Silencing S100A4 gene promoted platinum sensitivity in ovarian cancer

Objective: To explore the relationship of S100A4 and platinum resistance in ovarian cancer. Materials and Methods: S100A4 expression level in tissue slices from 35 cisplatin-sensitive ovarian cancer patients and 28 cispatin-resistant patients were analyzed by immunohistochemistry. Platinum-resistant cells CP70 transfected with S100A4 siRNA were set as experimental group, negative control siRNA, and transfection reagent was also transfected cells and set as negative control group and blank group, respectively. After transfection, S100A4 transcription level of cells was determined by qRT-PCR, the expression levels of S100A4 and PKC/p53 pathway associated proteins were determined by Western-blot; cells ‘cisplatin sensitivity and cells’ invasion ability were analyzed by MTT method and Transwell assay and the cell cycle and apoptosis was determined by flow cytometry. Results: S100A4 expression level of cisplatinsensitive patients was lower than cisplatin-resistant patients; after transfection, IC₅₀ of blank group, negative control group and experimental group was 75.96, 66.73 and 40.31 μmol/L, respectively; the cells’ percentage in G1 phase of experimental group was higher than other groups while in S phase it was lower than other groups (p < 0.05); the expression level of PKC-δ/ε and p-p53 increased significantly (p < 0.05). Conclusions: Silencing S100A4 gene in CP70 cells could u-regulate the expression of PKC-δ/ε and p-p53, thus promoting apoptosis by PKC-δ/ε in coordination with p-p53 and increasing chemosensitivity of cells.

Ovarian cancer; S100A4 siRNA; Platinum resistance; Cisplatin sensitivity.

[1] Srisuttayasathien M., Khemapech N.: “Quality of life in ovarian cancer patients choosing to receive salvage chemotherapy or palliative treatment”. Asian Pac. J. Cancer Prev., 2013, 14, 7669.

[2] Li Y., Xiao S., Dan L., Xue M.: “P16INK4A is required for cisplatin resistance in vervical carcinoma SiHa cells”. Oncol. Lett., 2015, 9, 1104.

[3] Heizmann C.W., Cox J.A.: “New perspectives on S100 proteins: a multifunctional Ca(2+)-, Zn(2+)- and Cu(2+)-binding protein family”. Biometals, 1998, 11, 383.

[4] Schneider M., Hansen J.L., Sheikh S.P.: “S100A4: a common mediator of epithelial-mesenchymal transition, fibrosis and regeneration in diseases?” J. Mol. Med., 2008, 86, 507.

[5] Wang Y.Y., Ye Z.Y., Zhao Z.S., Tao H.Q., Chu Y.Q.: “High-level expression of S100A4 correlates with lymph node metastasis and poor prognosis in patients with gastric cancer”. Ann. Surg. Oncol., 2010, 17, 89.

[6] Wang H.Y., Zhang J.Y., Cui J.T., Tan X.H., Li W.M., Gu J., et al.: “Expression status of S100A14 and S100A4 correlates with metastatic potential and clinical outcome in colorectal cancer after surgery”. Oncol. Rep., 2010, 23, 45.

[7] Ismail N.I., Kaur G., Hashim H., Hassan M.S.: “S100A4 overexpression proves to be independent marker for breast cancer progression”. Cancer Cell. Int., 2008, 8, 12.

[8] Liang J., Piao Y., Holmes L., Fuller G.N., Henry V., Tiao N., de Groot J.F.: “Neutrophils promote the malignant glioma phenotype through S100A4”. Clin. Cancer Res., 2014, 20, 187.

[9] Mencia N., Selga E., Rico I., de Almagro M.C., Villalobos X., Ramirez S., et al.: “Overexpression of S100A4 in human cancer cell lines resistant to methothexate’. BMC Cancer, 2010, 10, 250.

[10] Kikuchi N., Horiuchi A., Osada R., Imai T., Wang C., Chen X., Konishi I.: “Nuclear expression of S100A4 is associated with aggressive behavior of epithelial ovarian carcinoma: an important autocrine/ paracrine factor in tumor progression”. J. Cancer Sci., 2006, 97, 1061.

[11] Kim Y.S., Hwan J.D., Bae S., Bae D.H., Shick W.A.: “Identification of differentially expressed genes using an annealing control primer system in stage III serous ovarian carcinoma”. BMC Cancer, 2010, 10, 576.

[12] Yu T., Yang Y., Zhang J., He H., Ren X.: “Circumvention of cisplatin resistance in ovarian cancer by combination o fcyclosporin A and low-intensity ultrasound”. Eur. J. Pharm. Biopharm., 2015, 91, 103.

[13] Stefanou D.T., Bamias A., Episkopou H., Kyrtopoulos S.A., Likka M., Kalampokas T., et al.: “Aberrant DNA damage response pathways may predict the outcome of platinum chemotherapy in ovarian cancer”. PLoS One, 2015, 10, e0117654.

[14] Zeisberg E.M., Potenta S., Xie L., Zeisberg M., Kalluri R.: “Discovery of endothelial to mesnchymal transition as a source for carcinoma- associated firbroblasts”. Cancer Res., 2007, 67, 10123.

[15] Zeisberg M., Hanai J., Sugimoto H., Mammoto T., Charytan D., Strutz F., et al.: “BMP-7 counteracts TGF-beta1-induced epithelialto- mesenchymal transition and reverses chronic renal injury”. Nat. Med., 2003, 9, 964.

[16] Liang D.P., Huang T.Q., Li S.J., Chen Z.J.: “Knockdown of S100A4 chemosensitizes human laryngeal carcinoma cells in vitro through inhibition of Slug”. Eur. Rev. Med. Pharmacol. Sci., 2014, 18, 3484.

[17] Basu A., Woolard M.D., Johnson C.L.: “Involvement of protein kinase C-delta in DNA damage-induced apoptosis”. Cell Death Differ., 2001, 8, 899.

[18] O’Brian C.A.: “Protein kinase C-alpha: a novel target for the therapy of androgen-independent prostate cancer? (Review-hypothesis)”. Oncol. Rep., 1998, 5, 305.

[19] Kenzelmann Broz D., Spano Mello S., Bieging K.T., Jiang D., Dusek R.L., Brady C.A., et al.: “Global genomic profiling reveals an extensive p53-regulated autophagy program contributing to key p53 responses”. Genes Dev., 2013, 27, 1016.

[20] Kim M.J., Kim R.K., Yoon C.H., An S., Hwang S.G., Suh Y., et al.: “Importance of PKCδ signaling in fractionated-radiation-induced expression of glioma-initiating cells and resistance to cancer treatment”. J. Cell. Sci., 2011, 124, 3084.

[21] Bredel M.: “Anticancer drug resistance in primary human brain tumors”. Brain Res. Brain Res. Rev., 2001, 35, 161.

[22] Wu H.M., Schally A.V., Cheng J.C., Zarandi M., Varga J., Leung P.C.: “Growth hormone-releasing hormone antagonist induces apoptosis of human endometrial cancer cells through PKCδ-mediated activation of p53/p21”. Cancer Lett., 2010, 298, 16.

[23] Albihn A., Mo H., Yang Y., Hennriksson M.: “Camptothecin-induced apoptosis is enhanced by Myc and involves PKCdelta signaling”. Int. J. Cancer, 2007, 121, 1821.

[24] Iioka Y., Mishima K., Azuma N., Tsuchida A., Takagi Y., Aoki T., Saito I.: “Overexpression of protein kinase C delta enhances displatin- induced cytotoxicity correlated with p53 in gastric cancer cell line”. Pathobiology, 2005, 72, 152.

[25] Meyer E., Vollmer J.Y., Bovey R., Stamenkovic I.: “Matrix metalloproteiases 9 and 10 inhibit protein kinase C-potentiated, p53-mediated apoptosis”. Cancer Res., 2005, 65, 4261.