Spectrin αII and βII tetramers contribute to platinum anticancer drug resistance in ovarian serous adenocarcinoma

To overcome drug resistance in ovarian carcinoma, novel resistance mechanisms must be elucidated for clinical application. We purified 2 proteins in the 300 kDa range from cisplatin‐resistant cells (NOS2CR2) by affinity chromatography with cisplatin‐exposed Glutathione Sepharose 4B. The purified proteins were identified as spectrin αII and βII by peptide mass mapping analysis. Western blot analysis detected greater expression of spectrin αII and βII in NOS2CR2 than in wild‐type cells (NOS2). The same result was obtained for spectrin βII expression by immunohistochemical staining. To determine whether spectrin αII and βII contribute to resistance, a drug sensitivity test was performed on SKOV3 ovarian cancer cells transfected with small interfering RNA. Sensitivity to platinum drugs was increased in the expression reduced cells. In a clinical study of five ovarian serous adenocarcinoma cases, tumor specimens taken after treatment with carboplatin stained more strongly for spectrin βII expression than untreated specimens. Fifty‐two tumor specimens from 46 patients with ovarian serous adenocarcinoma were immunohistochemically stained for spectrin βII and scored. Tumors previously treated by chemotherapy scored higher than those not treated. Of 27 cases with detectable residual tumors at the time of surgery, cases scoring 4–6 had shorter progression‐free survival periods after platinum‐based chemotherapy than cases scoring 0–3 (p = 0.012). The cytoskeleton proteins Spectrin αII and βII contributed to drug resistance by anchoring the GS‐Pt complex to the cell membrane, arresting cisplatin activity. Thus spectrin βII may be a useful predictor of platinum sensitivity in ovarian serous adenocarcinoma.

[1]  D. Gibson.  The mechanism of action of platinum anticancer agents--what do we really know about it? , 2009, Dalton transactions.

[2]  D. Gibson,et al.  Is glutathione the major cellular target of cisplatin? A study of the interactions of cisplatin with cancer cell extracts. , 2009, Journal of medicinal chemistry.

[3]  H. Kajiyama,et al.  Possible involvement of TWIST in enhanced peritoneal metastasis of epithelial ovarian carcinoma , 2007, Clinical & Experimental Metastasis.

[4]  S. Kyo,et al.  High Twist expression is involved in infiltrative endometrial cancer and affects patient survival. , 2006, Human pathology.

[5]  Dong Wang,et al.  Cellular processing of platinum anticancer drugs , 2005, Nature Reviews Drug Discovery.

[6]  T. Hasegawa,et al.  Effect of Antibiotics on Group A Streptococcus Exoprotein Production Analyzed by Two-Dimensional Gel Electrophoresis , 2005, Antimicrobial Agents and Chemotherapy.

[7]  J. Hilden,et al.  Should CA-125 response criteria be preferred to response evaluation criteria in solid tumors (RECIST) for prognostication during second-line chemotherapy of ovarian carcinoma? , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[8]  P. Bignone,et al.  Spectrin alpha II and beta II isoforms interact with high affinity at the tetramerization site. , 2003, The Biochemical journal.

[9]  T. Bauknecht,et al.  A randomized clinical trial of cisplatin/paclitaxel versus carboplatin/paclitaxel as first-line treatment of ovarian cancer. , 2003, Journal of the National Cancer Institute.

[10]  Robert S Mannel,et al.  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. , 2003, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[11]  I. Herskowitz,et al.  Uptake of the anticancer drug cisplatin mediated by the copper transporter Ctr1 in yeast and mammals , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[12]  G. Rustin,et al.  CA125 response: can it replace the traditional response criteria in ovarian cancer? , 2002, The oncologist.

[13]  J. Hayes,et al.  Glutathione S‐transferases , 2002 .

[14]  A. Baines,et al.  Spectrin and ankyrin-based pathways: metazoan inventions for integrating cells into tissues. , 2001, Physiological reviews.

[15]  E. Heerkens,et al.  Identification of a novel C-terminal variant of beta II spectrin: two isoforms of beta II spectrin have distinct intracellular locations and activities. , 2000, Journal of cell science.

[16]  G. Rustin,et al.  Role of tumour markers in monitoring epithelial ovarian cancer , 2000, British Journal of Cancer.

[17]  M. Mutoh,et al.  Copper-transporting P-type adenosine triphosphatase (ATP7B) is associated with cisplatin resistance. , 2000, Cancer research.

[18]  M. Morgan,et al.  Clinical characteristics of clear cell carcinoma of the ovary. , 1998, Gynecologic oncology.

[19]  Y. Minagawa,et al.  Glutathione concentration may be a useful predictor of response to second‐line chemotherapy in patients with ovarian cancer , 1998, Cancer.

[20]  H. Hidaka,et al.  A newly synthesized bifunctional inhibitor, CKA1083, enhances adriamycin activity against human ovarian carcinoma cells. , 1997, Anticancer Research.

[21]  Y. Minagawa,et al.  Glutathione S‐transferase‐π expression and glutathione concentration in ovarian carcinoma before and after chemotherapy , 1997 .

[22]  E. Partridge,et al.  Cyclophosphamide and cisplatin compared with paclitaxel and cisplatin in patients with stage III and stage IV ovarian cancer , 1996, The New England journal of medicine.

[23]  F. Kikkawa,et al.  Establishment and Characterization of Acquired Resistance to Platinum Anticancer Drugs in Human Ovarian Carcinoma Cells , 1995, Japanese journal of cancer research : Gann.

[24]  T. Ishikawa,et al.  Glutathione-associated cis-diamminedichloroplatinum(II) metabolism and ATP-dependent efflux from leukemia cells. Molecular characterization of glutathione-platinum complex and its biological significance. , 1993, The Journal of biological chemistry.

[25]  H. Hidaka,et al.  A newly synthesized bifunctional inhibitor, W-77, enhances adriamycin activity against human ovarian carcinoma cells. , 1993, Cancer research.

[26]  T. Mantle,et al.  Glutathione S-transferases. , 1990, Biochemical Society transactions.

[27]  D. Barton,et al.  Comparison of nonerythroid alpha-spectrin genes reveals strict homology among diverse species , 1988, Molecular and cellular biology.

[28]  H. Schägger,et al.  Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. , 1987, Analytical biochemistry.

[29]  R. Moon,et al.  cDNA cloning, sequencing and chromosome mapping of a non-erythroid spectrin, human α-fodrin , 1987 .

[30]  E. Yordan,et al.  A randomized trial of cyclophosphamide and doxorubicin with or without cisplatin in advanced ovarian carcinoma. A gynecologic oncology group study , 1986, Cancer.

[31]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[32]  W B Jakoby,et al.  Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. , 1974, The Journal of biological chemistry.

[33]  H. Kajiyama,et al.  Twist expression in patients with cervical cancer is associated with poor disease outcome. , 2008, Annals of oncology : official journal of the European Society for Medical Oncology.

[34]  J. Moscow,et al.  Glutathione-related enzymes, glutathione and multidrug resistance , 2004, Cytotechnology.

[35]  E. Heerkens,et al.  Identification of a novel C-terminal variant of β II spectrin : two isoforms of β II spectrin have distinct intracellular locations and activities , 2000 .

[36]  F. Kikkawa,et al.  Glutathione related enzymes in cis-diamminedichloroplatinum (II)-sensitive and-resistant human ovarian carcinoma cells. , 1994, Anticancer research.

[37]  R. Moon,et al.  cDNA cloning, sequencing and chromosome mapping of a non-erythroid spectrin, human alpha-fodrin. , 1987, Differentiation; research in biological diversity.