Open Access

FOXO1 regulates oncogenic PKC-ι expression in melanoma inversely to c-Jun in an autocrine manner via IL-17E and ICAM-1 activation

  • Authors:
    • Wishrawana S. Ratnayke
    • Christopher A. Apostolatos
    • Sloan Breedy
    • Andre H. Apostolatos
    • Mildred Acevedo-Duncan
  • View Affiliations

  • Published online on: October 25, 2018     https://doi.org/10.3892/wasj.2018.2
  • Pages: 25-38
  • Copyright: © Ratnayke et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Regardless of abundant efforts to enhance primary prevention and early detection, the number of melanoma cases in the United States has increased steadily over the past 15 years, thus greatly affecting public health and the economy. In previous studies, we demonstrated protein kinase C‑ι (PKC‑ι) to be an oncogene in melanoma, which promotes the activation of nuclear factor (NF)‑κB, thereby supporting survival and progression. In addition, we demonstrated that PKC‑ι induced the metastasis of melanoma cells by activating Vimentin, and PKC‑ι inhibition downregulated epithilial‑mesencymal transi‑ tion (EMT), while inducing apoptosis. Of note, PKC‑ι specific inhibitors downregulated the expression of both PKC‑ι and phosphorylated PKC‑ι, suggesting that PKC‑ι plays a role in regulating its own expression in melanoma. In this study, we report the underlaying mechanisms of the transcriptional regulation of PKC‑ι (PRKCI gene) expression in melanoma. c‑Jun, interferon‑stimulated gene factor 3 (ISGF3), paired box gene 3 (PAX3), early growth response protein 1 (EGR1) and Forkhead box protein O1 (FOXO1), which bind on or near the promoter sequence of the PRKCI gene, were analyzed for their role in PKC‑ι regulation in SK‑MEL‑2 and MeWo cell lines. We silenced selected transcription factors using siRNA, and the results revealed that the silencing of c‑Jun and FOXO1 significantly altered the expression of PRKCI. The levels of both phosphorylated and total PKC‑ι increased upon FOXO1 silencing and decreased upon c‑Jun silencing, suggesting that c‑Jun acts as an upregulator, while FOXO1 acts as a down‑ regulator of PRKCI expression. We also used a multiplex ELISA to analyze multiple pathways other than NF‑κB that were affected by treatment with PKC‑ι inhibitor. The silencing of NF‑κB p65 and PKC‑ι by siRNA suggested that the regula‑ tion of PKC‑ι expression was strongly associated with FOXO1. In addition, we observed a significant decrease in the mRNA levels of both interleukin (IL)‑6 and IL‑8, with a significant increase in the levels of IL‑17E and intercellular adhesion molecule 1 (ICAM‑1) upon the knockdown of expression of PKC‑ι in both cell lines. This suggested that PKC‑ι expres‑ sion was affected by these cytokines in an autocrine manner. Overall, the findings of this study suggest that PKC‑ι inhibi‑ tion suppresses its own expression, diminishing oncogenic signaling, while upregulating anti‑tumor signaling, thus rendering it an effective novel biomarker for use in the design of novel targeted therapeutics for melanoma.

References

1 

Cronin KA, Lake AJ, Scott S, Sherman RL, Noone AM, Howlader N, Henley SJ, Anderson RN, Firth AU, Ma J, et al: Annual Report to the Nation on the Status of Cancer, part I: National cancer statistics. Cancer. 124:2785–2800. 2018.PubMed/NCBI View Article : Google Scholar

2 

Nazarian R, Shi H, Wang Q, Kong X, Koya RC, Lee H, Chen Z, Lee MK, Attar N, Sazegar H, et al: Melanomas acquire resistance to B-RAF(V600E) inhibition by RTK or N-RAS upregulation. Nature. 468:973–977. 2010.PubMed/NCBI View Article : Google Scholar

3 

Ratnayake WS, Apostolatos CA, Apostolatos AH, Schutte RJ, Huynh MA, Ostrov DA and Acevedo-Duncan M: Oncogenic PKC-ι activates Vimentin during epithelial-mesenchymal transition in melanoma; a study based on PKC-ι and PKC-ζ specific inhibitors. Cell Adhes Migr. 0:1–17. 2018.PubMed/NCBI View Article : Google Scholar

4 

Ratnayake WS, Apostolatos AH, Ostrov DA and Acevedo-Duncan M: Two novel atypical PKC inhibitors; ACPD and DNDA effectively mitigate cell proliferation and epithelial to mesenchymal transition of metastatic melanoma while inducing apoptosis. Int J Oncol. 51:1370–1382. 2017.PubMed/NCBI View Article : Google Scholar

5 

Ratnayake WS and Acevedo-Duncan M: Abstract 862: Atypical protein kinase c inhibitors can repress epithelial to mesenchymal transition (type III) in malignant melanoma. Cancer Res. 77((Suppl 13)): 862. 2017. View Article : Google Scholar

6 

Manning G, Whyte DB, Martinez R, Hunter T and Sudarsanam S: The protein kinase complement of the human genome. Science. 298:1912–1934. 2002.PubMed/NCBI View Article : Google Scholar

7 

Regala RP, Weems C, Jamieson L, Khoor A, Edell ES, Lohse CM and Fields AP: Atypical protein kinase C iota is an oncogene in human non-small cell lung cancer. Cancer Res. 65:8905–8911. 2005.PubMed/NCBI View Article : Google Scholar

8 

Murray NR and Fields AP: Atypical protein kinase C iota protects human leukemia cells against drug-induced apoptosis. J Biol Chem. 272:27521–27524. 1997.PubMed/NCBI View Article : Google Scholar

9 

Desai S, Pillai P, Win-Piazza H and Acevedo-Duncan M: PKC-ι promotes glioblastoma cell survival by phosphorylating and inhibiting BAD through a phosphatidylinositol 3-kinase pathway. Biochim Biophys Acta. 1813:1190–1197. 2011.PubMed/NCBI View Article : Google Scholar

10 

Win HY and Acevedo-Duncan M: Role of protein kinase C-iota in transformed non-malignant RWPE-1 cells and androgen-independent prostate carcinoma DU-145 cells. Cell Prolif. 42:182–194. 2009.PubMed/NCBI View Article : Google Scholar

11 

Eder AM, Sui X, Rosen DG, Nolden LK, Cheng KW, Lahad JP, Kango-Singh M, Lu KH, Warneke CL, Atkinson EN, et al: Atypical PKCiota contributes to poor prognosis through loss of apical-basal polarity and cyclin E overexpression in ovarian cancer. Proc Natl Acad Sci USA. 102:12519–12524. 2005.PubMed/NCBI View Article : Google Scholar

12 

Apostolatos AH, Ratnayake WS, Win-Piazza H, Apostolatos CA, Smalley T, Kang L, Salup R, Hill R and Acevedo-Duncan M: Inhibition of atypical protein kinase C-ι effectively reduces the malignancy of prostate cancer cells by downregulating the NF-κB signaling cascade. Int J Oncol. 53:1836–1846. 2018.PubMed/NCBI View Article : Google Scholar

13 

Wu J, Lu M, Li Y, Shang YK, Wang SJ, Meng Y, Wang Z, Li ZS, Chen H, Chen ZN, et al: Regulation of a TGF-β1-CD147 self-sustaining network in the differentiation plasticity of hepatocellular carcinoma cells. Oncogene. 35:5468–5479. 2016.PubMed/NCBI View Article : Google Scholar

14 

Venter JC, Adams MD, Myers EW, Li PW, Mural RJ, Sutton GG, Smith HO, Yandell M, Evans CA, Holt RA, et al: The sequence of the human genome. Science. 291:1304–1351. 2001.PubMed/NCBI View Article : Google Scholar

15 

Fagerberg L, Hallström BM, Oksvold P, Kampf C, Djureinovic D, Odeberg J, Habuka M, Tahmasebpoor S, Danielsson A, Edlund K, et al: Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics. Mol Cell Proteomics. 13:397–406. 2014.PubMed/NCBI View Article : Google Scholar

16 

Win HY and Acevedo-Duncan M: Atypical protein kinase C phosphorylates IKKalphabeta in transformed non-malignant and malignant prostate cell survival. Cancer Lett. 270:302–311. 2008.PubMed/NCBI View Article : Google Scholar

17 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar

18 

Butler AM, Scotti Buzhardt ML, Erdogan E, Li S, Inman KS, Fields AP and Murray NR: A small molecule inhibitor of atypical protein kinase C signaling inhibits pancreatic cancer cell transformed growth and invasion. Oncotarget. 6:15297–15310. 2015.PubMed/NCBI View Article : Google Scholar

19 

Wisdom R, Johnson RS and Moore C: c-Jun regulates cell cycle progression and apoptosis by distinct mechanisms. EMBO J. 18:188–197. 1999.PubMed/NCBI View Article : Google Scholar

20 

Angel P, Hattori K, Smeal T and Karin M: The jun proto-oncogene is positively autoregulated by its product, Jun/AP-1. Cell. 55:875–885. 1988.PubMed/NCBI View Article : Google Scholar

21 

Lopez-Bergami P, Huang C, Goydos JS, Yip D, Bar-Eli M, Herlyn M, Smalley KS, Mahale A, Eroshkin A, Aaronson S, et al: Rewired ERK-JNK signaling pathways in melanoma. Cancer Cell. 11:447–460. 2007.PubMed/NCBI View Article : Google Scholar

22 

Vogt PK: Fortuitous convergences: The beginnings of JUN. Nat Rev Cancer. 2:465–469. 2002.PubMed/NCBI View Article : Google Scholar

23 

Szabo E, Riffe ME, Steinberg SM, Birrer MJ and Linnoila RI: Altered cJUN expression: An early event in human lung carcinogenesis. Cancer Res. 56:305–315. 1996.PubMed/NCBI

24 

Vleugel MM, Greijer AE, Bos R, van der Wall E and van Diest PJ: c-Jun activation is associated with proliferation and angiogenesis in invasive breast cancer. Hum Pathol. 37:668–674. 2006.PubMed/NCBI View Article : Google Scholar

25 

Behrens A, Sibilia M and Wagner EF: Amino-terminal phosphorylation of c-Jun regulates stress-induced apoptosis and cellular proliferation. Nat Genet. 21:326–329. 1999.PubMed/NCBI View Article : Google Scholar

26 

Nateri AS, Spencer-Dene B and Behrens A: Interaction of phosphorylated c-Jun with TCF4 regulates intestinal cancer development. Nature. 437:281–285. 2005.PubMed/NCBI View Article : Google Scholar

27 

Rena G, Guo S, Cichy SC, Unterman TG and Cohen P: Phosphorylation of the transcription factor forkhead family member FKHR by protein kinase B. J Biol Chem. 274:17179–17183. 1999.PubMed/NCBI View Article : Google Scholar

28 

Nakae J, Kitamura T, Kitamura Y, Biggs WH III, Arden KC and Accili D: The forkhead transcription factor Foxo1 regulates adipocyte differentiation. Dev Cell. 4:119–129. 2003.PubMed/NCBI View Article : Google Scholar

29 

Matsuzaki H, Daitoku H, Hatta M, Tanaka K and Fukamizu A: Insulin-induced phosphorylation of FKHR (Foxo1) targets to proteasomal degradation. Proc Natl Acad Sci USA. 100:11285–11290. 2003.PubMed/NCBI View Article : Google Scholar

30 

Lu H and Huang H: FOXO1: A potential target for human diseases. Curr Drug Targets. 12:1235–1244. 2011.PubMed/NCBI View Article : Google Scholar

31 

Borkhardt A, Repp R, Haas OA, Leis T, Harbott J, Kreuder J, Hammermann J, Henn T and Lampert F: Cloning and characterization of AFX, the gene that fuses to MLL in acute leukemias with a t(X;11)(q13;q23). Oncogene. 14:195–202. 1997.PubMed/NCBI View Article : Google Scholar

32 

Anderson MJ, Viars CS, Czekay S, Cavenee WK and Arden KC: Cloning and characterization of three human forkhead genes that comprise an FKHR-like gene subfamily. Genomics. 47:187–199. 1998.PubMed/NCBI View Article : Google Scholar

33 

Zhang X, Tang N, Hadden TJ and Rishi AK: Akt, FoxO and regulation of apoptosis. Biochim Biophys Acta. 1813:1978–1986. 2011.PubMed/NCBI View Article : Google Scholar

34 

Farhan M, Wang H, Gaur U, Little PJ, Xu J and Zheng W: FOXO signaling pathways as therapeutic targets in cancer. Int J Biol Sci. 13:815–827. 2017.PubMed/NCBI View Article : Google Scholar

35 

Fu Z and Tindall DJ: FOXOs, cancer and regulation of apoptosis. Oncogene. 27:2312–2319. 2008.PubMed/NCBI View Article : Google Scholar

36 

Zhang Y, Zhang L, Sun H, Lv Q, Qiu C, Che X, Liu Z and Jiang J: Forkhead transcription factor 1 inhibits endometrial cancer cell proliferation via sterol regulatory element-binding protein 1. Oncol Lett. 13:731–737. 2017.PubMed/NCBI View Article : Google Scholar

37 

Hodge DR, Hurt EM and Farrar WL: The role of IL-6 and STAT3 in inflammation and cancer. Eur J Cancer. 41:2502–2512. 2005.PubMed/NCBI View Article : Google Scholar

38 

Yue P and Turkson J: Targeting STAT3 in cancer: How successful are we? Expert Opin Investig Drugs. 18:45–56. 2009.PubMed/NCBI View Article : Google Scholar

39 

Jing N and Tweardy DJ: Targeting Stat3 in cancer therapy. Anticancer Drugs. 16:601–607. 2005.PubMed/NCBI

40 

Page BDG, Khoury H, Laister RC, Fletcher S, Vellozo M, Manzoli A, Yue P, Turkson J, Minden MD and Gunning PT: Small molecule STAT5-SH2 domain inhibitors exhibit potent antileukemia activity. J Med Chem. 55:1047–1055. 2012.PubMed/NCBI View Article : Google Scholar

41 

Pardanani A, Lasho T, Smith G, Burns CJ, Fantino E and Tefferi A: CYT387, a selective JAK1/JAK2 inhibitor: In vitro assessment of kinase selectivity and preclinical studies using cell lines and primary cells from polycythemia vera patients. Leukemia. 23:1441–1445. 2009.PubMed/NCBI View Article : Google Scholar

42 

Rani A and Murphy JJ: STAT5 in cancer and immunity. J Interferon Cytokine Res. 36:226–237. 2016.PubMed/NCBI View Article : Google Scholar

43 

Korneev KV, Atretkhany KN, Drutskaya MS, Grivennikov SI, Kuprash DV and Nedospasov SA: TLR-signaling and proinflammatory cytokines as drivers of tumorigenesis. Cytokine. 89:127–135. 2017.PubMed/NCBI View Article : Google Scholar

44 

Zhang X, Wrzeszczynska MH, Horvath CM and Darnell JE Jr: Interacting regions in Stat3 and c-Jun that participate in cooperative transcriptional activation. Mol Cell Biol. 19:7138–7146. 1999.PubMed/NCBI View Article : Google Scholar

45 

Hornsveld M, Dansen TB, Derksen PW and Burgering BM: Re-evaluating the role of FOXOs in cancer. Semin Cancer Biol. 50:90–100. 2018.PubMed/NCBI View Article : Google Scholar

46 

Sunters A, Madureira PA, Pomeranz KM, Aubert M, Brosens JJ, Cook SJ, Burgering BM, Coombes RC and Lam EW: Paclitaxel-induced nuclear translocation of FOXO3a in breast cancer cells is mediated by c-Jun NH2-terminal kinase and Akt. Cancer Res. 66:212–220. 2006.PubMed/NCBI View Article : Google Scholar

47 

Yuan ZL, Guan YJ, Wang L, Wei W, Kane AB and Chin YE: Central role of the threonine residue within the p+1 loop of receptor tyrosine kinase in STAT3 constitutive phosphorylation in metastatic cancer cells. Mol Cell Biol. 24:9390–9400. 2004.PubMed/NCBI View Article : Google Scholar

48 

Antonicelli F, Lorin J, Kurdykowski S, Gangloff SC, Le Naour R, Sallenave JM, Hornebeck W, Grange F and Bernard P: CXCL10 reduces melanoma proliferation and invasiveness in vitro and in vivo. Br J Dermatol. 164:720–728. 2011.PubMed/NCBI View Article : Google Scholar

49 

Zaynagetdinov R, Sherrill TP, Gleaves LA, McLoed AG, Saxon JA, Habermann AC, Connelly L, Dulek D, Peebles RS Jr, Fingleton B, et al: Interleukin-5 facilitates lung metastasis by modulating the immune microenvironment. Cancer Res. 75:1624–1634. 2015.PubMed/NCBI View Article : Google Scholar

50 

Sun X, Cheng G, Hao M, Zheng J, Zhou X, Zhang J, Taichman RS, Pienta KJ and Wang J: CXCL12/CXCR4/CXCR7 chemokine axis and cancer progression. Cancer Metastasis Rev. 29:709–722. 2010.PubMed/NCBI View Article : Google Scholar

51 

Wightman SC, Uppal A, Pitroda SP, Ganai S, Burnette B, Stack M, Oshima G, Khan S, Huang X, Posner MC, et al: Oncogenic CXCL10 signalling drives metastasis development and poor clinical outcome. Br J Cancer. 113:327–335. 2015.PubMed/NCBI View Article : Google Scholar

52 

Ishiguro H, Akimoto K, Nagashima Y, Kojima Y, Sasaki T, Ishiguro-Imagawa Y, Nakaigawa N, Ohno S, Kubota Y and Uemura H: aPKClambda/ι promotes growth of prostate cancer cells in an autocrine manner through transcriptional activation of interleukin-6. Proc Natl Acad Sci USA. 106:16369–16374. 2009.PubMed/NCBI View Article : Google Scholar

53 

Peng H, Chen P, Cai Y, Chen Y, Wu QH, Li Y, Zhou R and Fang X: Endothelin-1 increases expression of cyclooxygenase-2 and production of interlukin-8 in hunan pulmonary epithelial cells. Peptides. 29:419–424. 2008.PubMed/NCBI View Article : Google Scholar

54 

Timani KA, Győrffy B, Liu Y, Mohammad KS and He JJ: Tip110/SART3 regulates IL-8 expression and predicts the clinical outcomes in melanoma. Mol Cancer. 17(124)2018.PubMed/NCBI View Article : Google Scholar

55 

Yang M, Liu J, Piao C, Shao J and Du J: ICAM-1 suppresses tumor metastasis by inhibiting macrophage M2 polarization through blockade of efferocytosis. Cell Death Dis. 6(e1780)2015.PubMed/NCBI View Article : Google Scholar

56 

de Groote ML, Kazemier HG, Huisman C, van der Gun BT, Faas MM and Rots MG: Upregulation of endogenous ICAM-1 reduces ovarian cancer cell growth in the absence of immune cells. Int J Cancer. 134:280–290. 2014.PubMed/NCBI View Article : Google Scholar

57 

Benatar T, Cao MY, Lee Y, Lightfoot J, Feng N, Gu X, Lee V, Jin H, Wang M, Wright JA, et al: IL-17E, a proinflammatory cytokine, has antitumor efficacy against several tumor types in vivo. Cancer Immunol Immunother. 59:805–817. 2010.PubMed/NCBI View Article : Google Scholar

58 

Benatar T, Cao MY, Lee Y, Li H, Feng N, Gu X, Lee V, Jin H, Wang M, Der S, et al: Virulizin induces production of IL-17E to enhance antitumor activity by recruitment of eosinophils into tumors. Cancer Immunol Immunother. 57:1757–1769. 2008.PubMed/NCBI View Article : Google Scholar

59 

Wei C, Sirikanjanapong S, Lieberman S, Delacure M, Martiniuk F, Levis W and Wang BY: Primary mucosal melanoma arising from the eustachian tube with CTLA-4, IL-17A, IL-17C, and IL-17E upregulation. Ear Nose Throat J. 92:36–40. 2013.PubMed/NCBI

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Copy and paste a formatted citation
APA
Ratnayke, W., Apostolatos, C., Breedy, S., Apostolatos, A., & Acevedo-Duncan, M. (2019). FOXO1 regulates oncogenic PKC-ι expression in melanoma inversely to c-Jun in an autocrine manner via IL-17E and ICAM-1 activation. World Academy of Sciences Journal, 1, 25-38. https://doi.org/10.3892/wasj.2018.2
MLA
Ratnayke, W., Apostolatos, C., Breedy, S., Apostolatos, A., Acevedo-Duncan, M."FOXO1 regulates oncogenic PKC-ι expression in melanoma inversely to c-Jun in an autocrine manner via IL-17E and ICAM-1 activation". World Academy of Sciences Journal 1.1 (2019): 25-38.
Chicago
Ratnayke, W., Apostolatos, C., Breedy, S., Apostolatos, A., Acevedo-Duncan, M."FOXO1 regulates oncogenic PKC-ι expression in melanoma inversely to c-Jun in an autocrine manner via IL-17E and ICAM-1 activation". World Academy of Sciences Journal 1, no. 1 (2019): 25-38. https://doi.org/10.3892/wasj.2018.2