The Profile of MicroRNA Expression and Potential Role in the Regulation of Drug-Resistant Genes in Cisplatin- and Paclitaxel-Resistant Ovarian Cancer Cell Lines

Ovarian cancer is the most lethal gynecological malignancy. The high mortality results from late diagnosis and the development of drug resistance. Drug resistance results from changes in the expression of different drug-resistance genes that may be regulated miRNA. The main aim of our study was to detect changes in miRNA expression levels in two cisplatin (CIS) and two paclitaxel (PAC)—resistant variants of the A2780 drug-sensitive ovarian cancer cell line—by miRNA microarray. The next goal was to identify miRNAs responsible for the regulation of drug-resistance genes. We observed changes in the expression of 46 miRNA that may be related to drug resistance. The overexpression of miR-125b-5p, miR-99a-5p, miR-296-3p, and miR-887-3p and downregulation of miR-218-5p, miR-221-3p, and miR-222-3p was observed in both CIS-resistant cell lines. In both PAC-resistant cell lines, we observed the upregulation of miR-221-3p, miR-222-3p, and miR-4485, and decreased expression of miR-551b-3p, miR-551b-5p, and miR-218-5p. Analysis of targets suggest that expression of important drug-resistant genes like protein Tyrosine Phosphatase Receptor Type K (PTPRK), receptor tyrosine kinase—EPHA7, Semaphorin 3A (SEMA3A), or the ATP-binding cassette subfamily B member 1 gene (ABCB1) can be regulated by miRNA.

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Expression of Osteoblast-Specific Factor 2 (OSF-2, Periostin) Is Associated with Drug Resistance in Ovarian Cancer Cell Lines

International Journal of Molecular Sciences ◽

10.3390/ijms20163927 ◽

2019 ◽

Vol 20 (16) ◽

pp. 3927 ◽

Cited By ~ 3

Author(s):

Karolina Sterzyńska ◽

Dominika Kaźmierczak ◽

Andrzej Klejewski ◽

Monika Świerczewska ◽

Karolina Wojtowicz ◽

...

Keyword(s):

Ovarian Cancer ◽

Drug Resistance ◽

Cell Line ◽

Cell Lines ◽

Cancer Cell ◽

Ovarian Cancer Cell ◽

Ovarian Cancer Cell Line ◽

Cancer Cell Line ◽

Specific Factor ◽

Drug Resistant

One of the main obstacles to the effective treatment of ovarian cancer patients continues to be the drug resistance of cancer cells. Osteoblast-Specific Factor 2 (OSF-2, Periostin) is a secreted extracellular matrix protein (ECM) expressed in fibroblasts during bone and teeth development. Expression of OSF-2 has been also related to the progression and drug resistance of different tumors. The present study investigated the role of OSF-2 by evaluating its expression in the primary serous ovarian cancer cell line, sensitive (W1) and resistant to doxorubicin (DOX) (W1DR) and methotrexate (MTX) (W1MR). The OSF-2 transcript (real-time PCR analysis), protein expression in cell lysates and cell culture medium (western blot), and expression of the OSF-2 protein in cell lines (immunofluorescence) were investigated in this study. Increased expression of OSF-2 mRNA was observed in drug-resistant cells and followed by increased protein expression in cell culture media of drug-resistant cell lines. A subpopulation of ALDH1A1-positive cells was noted for W1DR and W1MR cell lines; however, no direct co-expression with OSF-2 was demonstrated. Both drugs induced OSF-2 expression after a short period of exposure of the drug-sensitive cell line to DOX and MTX. The obtained results indicate that OSF-2 expression might be associated with the development of DOX and MTX resistance in the primary serous W1 ovarian cancer cell line.

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The Significance of MicroRNAs Expression in Regulation of Extracellular Matrix and Other Drug Resistant Genes in Drug Resistant Ovarian Cancer Cell Lines

International Journal of Molecular Sciences ◽

10.3390/ijms21072619 ◽

2020 ◽

Vol 21 (7) ◽

pp. 2619 ◽

Cited By ~ 1

Author(s):

Dominika Kazmierczak ◽

Karol Jopek ◽

Karolina Sterzynska ◽

Barbara Ginter-Matuszewska ◽

Michal Nowicki ◽

...

Keyword(s):

Ovarian Cancer ◽

Extracellular Matrix ◽

Drug Resistance ◽

Cell Line ◽

Cancer Cell ◽

Ovarian Cancer Cell ◽

Resistant Cell ◽

Resistant Cell Line ◽

Drug Resistant ◽

Resistant Genes

Ovarian cancer rates the highest mortality among all gynecological malignancies. The main reason for high mortality is the development of drug resistance. It can be related to increased expression of drug transporters and increased expression of extracellular matrix (ECM) proteins. Our foremost aim was to exhibit alterations in the miRNA expression levels in cisplatin (CIS), paclitaxel (PAC), doxorubicin (DOX), and topotecan (TOP)—resistant variants of the W1 sensitive ovarian cancer cell line—using miRNA microarray. The second goal was to identify miRNAs responsible for the regulation of drug-resistant genes. According to our observation, alterations in the expression of 40 miRNAs were present. We could observe that, in at least one drug-resistant cell line, the expression of 21 miRNAs was upregulated and that of 19 miRNAs was downregulated. We identified target genes for 22 miRNAs. Target analysis showed that miRNA regulates key genes responsible for drug resistance. Among others, we observed regulation of the ATP-binding cassette subfamily B member 1 gene (ABCB1) in the paclitaxel-resistant cell line by miR-363 and regulation of the collagen type III alpha 1 chain gene (COL3A1) in the topotekan-resistant cell line by miR-29a.

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MDR Gene Expression Analysis of Six Drug-Resistant Ovarian Cancer Cell Lines

BioMed Research International ◽

10.1155/2013/241763 ◽

2013 ◽

Vol 2013 ◽

pp. 1-10 ◽

Cited By ~ 55

Author(s):

Radosław Januchowski ◽

Karolina Wojtowicz ◽

Patrycja Sujka-Kordowska ◽

Małgorzata Andrzejewska ◽

Maciej Zabel

Keyword(s):

Ovarian Cancer ◽

Drug Resistance ◽

Cell Lines ◽

Cancer Cell ◽

Ovarian Cancer Cell ◽

Multiple Drug Resistance ◽

Cancer Cell Lines ◽

Resistant Cell ◽

Cross Resistance ◽

Ovarian Cancer Cell Lines

Ovarian cancer is the leading cause of death among gynaecological malignancies. Multiple drug resistance makes cancer cells insensitive to chemotherapy. In this study, we developed six primary ovarian cancer cell lines (W1MR, W1CR, W1DR, W1VR, W1TR, and W1PR) resistant to drugs such as methotrexate, cisplatin, doxorubicin, vincristine, topotecan, and paclitaxel. A chemosensitivity assay MTT test was performed to assess drug cross-resistance. Quantitative real-time polymerase chain reaction and Western blot were also performed to determine mRNA and protein expression of genes involved in chemoresistance. We observed high cross-resistance to doxorubicin, vincristine, and paclitaxel in the cell lines resistant to these agents. We also found a significant correlation between resistance to these drugs and increased expression of P-gp. Two different mechanisms of topotecan resistance were observed in the W1TR and W1PR cell lines. We did not observe any correlation between MRP2 transcript and protein levels. Cell lines resistant to agents used in ovarian cancer treatment remained sensitive to methotrexate. The main mechanisms of drug resistance were due to P-gp expression in the doxorubicin, vincristine, and paclitaxel resistant cell lines and BCRP expression in the topotecan resistant cell line.

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The significance of microRNA genes expression in ovarian cancer cell lines resistant to cytotoxic drugs

10.1101/859264 ◽

2019 ◽

Author(s):

Dominika Kazmierczak ◽

Karol Jopek ◽

Karolina Sterzynska ◽

Marcin Rucinski ◽

Radoslaw Januchowski

Keyword(s):

Ovarian Cancer ◽

Drug Resistance ◽

Cell Line ◽

Cancer Cell ◽

Ovarian Cancer Cell ◽

Gynecological Cancer ◽

Resistant Cell ◽

Resistant Cell Line ◽

Drug Resistant ◽

Drug Resistant Cell

ABSTRACTOvarian cancer is the most fatal type of gynecological cancer. The main reason for high mortality is the development of drug resistance. It can be related to increased expression of drug transporters and increased expression of extracellular matrix (ECM) proteins. miRNA is a short noncoding RNA that regulates expression of about 60% of genes in the human genome and plays a crucial role in developing cancer, including resistance to chemotherapy.Our foremost aim was to exhibit alterations in the miRNA expression levels in the cisplatin (CIS), paclitaxel (PAC), doxorubicin (DOX), and topotecan (TOP) - resistant variants of W1 sensitive ovarian cancer cell line using miRNA microarray. The second goal was to identify miRNAs responsible for the regulation of drug-resistant genes.According to our observation, alterations in the expression of 40 miRNA genes. The level of expression of 21 genes was upregulated in at least one drug-resistant cell line. The expression of 19 genes was downregulated in at least one drug-resistant cell line. We identified target genes for 22 miRNAs. Target analysis showed that miRNA regulates key genes responsible for drug resistance. Among others, we observed regulation of the ABCB1 (MDR1) gene in the W1PR1 cell line by miR-363 and regulation of the COL3A1 gene in the W1TR cell line by miR-29a.Hence, on the basis of our findings, results indicate that genes responsible for drug resistance development in ovarian cancer can be regulated by miRNA.

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Extracellular Matrix Proteins Expression Profiling in Chemoresistant Variants of the A2780 Ovarian Cancer Cell Line

BioMed Research International ◽

10.1155/2014/365867 ◽

2014 ◽

Vol 2014 ◽

pp. 1-9 ◽

Cited By ~ 38

Author(s):

Radosław Januchowski ◽

Piotr Zawierucha ◽

Marcin Ruciński ◽

Michał Nowicki ◽

Maciej Zabel

Keyword(s):

Ovarian Cancer ◽

Extracellular Matrix ◽

Drug Resistance ◽

Cell Line ◽

Cancer Cell ◽

Ovarian Cancer Cell ◽

Ovarian Cancer Cell Line ◽

Cancer Cell Line ◽

Altered Expression ◽

Expression Levels

Ovarian cancer is the leading cause of death among gynaecological malignancies. Extracellular matrix (ECM) can affect drug resistance by preventing the penetration of the drug into cancer cells and increased resistance to apoptosis. This study demonstrates alterations in the expression levels of ECM components and related genes in cisplatin-, doxorubicin-, topotecan-, and paclitaxel-resistant variants of the A2780 ovarian cancer cell line. Affymetrix Gene Chip Human Genome Array Strips were used for hybridisations. The genes that had altered expression levels in drug-resistant sublines were selected and filtered by scatter plots. The genes that were up- or downregulated more than fivefold were selected and listed. Among the investigated genes, 28 genes were upregulated, 10 genes were downregulated, and two genes were down- or upregulated depending on the cell line. Between upregulated genes 12 were upregulated very significantly—over 20-fold. These genes included COL1A2, COL12A1, COL21A1, LOX, TGFBI, LAMB1, EFEMP1, GPC3, SDC2, MGP, MMP3, and TIMP3. Four genes were very significantly downregulated: COL11A1, LAMA2, GPC6, and LUM. The expression profiles of investigated genes provide a preliminary insight into the relationship between drug resistance and the expression of ECM components. Identifying correlations between investigated genes and drug resistance will require further analysis.

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Microarray-based detection and expression analysis of new genes associated with drug resistance in ovarian cancer cell lines

Oncotarget ◽

10.18632/oncotarget.18278 ◽

2017 ◽

Vol 8 (30) ◽

pp. 49944-49958 ◽

Cited By ~ 28

Author(s):

Radosław Januchowski ◽

Karolina Sterzyńska ◽

Piotr Zawierucha ◽

Marcin Ruciński ◽

Monika Świerczewska ◽

...

Keyword(s):

Ovarian Cancer ◽

Drug Resistance ◽

Cell Lines ◽

Cancer Cell ◽

Expression Analysis ◽

Ovarian Cancer Cell ◽

Cancer Cell Lines ◽

New Genes ◽

Ovarian Cancer Cell Lines

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Inhibition of MUS81 By siRNA Reverses Resistance to Cisplatin in Human Ovarian Cancer Cell Lines

10.21203/rs.3.rs-667572/v2 ◽

2021 ◽

Author(s):

Emma C. Bourton ◽

Sheba Adam-Zahir ◽

Piers N. Plowman ◽

Hussein Nahidh Al-Ali ◽

Helen A. Foster ◽

...

Keyword(s):

Ovarian Cancer ◽

Protein Expression ◽

Cell Lines ◽

Cancer Cell ◽

Ovarian Cancer Cell ◽

Cancer Cell Lines ◽

Resistant Cell ◽

Wild Type ◽

Ovarian Cancer Cell Lines ◽

Sirna Knockdown

Abstract Bacground: Drugs that induce DNA interstrand crosslinks form the mainstay of anticancer treatments for different cancers. These drugs are used to treat ovarian cancer which is the most prevalent gynaecological cancer. Five-year survival rates are approximately 40% and the development of drug resistant disease is an important factor in treatment failure. Methods: In this study a comprehensive evaluation of the expression and function of the site-specific endonuclease MUS81 was conducted. Using quantitative real time PCR analysis and imaging flow cytometry we determined the mRNA and protein expression of MUS81 in three ovarian cancer cell lines and two immortalised human fibroblast cell lines which had been made resistant to cisplatin by chronic exposure. siRNA knockdown of MUS81 was employed to determine the effect on overall cell survival which was assessed using clonogenic assays. Results: In the five cisplatin-resistant cell lines we observed increased MUS81 mRNA expression. In addition MUS81 protein expression in the form of discrete nuclear foci in cells was observed in all cell lines following cisplatin exposure, there being significantly more foci in cisplatin resistant cell lines. siRNA knockdown of MUS81 significantly reduced both mRNA and protein levels in two cell lines (SK-OV-3 and MRC5-SV1 – wild-type and resistant) and critically re-sensitised cisplatin resistant cells to wild-type level, determined by clonogenic assay.Conclusion: MUS81 is central to the development of cisplatin resistance in ovarian cancer cell lines. Inhibition of MUS81 restored drug sensitivity to the cells. MUS81 may be a useful therapeutic target to overcome drug resistance in ovarian and other cancers.

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Coexisting Molecular Determinants of Acquired Oxaliplatin Resistance in Human Colorectal and Ovarian Cancer Cell Lines

International Journal of Molecular Sciences ◽

10.3390/ijms20153619 ◽

2019 ◽

Vol 20 (15) ◽

pp. 3619 ◽

Cited By ~ 3

Author(s):

Paul Noordhuis ◽

Adrianus C. Laan ◽

Kasper van de Born ◽

Richard J. Honeywell ◽

Godefridus J. Peters

Keyword(s):

Gene Expression ◽

Ovarian Cancer ◽

Cell Lines ◽

Cancer Cell ◽

Array Cgh ◽

Ovarian Cancer Cell ◽

Ovarian Cancer Cell Line ◽

Continuous Exposure ◽

Platinum Accumulation ◽

Repair Genes

Oxaliplatin (OHP) treatment of colorectal cancer (CRC) frequently leads to resistance. OHP resistance was induced in CRC cell lines LoVo-92 and LoVo-Li and a platinum-sensitive ovarian cancer cell line, A2780, and related to cellular platinum accumulation, platinum-DNA adducts, transporter expression, DNA repair genes, gene expression arrays, and array-CGH profiling. Pulse (4 h, 4OHP) and continuous exposure (72 h, cOHP) resulted in 4.0 to 7.9-fold and 5.0 to 11.8-fold drug resistance, respectively. Cellular oxaliplatin accumulation and DNA-adduct formation were decreased and related to OCT1-3 and ATP7A expression. Gene expression profiling and pathway analysis showed significantly altered p53 signaling, xenobiotic metabolism, role of BRCA1 in DNA damage response, and aryl hydrocarbon receptor signaling pathways, were related to decreased ALDH1L2, Bax, and BBC3 (PUMA) and increased aldo-keto reductases C1 and C3. The array-CGH profiles showed focal aberrations. In conclusion, OHP resistance was correlated with total platinum accumulation and OCT1-3 expression, decreased proapoptotic, and increased anti-apoptosis and homologous repair genes.

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Mutual Expression of ALDH1A1, LOX, and Collagens in Ovarian Cancer Cell Lines as Combined CSCs- and ECM-Related Models of Drug Resistance Development

International Journal of Molecular Sciences ◽

10.3390/ijms20010054 ◽

2018 ◽

Vol 20 (1) ◽

pp. 54 ◽

Cited By ~ 9

Author(s):

Karolina Sterzyńska ◽

Andrzej Klejewski ◽

Karolina Wojtowicz ◽

Monika Świerczewska ◽

Marta Nowacka ◽

...

Keyword(s):

Ovarian Cancer ◽

Extracellular Matrix ◽

Drug Resistance ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Lysyl Oxidase ◽

Resistant Cell ◽

Pcr Analysis ◽

Ovarian Cancer Cells

A major contributor leading to treatment failure of ovarian cancer patients is the drug resistance of cancer cell. CSCs- (cancer stem cells) and ECM (extracellular matrix)-related models of drug resistance are described as independently occurring in cancer cells. Lysyl oxidase (LOX) is another extracellular protein involved in collagen cross-linking and remodeling of extracellular matrix and has been correlated with tumor progression. The expression of LOX, COL1A2, COL3A1, and ALDH1A1 was performed in sensitive (A2780, W1) and resistant to paclitaxel (PAC) (A2780PR1 and W1PR2) and topotecan (TOP) (W1TR) cell lines at the mRNA (real-time PCR analysis) and protein level (Western blot and immunofluorescence analysis). The ALDH1A1 activity was measured with the ALDEFLUOR test and flow cytometry analysis. The protein expression in ovarian cancer tissues was determined by immunohistochemistry. We observed an increased expression of LOX and collagens in PAC and TOP resistant cell lines. Subpopulations of ALDH1A1 positive and negative cells were also noted for examined cell lines. Additionally, the coexpression of LOX with ALDH1A1 and COL1A2 with ALDH1A1 was observed. The expression of LOX, collagens, and ALDH1A1 was also detected in ovarian cancer lesions. In our study LOX, ALDH1A1 and collagens were found to be coordinately expressed by cells resistant to PAC (LOX, ALDH1A1, and COL1A2) or to TOP (LOX and ALDH1A1). This represents the study where molecules related with CSCs (ALDH1A1) and ECM (LOX, collagens) models of drug resistance are described as occurring simultaneously in ovarian cancer cells treated with PAC and TOP.

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