptor GPR55 defines a novel autocrine loop in cancer cell proliferation. Nucl Med Biol. Nrf2 enhances cell proliferation and resistance to anticancer drugs in human lung cancer.

Cell proliferation and drug resistance in hepatocellular carcinoma are modulated by Rho GTPase signals. Cell proliferation is related to in vitro drug resistance in childhood acute leukaemia. Cell adhesion is a key determinant in de novo multidrug resistance MDR : new targets for the prevention of acquired MDR.

An integrated database of chemosensitivity to 55 anticancer drugs and gene expression profiles of 39 human cancer cell lines. Increased expression of dihydrodiol dehydrogenase induces resistance to cisplatin in human ovarian carcinoma cells. J Biol Chem. Olaparib in patients with recurrent high-grade serous or poorly differentiated ovarian carcinoma or triple-negative breast cancer: a phase 2, multicentre, open-label, non-randomised study.

Lancet Oncol. Tumor growth inhibition by olaparib in BRCA2 germline-mutated patient-derived ovarian cancer tissue xenografts. Making the best of PARP inhibitors in ovarian cancer. Nat Rev Clin Oncol. Sci Signal. Otolaryngol Head Neck Surg.

Analysis of epithelial and mesenchymal markers in ovarian cancer reveals phenotypic heterogeneity and plasticity. PLoS One. MUC16 CA regulates epithelial ovarian cancer cell growth, tumorigenesis and metastasis. Antiadhesive antibodies targeting E-cadherin sensitize multicellular tumor spheroids to chemotherapy in vitro.

N-cadherin and cadherin 11 modulate postnatal bone growth and osteoblast differentiation by distinct mechanisms. J Cell Sci. The intracellular domain of cadherin is not required for the induction of cell aggregation, adhesion or gap-junction formation. Cell Commun Adhes. Building the synovium: cadherin mediates fibroblast-like synoviocyte cell-to-cell adhesion.

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