Drug transporter expression profiling in chemoresistant variants of the A2780 ovarian cancer cell line.

Ovarian cancer is characterized by the higher mortality among gynecological cancers. In results of MDR development during chemotherapy cancer cells become resistant to further treatment. Microarray techniques can provide information about MDR development at gene expression level. ABC and SLC transporters are most important proteins responsible for this phenomenon. In this study changes of ABC and SLC genes expression pattern in drugs resistant sublines of the A2780 ovarian cancer cell line were demonstrated. The cytostatic resistant sublines were generated by culture of A2780 cell line with an increasing concentration of the indicated drugs. As screening methods, we used Affymetrix U219 Human Genome microarrays. Independent t-tests were used to determinate statistical significances of results. Genes that expression levels were higher than assumed threshold (upregulated above threefold and downregulated under -3 fold) were visualized using scatter plot method, selected and listed in table. Between the ABC genes increased expression of seven genes and decreased expression of three genes were observed. Expression of two genes was increased or decreased depending on the cell line. We observed significant (more than tenfold) increase in expression of four ABC genes: ABCA8, ABCB1, ABCB4 and ABCG2 and decreased expression of ABCA3 gene. We also observed changes in expression of 32 SLC genes. Between them we observe increased expression of 17 genes and decreased expression of 15 genes. Expression of four genes was increased or decreased dependent on cell line. The expression of nine SLC genes increased or decreased very significantly (more than tenfold). Five genes were significantly upregulated: SLC2A9, SLC16A3, SLC16A14, SLC38A4 and SLC39A8. Four additional genes were significantly downregulated: SLC2A14, SLC6A15, SLC8A1 and SLC27A2. Expression profiles of these genes give strong arguments for assumption of correlation between expression of ABC and SLC genes and drug resistance phenomenon. Identifying correlations between specific drug transporters and cytostatic drug resistance will require further investigation.

[1]  H. Yoshikawa,et al.  Role of cystine transport in intracellular glutathione level and cisplatin resistance in human ovarian cancer cell lines , 2003, British Journal of Cancer.

[2]  N. Cheong,et al.  Intracellular ABC transporter A3 confers multidrug resistance in leukemia cells by lysosomal drug sequestration , 2008, Leukemia.

[3]  Ying Huang,et al.  Membrane transporters and channels in chemoresistance and -sensitivity of tumor cells. , 2006, Cancer letters.

[4]  M. Pešić,et al.  The role of paclitaxel in the development and treatment of multidrug resistant cancer cell lines. , 2011, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[5]  F. de Longueville,et al.  ABCA3 as a Possible Cause of Drug Resistance in Childhood Acute Myeloid Leukemia , 2006, Clinical Cancer Research.

[6]  V. Torri,et al.  Paclitaxel plus platinum-based chemotherapy versus conventional platinum-based chemotherapy in women with relapsed ovarian cancer: the ICON4/AGO-OVAR-2.2 trial , 2003, The Lancet.

[7]  M. Stauch,et al.  Nonplatinum topotecan combinations versus topotecan alone for recurrent ovarian cancer: results of a phase III study of the North-Eastern German Society of Gynecological Oncology Ovarian Cancer Study Group. , 2008, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[8]  V. Gebski,et al.  A randomized, phase III study of carboplatin and pegylated liposomal doxorubicin versus carboplatin and paclitaxel in relapsed platinum-sensitive ovarian cancer (OC): CALYPSO study of the Gynecologic Cancer Intergroup (GCIG). , 2009, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[9]  T. Fojo,et al.  The role of ABC transporters in clinical practice. , 2003, The oncologist.

[10]  M. Dietel,et al.  In vitro prediction of cytostatic drug resistance in primary cell cultures of solid malignant tumours. , 1993, European journal of cancer.

[11]  S. Bates,et al.  ABCG2: determining its relevance in clinical drug resistance , 2007, Cancer and Metastasis Reviews.

[12]  Manfred Dietel,et al.  ABCC2 (MRP2, cMOAT) Can Be Localized in the Nuclear Membrane of Ovarian Carcinomas and Correlates with Resistance to Cisplatin and Clinical Outcome , 2006, Clinical Cancer Research.

[13]  P. Gros,et al.  Two members of the mouse mdr gene family confer multidrug resistance with overlapping but distinct drug specificities , 1990, Molecular and cellular biology.

[14]  T. Nakanishi.  Drug transporters as targets for cancer chemotherapy. , 2007, Cancer genomics & proteomics.

[15]  M. Burt,et al.  Rapid activation of MDR1 gene expression in human metastatic sarcoma after in vivo exposure to doxorubicin. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[16]  S. Byun,et al.  Augmentation of cisplatin sensitivity in cisplatin‐resistant human bladder cancer cells by modulating glutathione concentrations and glutathione‐related enzyme activities , 2005, BJU international.

[17]  Z. Duan,et al.  Description of paclitaxel resistance-associated genes in ovarian and breast cancer cell lines , 2005, Cancer Chemotherapy and Pharmacology.

[18]  D. Katsaros,et al.  Phase III trial of gemcitabine compared with pegylated liposomal doxorubicin in progressive or recurrent ovarian cancer. , 2008, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[19]  D. Schadendorf,et al.  Overexpression of cMOAT (MRP2/ABCC2) is associated with decreased formation of platinum-DNA adducts and decreased G2-arrest in melanoma cells resistant to cisplatin. , 2003, The Journal of investigative dermatology.

[20]  P. Sujka-Kordowska,et al.  MDR Gene Expression Analysis of Six Drug-Resistant Ovarian Cancer Cell Lines , 2012, BioMed research international.

[21]  A. Hall,et al.  Sensitivity of testis tumour cells to chemotherapeutic drugs: role of detoxifying pathways. , 1996, European journal of cancer.

[22]  F. Bolck.  Blaustein's Pathology of the Female Genital Tract, 3rd edition, R.J. Kurman (Ed.). Springer-Verlag, New York-Berlin-Heidelberg-London-Paris-Tokyo (1987), 959 pages, with 1176 figures in 1355 parts, 22 in full color, DM 298. (Distribution rights for Japan: Igaku Shoin, Tokyo), ISBN: 3-540-96452-5 , 1989 .

[23]  M. Gottesman,et al.  Multidrug resistance in cancer: role of ATP–dependent transporters , 2002, Nature Reviews Cancer.

[24]  P. Zawierucha,et al.  Microarray-based detection and expression analysis of ABC and SLC transporters in drug-resistant ovarian cancer cell lines. , 2013, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[25]  Z. Siddik,et al.  Cisplatin: mode of cytotoxic action and molecular basis of resistance , 2003, Oncogene.

[26]  T. Overbeck,et al.  Intracellular ATP-Binding Cassette Transporter A3 is Expressed in Lung Cancer Cells and Modulates Susceptibility to Cisplatin and Paclitaxel , 2013, Oncology.

[27]  R C Young,et al.  Characterization of a cis-diamminedichloroplatinum(II)-resistant human ovarian cancer cell line and its use in evaluation of platinum analogues. , 1987, Cancer research.

[28]  T. Ozben.  Mechanisms and strategies to overcome multiple drug resistance in cancer , 2006, FEBS letters.

[29]  J. Thigpen.  Gemcitabine Plus Carboplatin Compared With Carboplatin in Patients With Platinum-Sensitive Recurrent Ovarian Cancer: An Intergroup Trial of the AGO-OVAR, the NCIC CTG, and the EORTC GCG , 2008 .

[30]  J. Thigpen.  Randomized Phase III Trial of Gemcitabine Compared With Pegylated Liposomal Doxorubicin in Patients With Platinum-Resistant Ovarian Cancer , 2008 .

[31]  Ying Huang.  Pharmacogenetics/genomics of membrane transporters in cancer chemotherapy , 2007, Cancer and Metastasis Reviews.

[32]  F. Zunino,et al.  Ovarian cancer cisplatin-resistant cell lines: multiple changes including collateral sensitivity to Taxol. , 1998, Annals of oncology : official journal of the European Society for Medical Oncology.

[33]  Andreas Rolfs,et al.  The ABCs of solute carriers: physiological, pathological and therapeutic implications of human membrane transport proteins , 2004, Pflügers Archiv.

[34]  Y. Chiew,et al.  MRP2 (ABCC2) and cisplatin sensitivity in hepatocytes and human ovarian carcinoma. , 2006, Gynecologic oncology.

[35]  A. Fujimura,et al.  Functional analysis of ABCA8, a new drug transporter. , 2002, Biochemical and biophysical research communications.

[36]  R. Kurman,et al.  BLAUSTEIN'S PATHOLOGY OF THE FEMALE GENITAL TRACT , 2004 .

[37]  J. Schellens,et al.  Advances in Brief Overexpression of the BCRP / MXR / ABCP Gene in a Topotecan-selected Ovarian Tumor Cell Line 1 , 1999 .

[38]  C. Choi,et al.  ABC transporters as multidrug resistance mechanisms and the development of chemosensitizers for their reversal , 2005, Cancer Cell International.