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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

Effects of BMP-2 overexpression on cell proliferation, migration, invasion, and colony-formation(A) BMP-2 expression level in RCC cell lines (Caki-1, Caki-2) were determined by immunoblotting analysis at 48 hours after transfection of a plasmid containing human BMP-2. (B) Cell viability was analyzed using an MTS cell proliferation assay at 24, 48, 72, and 96 hours after transient transfection. Overexpression of BMP-2 significantly inhibited cell viability. *P<0.05, **P<0.01. (C) Representative images from wound healing assay. After transfection (48 hours), a wound was formed by scraping, then measured 24 hours later. Over-expression of BMP-2 significantly inhibited cell migration. *P<0.0001. (D) Representative images from invasion assay. Overexpression of BMP-2 resulted in significantly decreased cell invasion. *P<0.0001. (E) Overexpression of BMP-2 significantly inhibited colony formation ability. *P<0.01.
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Figure 4: Effects of BMP-2 overexpression on cell proliferation, migration, invasion, and colony-formation(A) BMP-2 expression level in RCC cell lines (Caki-1, Caki-2) were determined by immunoblotting analysis at 48 hours after transfection of a plasmid containing human BMP-2. (B) Cell viability was analyzed using an MTS cell proliferation assay at 24, 48, 72, and 96 hours after transient transfection. Overexpression of BMP-2 significantly inhibited cell viability. *P<0.05, **P<0.01. (C) Representative images from wound healing assay. After transfection (48 hours), a wound was formed by scraping, then measured 24 hours later. Over-expression of BMP-2 significantly inhibited cell migration. *P<0.0001. (D) Representative images from invasion assay. Overexpression of BMP-2 resulted in significantly decreased cell invasion. *P<0.0001. (E) Overexpression of BMP-2 significantly inhibited colony formation ability. *P<0.01.

Mentions: After transient transfection of a plasmid containing human BMP-2 into Caki-1 and Caki-2 cells, significant amounts of BMP-2 protein were detected by Western blotting (Fig. 4A). Cell proliferation (Fig. 4B) and wound healing (Fig. 4C) results demonstrated significant inhibition with BMP-2 transfectants for both Caki-1 and Caki-2 cells as compared to the control vector transfectants. Matrigel invasion assay also showed that the number of invaded cells was significantly decreased in the BMP-2 transfectants as compared with their control counterparts (Fig. 4D). In addition, overexpression of BMP-2 inhibited colony-formation in both Caki-1 and Caki-2 cells (Fig. 4E). Thus, these results suggest that BMP-2 plays an important role in RCC progression.


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)

Effects of BMP-2 overexpression on cell proliferation, migration, invasion, and colony-formation(A) BMP-2 expression level in RCC cell lines (Caki-1, Caki-2) were determined by immunoblotting analysis at 48 hours after transfection of a plasmid containing human BMP-2. (B) Cell viability was analyzed using an MTS cell proliferation assay at 24, 48, 72, and 96 hours after transient transfection. Overexpression of BMP-2 significantly inhibited cell viability. *P<0.05, **P<0.01. (C) Representative images from wound healing assay. After transfection (48 hours), a wound was formed by scraping, then measured 24 hours later. Over-expression of BMP-2 significantly inhibited cell migration. *P<0.0001. (D) Representative images from invasion assay. Overexpression of BMP-2 resulted in significantly decreased cell invasion. *P<0.0001. (E) Overexpression of BMP-2 significantly inhibited colony formation ability. *P<0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Effects of BMP-2 overexpression on cell proliferation, migration, invasion, and colony-formation(A) BMP-2 expression level in RCC cell lines (Caki-1, Caki-2) were determined by immunoblotting analysis at 48 hours after transfection of a plasmid containing human BMP-2. (B) Cell viability was analyzed using an MTS cell proliferation assay at 24, 48, 72, and 96 hours after transient transfection. Overexpression of BMP-2 significantly inhibited cell viability. *P<0.05, **P<0.01. (C) Representative images from wound healing assay. After transfection (48 hours), a wound was formed by scraping, then measured 24 hours later. Over-expression of BMP-2 significantly inhibited cell migration. *P<0.0001. (D) Representative images from invasion assay. Overexpression of BMP-2 resulted in significantly decreased cell invasion. *P<0.0001. (E) Overexpression of BMP-2 significantly inhibited colony formation ability. *P<0.01.
Mentions: After transient transfection of a plasmid containing human BMP-2 into Caki-1 and Caki-2 cells, significant amounts of BMP-2 protein were detected by Western blotting (Fig. 4A). Cell proliferation (Fig. 4B) and wound healing (Fig. 4C) results demonstrated significant inhibition with BMP-2 transfectants for both Caki-1 and Caki-2 cells as compared to the control vector transfectants. Matrigel invasion assay also showed that the number of invaded cells was significantly decreased in the BMP-2 transfectants as compared with their control counterparts (Fig. 4D). In addition, overexpression of BMP-2 inhibited colony-formation in both Caki-1 and Caki-2 cells (Fig. 4E). Thus, these results suggest that BMP-2 plays an important role in RCC progression.

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