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Inactivation of bone morphogenetic protein 2 may predict clinical outcome and poor overall survival for renal cell carcinoma through epigenetic pathways.

Mitsui Y, Hirata H, Arichi N, Hiraki M, Yasumoto H, Chang I, Fukuhara S, Yamamura S, Shahryari V, Deng G, Saini S, Majid S, Dahiya R, Tanaka Y, Shiina H - Oncotarget (2015)

Bottom Line: The prevalence of BMP-2 promoter methylation was significantly greater and BMP-2 mRNA expression was significantly lower in RCC samples as compared to normal kidney samples.Furthermore, a significant correlation was found between BMP-2 promoter methylation and mRNA transcription in tumors.Aberrant BMP-2 methylation and the resultant loss of BMP-2 expression may be a useful molecular marker for designing improved diagnostic and therapeutic strategies for RCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Shimane University Faculty of Medicine, Enya-cho, Izumo, Japan.

ABSTRACT
We investigated whether impaired regulation of bone morphogenetic protein-2 (BMP-2) via epigenetic pathways is associated with renal cell carcinoma (RCC) pathogenesis. Expression and CpG methylation of the BMP-2 gene were analyzed using RCC cell lines, and 96 matched RCC and normal renal tissues. We also performed functional analysis using BMP-2 restored RCC cells. A significant association of BMP-2 mRNA expression was also found with advanced tumor stage and lymph node involvement, while lower BMP-2 mRNA expression was significantly associated with poor overall survival after radical nephrectomy. In RCC cells, BMP-2 restoration significantly inhibited cell proliferation, migration, invasion, and colony formation. In addition, BMP-2 overexpression induced p21(WAF1/CIP1) and p27(KIP1) expression, and cellular apoptosis in RCC cells. BMP-2 mRNA expression was significantly enhanced in RCC cells by 5-aza-2'-deoxycitidine treatment. The prevalence of BMP-2 promoter methylation was significantly greater and BMP-2 mRNA expression was significantly lower in RCC samples as compared to normal kidney samples. Furthermore, a significant correlation was found between BMP-2 promoter methylation and mRNA transcription in tumors. Aberrant BMP-2 methylation and the resultant loss of BMP-2 expression may be a useful molecular marker for designing improved diagnostic and therapeutic strategies for RCC.

No MeSH data available.


Related in: MedlinePlus

Schema of BMP-2/Smad signaling pathway in RCCBMP-2 expression is regulated by DNA promoter methylation. BMP-2 may cause the induction of GADD45a, p21 and p27 expression through the activation of Smad pathway. GADD45a will induce apoptosis via caspase cascade. In addition, p21 and p27 will induce cell cycle arrest by inhibiting CDK2. These cell cycle arrest and apoptotic effect may play an important role in the inhibition of tumor proliferation in RCC.
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Figure 7: Schema of BMP-2/Smad signaling pathway in RCCBMP-2 expression is regulated by DNA promoter methylation. BMP-2 may cause the induction of GADD45a, p21 and p27 expression through the activation of Smad pathway. GADD45a will induce apoptosis via caspase cascade. In addition, p21 and p27 will induce cell cycle arrest by inhibiting CDK2. These cell cycle arrest and apoptotic effect may play an important role in the inhibition of tumor proliferation in RCC.

Mentions: Since BMP-2 restoration significantly inhibited proliferation, migration, invasion, and colony formation in RCC cell lines, we hypothesized that its expression may induce apoptosis. The results of apoptosis assays of Caki-1 and Caki-2 cells performed 24 hours post-transfection are shown in Fig. 6A and B. Apoptotic and early apoptotic fractions (upper right and lower right, respectively, in quadrant images) were significantly greater in BMP-2 transfectants as compared to the vector control. These differences were also seen in both Caki-1 and Caki-2 cells at 48 hours after transfection (data not shown). These findings indicate a pro-apoptotic role for BMP-2, as well as its effects on the apoptotic pathway and regulation of tumorigenicity. In our recent study, we found that growth arrest and DNA damage inducible gene 45α (GADD45α) may play important roles in human RCC apoptosis [26]. Therefore, we examined the expression of several apoptotic proteins and GADD45α protein using Western blot analysis. As shown in Fig. 6C, BMP-2 over-expression caused an increase in cleaved caspase-3 in both Caki-1 and Caki-2 cells, further supporting the pro-apoptotic role of BMP-2. Furthermore, GADD45α was up-regulated, indicating that overexpression of BMP-2 induced GADD45α and apoptotic effects. Fig. 7 indicates the putative BMP-2 pathway in RCC on the basis of our results and previous studies [12, 13, 25, 27].


Inactivation of bone morphogenetic protein 2 may predict clinical outcome and poor overall survival for renal cell carcinoma through epigenetic pathways.

Mitsui Y, Hirata H, Arichi N, Hiraki M, Yasumoto H, Chang I, Fukuhara S, Yamamura S, Shahryari V, Deng G, Saini S, Majid S, Dahiya R, Tanaka Y, Shiina H - Oncotarget (2015)

Schema of BMP-2/Smad signaling pathway in RCCBMP-2 expression is regulated by DNA promoter methylation. BMP-2 may cause the induction of GADD45a, p21 and p27 expression through the activation of Smad pathway. GADD45a will induce apoptosis via caspase cascade. In addition, p21 and p27 will induce cell cycle arrest by inhibiting CDK2. These cell cycle arrest and apoptotic effect may play an important role in the inhibition of tumor proliferation in RCC.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496240&req=5

Figure 7: Schema of BMP-2/Smad signaling pathway in RCCBMP-2 expression is regulated by DNA promoter methylation. BMP-2 may cause the induction of GADD45a, p21 and p27 expression through the activation of Smad pathway. GADD45a will induce apoptosis via caspase cascade. In addition, p21 and p27 will induce cell cycle arrest by inhibiting CDK2. These cell cycle arrest and apoptotic effect may play an important role in the inhibition of tumor proliferation in RCC.
Mentions: Since BMP-2 restoration significantly inhibited proliferation, migration, invasion, and colony formation in RCC cell lines, we hypothesized that its expression may induce apoptosis. The results of apoptosis assays of Caki-1 and Caki-2 cells performed 24 hours post-transfection are shown in Fig. 6A and B. Apoptotic and early apoptotic fractions (upper right and lower right, respectively, in quadrant images) were significantly greater in BMP-2 transfectants as compared to the vector control. These differences were also seen in both Caki-1 and Caki-2 cells at 48 hours after transfection (data not shown). These findings indicate a pro-apoptotic role for BMP-2, as well as its effects on the apoptotic pathway and regulation of tumorigenicity. In our recent study, we found that growth arrest and DNA damage inducible gene 45α (GADD45α) may play important roles in human RCC apoptosis [26]. Therefore, we examined the expression of several apoptotic proteins and GADD45α protein using Western blot analysis. As shown in Fig. 6C, BMP-2 over-expression caused an increase in cleaved caspase-3 in both Caki-1 and Caki-2 cells, further supporting the pro-apoptotic role of BMP-2. Furthermore, GADD45α was up-regulated, indicating that overexpression of BMP-2 induced GADD45α and apoptotic effects. Fig. 7 indicates the putative BMP-2 pathway in RCC on the basis of our results and previous studies [12, 13, 25, 27].

Bottom Line: The prevalence of BMP-2 promoter methylation was significantly greater and BMP-2 mRNA expression was significantly lower in RCC samples as compared to normal kidney samples.Furthermore, a significant correlation was found between BMP-2 promoter methylation and mRNA transcription in tumors.Aberrant BMP-2 methylation and the resultant loss of BMP-2 expression may be a useful molecular marker for designing improved diagnostic and therapeutic strategies for RCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Shimane University Faculty of Medicine, Enya-cho, Izumo, Japan.

ABSTRACT
We investigated whether impaired regulation of bone morphogenetic protein-2 (BMP-2) via epigenetic pathways is associated with renal cell carcinoma (RCC) pathogenesis. Expression and CpG methylation of the BMP-2 gene were analyzed using RCC cell lines, and 96 matched RCC and normal renal tissues. We also performed functional analysis using BMP-2 restored RCC cells. A significant association of BMP-2 mRNA expression was also found with advanced tumor stage and lymph node involvement, while lower BMP-2 mRNA expression was significantly associated with poor overall survival after radical nephrectomy. In RCC cells, BMP-2 restoration significantly inhibited cell proliferation, migration, invasion, and colony formation. In addition, BMP-2 overexpression induced p21(WAF1/CIP1) and p27(KIP1) expression, and cellular apoptosis in RCC cells. BMP-2 mRNA expression was significantly enhanced in RCC cells by 5-aza-2'-deoxycitidine treatment. The prevalence of BMP-2 promoter methylation was significantly greater and BMP-2 mRNA expression was significantly lower in RCC samples as compared to normal kidney samples. Furthermore, a significant correlation was found between BMP-2 promoter methylation and mRNA transcription in tumors. Aberrant BMP-2 methylation and the resultant loss of BMP-2 expression may be a useful molecular marker for designing improved diagnostic and therapeutic strategies for RCC.

No MeSH data available.


Related in: MedlinePlus