BMP9 inhibits the proliferation and invasiveness of breast cancer cells MDA-MB-231 - PubMed (original) (raw)

BMP9 inhibits the proliferation and invasiveness of breast cancer cells MDA-MB-231

Ke Wang et al. J Cancer Res Clin Oncol. 2011 Nov.

Abstract

Background: Transforming growth factor-β (TGF-β) is known to promote tumor proliferation, migration, invasion, and metastasis. Bone morphogenetic proteins (BMPs) are members of the TGF-β superfamily. Several BMPs (BMP2 and BMP7) can enhance the invasion and bone metastasis of breast cancer cells. The function of BMP9, the latest discovered and most powerful osteogenetic factor, in breast cancer has not been fully elucidated.

Methods: BMP9 expression in twenty-three breast cancer patients and three breast cancer cell line types was detected by reverse transcriptase polymerase chain reaction. Changes in proliferation, apoptosis, invasion, and migration in the recombinant MDA-MB-231/BMP9 cells were detected using various assays. The assays were MTT, flow cytometry, colony forming, cell wounding, and transwell invasion. Proliferating cell nuclear antigen and terminal deoxynucleotidy transferase biotin-dUTP nick end labeling staining methods were conducted to detect whether BMP9 affected proliferation and apoptosis in xenogenic mouse models.

Results: Twenty-one of the twenty-three breast cancer patients had amplified BMP9 mRNA transcripts in adjacent non-tumor tissues, although BMP9 was observed in the breast cancer tissue of two patients, its expression was higher in the adjacent non-tumor tissues. BMP9 overexpression inhibited the proliferation, migration, and invasion, as well as induced the apoptosis of the breast cancer cell line MDA-MB-231 in vitro. BMP9 also inhibited tumor growth and induced apoptosis significantly in the xenogenic mouse models.

Conclusions: Decreased BMP9 expression is associated with the elevated proliferation and migration of human breast cancer. BMP9 can inhibit the growth, invasion, and migration of breast cancer cells in vitro and in vivo. BMP9 is a putative tumor suppressor in breast cancer.

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Conflict of interest statement

I declared no Conflict of Interest Statement.

Figures

Fig. 1

BMP9 mRNA expression pattern in breast cancer cell lines. a and b BMP9 mRNA can be detected in HBL-100 and MCF-7 cells, but not in MDA-MB-231 cells (lane 1: HBL-100; lane 2: MCF-7; and lane 3: MDA-MB-231)

Fig. 2

Established recombinant MDA-MB-231/BMP9 that overexpressed BMP9. a Infection efficiency of MDA-MB-231 cells infected with recombinant BMP9 adenovirus for 24 h observed under a fluorescence microscope. b BMP9 mRNA expression pattern in MDA-MB-231/BMP9 cells. BMP9 expression in MDA-MB-231/BMP9 significantly increased after the infection, and _BMP_-9 transcripts increased in the MDA-MB-231/BMP9 cells, compared with the MDA-MB-231/GFP and MDA-MB-231 cells (lane1: MDA-MB-231/BMP9; lane 2: MDA-MB-231/GFP; and lane 3: MDA-MB-231). c BMP9 protein expression pattern in MDA-MB-231/BMP9 cells. Protein level increased in MDA-MB-231/BMP9 cells. The 50 kDa band is the BMP9 protein. d BMP9 protein level in MDA-MB-231/BMP9 cells was also detected using immunocytochemistry staining (×150)

Fig. 3

Decreased growth, migration, and invasiveness of recombinant MDA-MB-231/BMP9 cells caused by BMP9 overexpression. a The effects of BMP9 overexpression on the proliferation of MDA-MB-231. BMP9 overexpression decreased the proliferation of MDA-MB-231 cells (* P < 0.05). b and c The cell cycle distribution in MDA-MB-231/BMP9 cells. BMP9 overexpression blocked the cells at the G2/M phase (* P < 0.05). d The apoptosis percentage increased in MDA-MB-231/BMP9 cells (* P < 0.05). e and f The colony-forming assay results from the various groups. The colony-forming unit decreased significantly in MDA-MB-231/BMP9 cells (* P < 0.05). g and h The wound-closure assay results from the various groups. The wound-closure rate apparently decreased in MDA-MB-231/BMP9 cells (* P < 0.05). i and j The Transwell invasion assay results from the various groups. The amount of invasive cells notably decreased in MDA-MB_231/BMP9 cells (* P < 0.05)

Fig. 3

Fig. 4

Inhibition of tumor formation in vivo caused by BMP9 overexpression. a The tumor sizes in the various groups. b The tumor growth curves of the various groups. The proliferation of MDA-MB-231 cells was inhibited by BMP9 (*P < 0.001). c The hematoxylin-eosin stain of the various groups (×400). The cellular morphology showed no variation among the different groups. d The PCNA stain of the various groups using immunochemical staining. The PCNA-positive cells rate decreased in MDA-MB-231/BMP9 cells (×400) (*P < 0.001). e The TUNEL stain of the various groups. The apoptotic index increased in MDA-MB-231/BMP9 cells (×400) (*P < 0.001)

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BMP9 inhibits the proliferation and invasiveness of breast cancer cells MDA-MB-231 - PubMed (original) (raw)