Korean J Physiol Pharmacol.  2016 May;20(3):297-304. 10.4196/kjpp.2016.20.3.297.

Klotho plays a critical role in clear cell renal cell carcinoma progression and clinical outcome

• Kim JH
• Hwang KH
• Lkhagvadorj S
• Jung JH
• Chung HC
• Park KS
• Kong ID
• Eom M
• Cha SK

• KMID: 2285602
• DOI: http://doi.org/10.4196/kjpp.2016.20.3.297

Abstract

Klotho functions as a tumor suppressor predominantly expressed in renal tubular cells, the origin of clear cell renal cell carcinoma (ccRCC). Altered expression and/or activity of growth factor receptor have been implicated in ccRCC development. Although Klotho suppresses a tumor progression through growth factor receptor signaling including insulin-like growth factor-1 receptor (IGF-1R), the role of Klotho acting on IGF-1R in ccRCC and its clinical relevance remains obscure. Here, we show that Klotho is favorable prognostic factor for ccRCC and exerts tumor suppressive role for ccRCC through inhibiting IGF-1R signaling. Our data shows the following key findings. First, in tumor tissues, the level of Klotho and IGF-1R expression are low or high, respectively, compared to that of adjacent non-neoplastic parenchyma. Second, the Klotho expression is clearly low in higher grade of ccRCC and is closely associated with clinical outcomes in tumor progression. Third, Klotho suppresses IGF-1-stimulated cell proliferation and migration by inhibiting PI3K/Akt pathway. These results provide compelling evidence supporting that Klotho acting on IGF-1R signaling functions as tumor suppressor in ccRCC and suggest that Klotho is a potential carcinostatis substance for ccRCC.

Keyword

Clear cell renal cell carcinoma; IGF-1 receptor; Klotho; Migration; Prognosis

MeSH Terms

Carcinoma, Renal Cell*
Cell Proliferation
Prognosis
Receptor, IGF Type 1
Receptor, IGF Type 1

Figure

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  Fig. 1 Expression of Klotho and IGF-1R and clinical outcome.(A) Western blot analysis of expression of Klotho, p-IGF-1R and IGF-1R in tumor and adjacent non-tumor tissues of ccRCC. (B~E) Representative immunoblotting of Klotho (B), and IGF-1R and p-IGF-1R (D) in low and high nuclear grades of ccRCC. β-actin was used for loading control. (C and E) Densitometry from panel B and D, respectively (Rel.=relative). *p<0.05 versus low. N.S., not significant (F) IHC staining of Klotho and IGF-1R in low and high nuclear grade ccRCC. (G and H) Association between tumor size and expression levels of Klotho and IGF-1R, respectively. **p<0.01 versus high nuclear grade.

• 

  Fig. 2 Klotho suppresses serum growth factors and IGF-1-stimulated cell migration and proliferation of ccRCC via PI3K/Akt pathway.(A and B) Caki1 cell migration was examined using wound-healing assay with the presence of mitomycin C (0.1 µg/ml). Serum (10% FBS) and IGF-1 (100 nM)-induced cell migration was inhibited by pretreatment of Klotho (KL; 1 nM) in Caki1. (C) Number (No.) of migrated cells in the wound area in panel A and B. **p<0.01 versus vehicle (Veh). (D and E) Effect of Klotho on serum and IGF-1-induced colony formation in Caki1 cells. (F) Quantitative analysis of colony formation assay from three independent experiments in panel D and E. **p<0.01 versus Veh (G and I) Representative immunoblotting of p-Akt and p-Erk1/2 by IGF-1R stimulation and Klotho effect. GAPDH was used for loading control. (H and J) Quantitative analysis from panel G and I, respectively (Rel.= elative). **p<0.01 versus Veh. #p<0.01 versus no IGF-1. N.S., not significant. Note; in panel J, p-values for Veh vs KL are 0.13 and 0.08 in control (no IGF-1) and IGF-1-treated groups, respectively.

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