J Korean Med Sci.  2008 Feb;23(1):94-97. 10.3346/jkms.2008.23.1.94.

Granulocyte Macrophage Colony-Stimulating Factor May Modulate the Post-transcription Pathway of Interleukin-6 Expression in Prostate Carcinoma Cells

• Kim TH
• Oh SY
• Kim YS
• Lee MK
• Kim HS
• Myung SC

• KMID: 1786852
• DOI: http://doi.org/10.3346/jkms.2008.23.1.94

Abstract

Interleukin-6 (IL-6) can stimulate a variety of tumors including prostatic carcinoma. Research has recently shown that IL-6 may act to stimulate the progression of prostatic cancer. To date, little research has been performed to better understand the nature of granulocyte macrophage colony-stimulating factor (GM-CSF) and the expression of IL-6. The aim of this study was to evaluate the effects of GM-CSF on the expression of IL-6 in prostate cancer-3 (PC-3) cells. The bone-derived PC-3 cell line was used in this study. Reverse transcription polymerase chain reaction (RT-PCR) and real- time PCR were performed to detect IL-6 mRNA expression. The IL-6 protein was measured by enzyme-linked immunosorbent assay (ELISA) after treatment with hGM-CSF. Our data indicated that IL-6 mRNA expression did not increase after treatment with hGM-CSF in comparison to the control group. However, the expression of IL-6 protein was increased compared to the control group. GM-CSF may modulate the post-transcription pathway of IL-6 expression in prostate carcinoma cells. Our data suggest that GM-CSF may have a role in IL-6-mediated development of prostate cancer.

Keyword

Prostatic Neoplasms; Interleukin-6; Granulocyte-Macrophage Colony-Stimulating Factor

MeSH Terms

Cell Line, Tumor
Enzyme-Linked Immunosorbent Assay
Granulocyte-Macrophage Colony-Stimulating Factor/*pharmacology
Humans
Interleukin-6/*genetics
Male
Prostatic Neoplasms/*genetics
*Protein Processing, Post-Translational
RNA, Messenger/analysis
Reverse Transcriptase Polymerase Chain Reaction/methods

Figure

• 

  Fig. 1 Time course of the IL-6 mRNA expression by conventional RT-PCR in PC-3 cells. IL-6 mRNA expression was gradually increased in control groups (lanes 1-5). IL-6 mRNA expression was increased at 4 hr, decreased until 12 hr, and then increased at 24 and 48 hr after hGM-CSF treatment (lanes 6-10). M, molecular size marker; 1-5, control groups; 6-10, hGM-CSF-treated groups.

• 

  Fig. 2 Time course of the IL-6 mRNA expression by real-time PCR. The PC-3 cells were treated with hGM-CSF for the indicated times. The control samples were incubated for 48 hr. The IL-6 mRNA expression was not increased at specific incubation time.

• 

  Fig. 3 The average IL-6 concentration (ng/mL) and their standard deviation by the incubations period as determined by ELISA. The control samples were incubated for 48 hr. The IL-6 protein expression was significantly increased at 4, 8, 12, 24, and 48 hr after hGM-CSF treatment compared with the control group. *p<0.01.

Reference

1. National Statistical office. Republic of Korea. Korea Statistical Information System. Dec., 31, 2002. Available from: URL:http://kosis.nso.go.kr.

2. Akira S, Taga T, Kishimoto T. Interleukin-6 in biology and medicine. Adv Immunol. 1993. 54:1–78.

3. Tan D, Wu X, Hou M, Lee SO, Lou W, Wang J, Janarthan B, Nallapareddy S, Trump DL, Gao AC. Interleukin-6 polymorphism is associated with more aggressive prostate cancer. J Urol. 2005. 174:753–756.

4. Chung TD, Yu JJ, Spiotto MT, Bartkowski M, Simons JW. Characterization of the role of IL-6 in the progression of prostate cancer. Prostate. 1999. 38:199–207.

5. Offner FA, Obrist P, Stadlmann S, Feichtinger H, Klingler P, Herold M, Zwierzina H, Hittmair A, Mikuz G, Abendstein B, Zeimet A, Marth C. IL-6 secretion by human peritoneal mesothelial and ovarian cancer cells. Cytokine. 1995. 7:542–547.

6. Lee SO, Lou W, Hou M, de Miguel F, Gerber L, Gao AC. Interleukin-6 promotes androgen-independent growth in LNCaP human prostate cancer cells. Clin Cancer Res. 2003. 9:370–376.

7. Horwood NJ, Udagawa N, Elliott J, Grail D, Okamura H, Kurimoto M, Dunn AR, Martin T, Gillespie MT. Interleukin 18 inhibits osteoclast formation via T cell production of granulocyte macrophage colony-stimulating factor. J Clin Invest. 1998. 101:595–603.

8. Savarese DM, Valinski H, Quesenberry P, Savarese T. Expression and function of colony-stimulating factors and their receptors in human prostate carcinoma cell lines. Prostate. 1998. 34:80–91.

9. Small EJ, Reese DM, Um B, Whisenant S, Dixon SC, Figg WD. Therapy of advanced prostate cancer with granulocyte macrophage colony-stimulating factor. Clin Cancer Res. 1999. 5:1738–1744.

10. Rini BI, Weinberg V, Bok R, Small EJ. Prostate-specific antigen kinetics as a measure of the biologic effect of granulocyte-macrophage colony-stimulating factor in patients with serologic progression of prostate cancer. J Clin Oncol. 2003. 21:99–105.

11. Kim TH, Kim YS, Myung SC, Lee DH, Won EH, Lee SY. The prostaglandin E receptor agonists increase granulocyte macrophage colony-stimulating factor in prostate cancer cells. Korean J Urol. 2004. 45:1272–1278.

12. Nakata H, Uemura Y, Kobayashi M, Harada R, Taguchi H. Cyclooxygenase-2 inhibitor NS-398 suppresses cell growth and constitutive production of granulocyte-colony stimulating factor and granulocyte macrophage-colony stimulating factor in lung cancer cells. Cancer Sci. 2003. 94:173–180.

13. Guillaume T, Sekhavat M, Rubinstein DB, Hamdan O, Symann ML. Transcription of genes encoding granulocyte-macrophage colony-stimulating factor, interleukin 3, and interleukin 6 receptors and lack of proliferative response to exogenous cytokines in nonhematopoietic human malignant cell lines. Cancer Res. 1993. 53:3139–3144.

14. Rokhlin OW, Griebling TL, Karassina NV, Raines MA, Cohen MB. Human prostate carcinoma cell lines secrete GM-CSF and express GM-CSF-receptor on their cell surface. Anticancer Res. 1996. 16:557–563.

15. Markowicz S, Engleman EG. Granulocyte-macrophage-colony stimulating factor promotes differentiation and survival of human peripheral blood dendritic cells in vitro. J Clin Invest. 1990. 85:955–961.

16. Culig Z. Interleukin-6 polymorphism: expression and pleiotropic regulation in human prostate cancer. J Urol. 2005. 174:417.

17. Nakashima J, Tachibana M, Horiguchi Y, Oya M, Ohigashi T, Asakura H, Murai M. Serum interleukin 6 as a prognostic factor in patients with prostate cancer. Clin Cancer Res. 2000. 6:2702–2706.

18. Drachenberg DE, Elgamal AA, Rowbotham R, Peterson M, Murphy GP. Circulating levels of interleukin-6 in patients with hormone refractory prostate cancer. Prostate. 1999. 41:127–133.

19. Adler HL, McCurdy MA, Kattan MW, Timme TL, Scardino PT, Thompson TC. Elevated levels of circulating interleukin-6 and transforming growth factor-beta1 in patients with metastatic prostate carcinoma. J Urol. 1999. 161:182–187.

20. Okamoto M, Lee C, Oyasu R. Interleukin-6 as a paracrine and autocrine growth factor in human prostatic carcinoma cells in vitro. Cancer Res. 1997. 57:141–146.

21. Siegall CB, Schwab G, Nordan RP, FitzGerald DJ, Pastan I. Expression of the interleukin-6 receptor and interleukin-6 in prostate carcinoma cells. Cancer Res. 1990. 50:7786–7788.

22. Ivanovic V, Melman A, Davis-Joseph B, Valcic M, Geliebter J. Elevated plasma levels of TGF-beta 1 in patients with invasive prostate cancer. Nat Med. 1995. 1:282–284.

23. Charyulu VI, Lopez DM. Elevated GM-CSF levels in tumor bearing mice upregulate IL-6 production by B cells via a mechanism independent of TNF-alpha. Int J Oncol. 2000. 16:161–167.