Cancer Res Treat.  2021 Oct;53(4):1174-1183. 10.4143/crt.2021.031.

Ferroportin and FBXL5 as Prognostic Markers in Advanced Stage Clear Cell Renal Cell Carcinoma

• Park CK
• Heo J
• Ham WS
• Choi YD
• Shin SJ
• Cho NH

• KMID: 2521592
• DOI: http://doi.org/10.4143/crt.2021.031

Abstract

Purpose
Advanced stage clear cell renal cell carcinoma (ccRCC) involves a poor prognosis. Several studies have reported that dysfunctions in iron metabolism‒related proteins may cause tumor progression and metastasis of this carcinoma. In this study, we investigated the impact of the expression of iron metabolism‒related proteins on patient prognoses in advanced stage ccRCCs.
Materials and Methods
All of 143 advanced stage ccRCC specimens were selected following validation with double blind reviews. Several clinicopathological parameters including nuclear grade, perirenal fat invasion, renal sinus fat invasion, vascular invasion, necrosis, and sarcomatoid/rhabdoid differentiation were compared with the expression of ferroportin (FPN), and F-Box and leucine rich repeat protein 5 (FBXL5), by immunohistochemistry. FPN and FBXL5 mRNA level of ccRCC from The Cancer Genome Atlas database were also analyzed for validation.
Results
FPN and FBXL5 immunohistochemistry showed membrane and cytoplasmic expression, respectively. Based on the H-score, cases were classified as low or high expression with a cutoff value of 20 for FPN and 15 for FBXL5, respectively. Low expression of FPN and FBXL5 were significantly associated with patient death (p=0.022 and p=0.005, respectively). In survival analyses, low expression of FPN and FBXL5 were significantly associated with shorter overall survival (p=0.003 and p=0.004, respectively). On multivariate analysis, low expression of FBXL5 (hazard ratio, 2.001; p=0.034) was significantly associated with shorter overall survival.
Conclusion
FPN and FBXL5 can be used as potential prognostic markers and therapeutic targets for advanced stage ccRCC.

Keyword

Clear cell renal cell carcinoma; Ferroportin; F-Box and leucine rich repeat protein 5; Immunohistochemistry; Prognosis

Figure

• 

  Fig. 1 Representative cases of ferroportin (FPN), F-Box, and leucine rich repeat protein 5 (FBXL5) and signal transducer and activator of transcription 3 (STAT3) expression in clear cell renal cell carcinomas (×200). FPN showed membrane expression with nonspecific nuclear staining: (A) negative, (B) intensity 1+, (C) intensity 2+, and (D) intensity 3+. FBXL5 cytoplasmic expression: (E) negative, (F) intensity 1+, (G) intensity 2+, and (H) intensity 3+. STAT3 showed nuclear expression: (I) negative, (J) intensity 1+, (K) intensity 2+, and (L) intensity 3+.

• 

  Fig. 2 Overall survival (OS) of clear cell renal cell carcinoma patients according to ferroportin (FPN), F-Box, and leucine rich repeat protein 5 (FBXL5) and signal transducer and activator of transcription 3 (STAT3) expression. (A) In 143 clear cell renal cell carcinoma, low expression of FPN was significantly associated with shorter OS (p=0.003). (B) Similar to FPN, low FBXL5 expression was significantly associated with shorter OS (p=0.004). (C) Cases with low expression of both FPN and FBXL5 showed significantly shorter OS than other cases (p < 0.001). (D) High STAT3 expression showed significantly shorter OS (p=0.007). In patients with recurrence or metastasis, those with low FPN (E) and FBXL5 (F) expression showed significantly shorter OS (p=0.016 and p=0.015, respectively). (G) High STAT3 expression did not show significant differences in patients with recurrence or metastasis (p=0.525). (H) When analyzing 191 pT3 and pT4 patients of The Cancer Genome Atlas cohort, low mRNA expression level of FPN was significantly associated with shorter OS (p < 0.001). (I) Similar to FPN, the low mRNA expression level of FBXL5 was significantly associated with shorter OS (p=0.014).

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  Fig. 3 Schematic diagram of cross-talk between iron-related metabolism and hypoxia-inducible factor 1α (HIF-1α) in clear cell renal cell carcinoma. The cytosolic excess iron and hydrogen peroxide trigger the Fenton reaction with generation of reactive oxygen species, hydroxyl radicals, and hydroxide anions. To maintain the cytosolic and mitochondrial pH, glycolysis, lactate formation, and subsequent nucleotide synthesis is promoted. Inactivation of VHL induces the activation of HIF-1α, which promotes the expression of glycolysis-related genes and LDHA. FBXL5, F-Box and leucine rich repeat protein 5.

Reference

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