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Fibroblast growth factor 19 expression correlates with tumor progression and poorer prognosis of hepatocellular carcinoma.

Miura S, Mitsuhashi N, Shimizu H, Kimura F, Yoshidome H, Otsuka M, Kato A, Shida T, Okamura D, Miyazaki M - BMC Cancer (2012)

Bottom Line: We found that FGF19 was significantly overexpressed in HCCs as compared with corresponding noncancerous liver tissue (P < 0.05).Inversely, decreasing FGF19 and FGFR4 expression by siRNA significantly inhibited proliferation and increased apoptosis in JHH7 cells (P < 0.01, n = 12).The postoperative serum FGF19 levels in HCC patients was significantly lower than the preoperative levels (P < 0.01, n = 29).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1, Inohana, Chuo-ku, Chiba 260-0856, Japan.

ABSTRACT

Background: Although fibroblast growth factor 19 (FGF19) can promote liver carcinogenesis in mice, its involvement in human hepatocellular carcinoma (HCC) has not been well investigated. FGF19, a member of the FGF family, has unique specificity for its receptor FGFR4. This study aimed to clarify the involvement of FGF19 in the development of HCC.

Methods: We investigated human FGF19 and FGFR4 expression in 40 hepatocellular carcinoma specimens using quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) analysis and immunohistochemistry. Moreover, we examined the expression and the distribution of FGF19 and FGFR4 in 5 hepatocellular carcinoma cell lines (HepG2, HuH7, HLE, HLF, and JHH7) using RT-PCR and immunohistochemistry. To test the role of the FGF19/FGFR4 system in tumor progression, we used recombinant FGF19 protein and small interfering RNA (siRNA) of FGF19 and FGFR4 to regulate their concentrations.

Results: We found that FGF19 was significantly overexpressed in HCCs as compared with corresponding noncancerous liver tissue (P < 0.05). Univariate and multivariate analyses revealed that the tumor FGF19 mRNA expression was an independent prognostic factor for overall and disease-free survival. Moreover, we found that the FGF19 recombinant protein could increase the proliferation (P < 0.01, n = 12) and invasion (P < 0.01, n = 6) capabilities of human hepatocellular carcinoma cell lines and inhibited their apoptosis (P < 0.01, n = 12). Inversely, decreasing FGF19 and FGFR4 expression by siRNA significantly inhibited proliferation and increased apoptosis in JHH7 cells (P < 0.01, n = 12). The postoperative serum FGF19 levels in HCC patients was significantly lower than the preoperative levels (P < 0.01, n = 29).

Conclusions: FGF19 is critically involved in the development of HCCs. Targeting FGF19 inhibition is an attractive potential therapeutic strategy for HCC.

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(A) Proliferation changes after addition of various concentration of the FGF19 recombinant protein. (B) The proliferation index (PI) was defined as the OD values of cells treated with recombinant protein divided by those of untreated cells. The PI increased after addition of FGF19 recombinant protein (0.01-10 ng/mL final concentration) to culture media and culturing for 96 h. PI, proliferation index.
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Figure 4: (A) Proliferation changes after addition of various concentration of the FGF19 recombinant protein. (B) The proliferation index (PI) was defined as the OD values of cells treated with recombinant protein divided by those of untreated cells. The PI increased after addition of FGF19 recombinant protein (0.01-10 ng/mL final concentration) to culture media and culturing for 96 h. PI, proliferation index.

Mentions: We chose the HuH7, HepG2, HLE, HLF, and JHH7 cell lines upon consideration of the combination of FGF19 and FGFR4 levels. We cultured these HCC lines with FGF19 recombinant protein at concentrations of 0.01, 0.1, 0.5, 1, 5, 10, 50, or 100 ng/mL and performed a proliferation assay (Figure 4A; n = 12, P < 0.05). We found that the proliferation of all HCC cells examined increased significantly upon addition of FGF19 recombinant protein at concentrations of 0.01-10 ng/mL over 48-96 h (Figure 4B). The proliferation index was highest when the concentration of FGF19 was 1 ng/mL in the 5 HCC lines, whereas apoptosis of HCC cells was significantly suppressed by addition of 1 ng/mL FGF19 recombinant protein in culture media (Figure 5A; n = 12, P < 0.05). Tumor metastasis is still a major problem in management of cancer. In order to investigate further whether FGF19 plays an important role in tumor metastasis, the invasion assay and migration assay were performed. Invasion assays were used to investigate the alteration of cancer cell invasiveness in the presence of FGF19 recombinant protein for JHH7. The assays were conducted by staining and measuring the OD at 560 nm; they revealed that 1 ng/mL of the FGF19 recombinant protein significantly increased the mobility and invasiveness of the JHH7 cells (Figure 5B; n = 6, P < 0.05). As shown in Figure 5C, the number of migrated cells also increased dramatically after addition of the FGF19 recombinant protein (Figure 5C; n = 6, P < 0.05). Results of the invasion assay and migration assay indicate that FGF19 may be an activator of tumor metastasis.


Fibroblast growth factor 19 expression correlates with tumor progression and poorer prognosis of hepatocellular carcinoma.

Miura S, Mitsuhashi N, Shimizu H, Kimura F, Yoshidome H, Otsuka M, Kato A, Shida T, Okamura D, Miyazaki M - BMC Cancer (2012)

(A) Proliferation changes after addition of various concentration of the FGF19 recombinant protein. (B) The proliferation index (PI) was defined as the OD values of cells treated with recombinant protein divided by those of untreated cells. The PI increased after addition of FGF19 recombinant protein (0.01-10 ng/mL final concentration) to culture media and culturing for 96 h. PI, proliferation index.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3293719&req=5

Figure 4: (A) Proliferation changes after addition of various concentration of the FGF19 recombinant protein. (B) The proliferation index (PI) was defined as the OD values of cells treated with recombinant protein divided by those of untreated cells. The PI increased after addition of FGF19 recombinant protein (0.01-10 ng/mL final concentration) to culture media and culturing for 96 h. PI, proliferation index.
Mentions: We chose the HuH7, HepG2, HLE, HLF, and JHH7 cell lines upon consideration of the combination of FGF19 and FGFR4 levels. We cultured these HCC lines with FGF19 recombinant protein at concentrations of 0.01, 0.1, 0.5, 1, 5, 10, 50, or 100 ng/mL and performed a proliferation assay (Figure 4A; n = 12, P < 0.05). We found that the proliferation of all HCC cells examined increased significantly upon addition of FGF19 recombinant protein at concentrations of 0.01-10 ng/mL over 48-96 h (Figure 4B). The proliferation index was highest when the concentration of FGF19 was 1 ng/mL in the 5 HCC lines, whereas apoptosis of HCC cells was significantly suppressed by addition of 1 ng/mL FGF19 recombinant protein in culture media (Figure 5A; n = 12, P < 0.05). Tumor metastasis is still a major problem in management of cancer. In order to investigate further whether FGF19 plays an important role in tumor metastasis, the invasion assay and migration assay were performed. Invasion assays were used to investigate the alteration of cancer cell invasiveness in the presence of FGF19 recombinant protein for JHH7. The assays were conducted by staining and measuring the OD at 560 nm; they revealed that 1 ng/mL of the FGF19 recombinant protein significantly increased the mobility and invasiveness of the JHH7 cells (Figure 5B; n = 6, P < 0.05). As shown in Figure 5C, the number of migrated cells also increased dramatically after addition of the FGF19 recombinant protein (Figure 5C; n = 6, P < 0.05). Results of the invasion assay and migration assay indicate that FGF19 may be an activator of tumor metastasis.

Bottom Line: We found that FGF19 was significantly overexpressed in HCCs as compared with corresponding noncancerous liver tissue (P < 0.05).Inversely, decreasing FGF19 and FGFR4 expression by siRNA significantly inhibited proliferation and increased apoptosis in JHH7 cells (P < 0.01, n = 12).The postoperative serum FGF19 levels in HCC patients was significantly lower than the preoperative levels (P < 0.01, n = 29).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1, Inohana, Chuo-ku, Chiba 260-0856, Japan.

ABSTRACT

Background: Although fibroblast growth factor 19 (FGF19) can promote liver carcinogenesis in mice, its involvement in human hepatocellular carcinoma (HCC) has not been well investigated. FGF19, a member of the FGF family, has unique specificity for its receptor FGFR4. This study aimed to clarify the involvement of FGF19 in the development of HCC.

Methods: We investigated human FGF19 and FGFR4 expression in 40 hepatocellular carcinoma specimens using quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) analysis and immunohistochemistry. Moreover, we examined the expression and the distribution of FGF19 and FGFR4 in 5 hepatocellular carcinoma cell lines (HepG2, HuH7, HLE, HLF, and JHH7) using RT-PCR and immunohistochemistry. To test the role of the FGF19/FGFR4 system in tumor progression, we used recombinant FGF19 protein and small interfering RNA (siRNA) of FGF19 and FGFR4 to regulate their concentrations.

Results: We found that FGF19 was significantly overexpressed in HCCs as compared with corresponding noncancerous liver tissue (P < 0.05). Univariate and multivariate analyses revealed that the tumor FGF19 mRNA expression was an independent prognostic factor for overall and disease-free survival. Moreover, we found that the FGF19 recombinant protein could increase the proliferation (P < 0.01, n = 12) and invasion (P < 0.01, n = 6) capabilities of human hepatocellular carcinoma cell lines and inhibited their apoptosis (P < 0.01, n = 12). Inversely, decreasing FGF19 and FGFR4 expression by siRNA significantly inhibited proliferation and increased apoptosis in JHH7 cells (P < 0.01, n = 12). The postoperative serum FGF19 levels in HCC patients was significantly lower than the preoperative levels (P < 0.01, n = 29).

Conclusions: FGF19 is critically involved in the development of HCCs. Targeting FGF19 inhibition is an attractive potential therapeutic strategy for HCC.

Show MeSH
Related in: MedlinePlus