BTBD7 promotes the invasion and metastasis of breast cancer MCF-7 cells by targeting Wnt/β-catenin signaling

doi:10.3877/cma. j. issn.1674-0807.2015.06.002

Abstract

Abstract:

Objective

To explore whether knock-down of BTBD7 inhibits the invasion and metastasis of breast cancer cells MCF-7 and the underlying mechanism.

Methods

The expression of BTBD7 was knocked down in MCF-7 cells by lentivirus transfection. The transfected MCF-7 cells served as the experimental group(MCF-7-shBTBD7) and the untransfected cells served as the control group (MCF-7-vec). Then wound healing assay, Transwell assay and MTT method (to draw cell growth curve) were performed to detect the proliferation,invasion and metastasis abilities of MCF-7 cells of two groups. The results were compared between two groups using independent sample t test. Moreover, Cignal finder cancer 10-pathway reporter array was also performed to identify the exact pathway participating in BTBD7 functioning. Western blot assay was performed to confirm the related pathways.

Results

After knocking down BTBD7, wound healing assay showed the migration ability in the experimental group were significantly inhibited compared with the control group[24 h healing rate:(43.38±4.01)% vs (85.93±5.30)%,t=18.108,P＜0.001].Transwell assay demonstrated that the invasion of MCF-7 cells were also significantly suppressed in the experimental group compared with the control group(24 h cell count:50±8 vs 152±7,t=27.378,P＜0.001). MTT assay showed that there was a significant difference between the experimental group and the control group in cell count on day 4 [(3.17±0.23)×10⁴/ml vs(2.58±0.21)×10⁴/ml, t=3.189, P＜0.050], and on day 7 [(7.29±0.37)×10⁴/ml, as compared to the control group(9.57±0.36)×10⁴/ml, t=7.654, P＜0.050]. Result of Cignal finder cancer 10-pathway reporter array by dual luciferase reporter system showed that the relative fluorescence intensity of Wnt signaling pathways was significantly different between two groups(t=12.509, P＜0.001). Western Blot assay confirmed that the protein levels of c-Myc, MMP-7 and β-catenin were significantly different between two groups.

Conclusion

BTBD7 promotes invasion and metastasis of breast cancer MCF-7 cells by targeting Wnt/β-catenin signaling.

Key words: Breast neoplasms; Homeodomain proteins; Wnt proteins; Neoplasm invasiveness;Neoplasm metastasis

Jun Li, Gang Li, Yulian Xie, Minhong Wang. BTBD7 promotes the invasion and metastasis of breast cancer MCF-7 cells by targeting Wnt/β-catenin signaling[J]. Chinese Journal of Breast Disease(Electronic Edition), 2015, 09(06): 357-362.

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组别	Wnt	Notch	p53/DNA damage	TGF-β	Cell cycle/pRb-E2F	NF-κB	Myc/Max	Hypoxia	MAPK/ERK	MAPK/JNK
MCF-7-vec	1	1	1	1	1	1	1	1	1	1
MCF-7-shBTBD7	0.35±0.09	1.02±0.17	0.97±0.17	1.08±0.11	1.14±0.10	1.20±0.13	0.84±0.15	0.71±0.20	0.73±0.15	1.03±0.10
t值	12.509	-0.243	0.300	-1.334	-2.414	-2.667	1.773	2.539	3.003	-0.565
P值	＜0.001	0.831	0.793	0.314	0.137	0.116	0.218	0.126	0.095	0.629

组别	Wnt	Notch	p53/DNA damage	TGF-β	Cell cycle/pRb-E2F	NF-κB	Myc/Max	Hypoxia	MAPK/ERK	MAPK/JNK
MCF-7-vec	1	1	1	1	1	1	1	1	1	1
MCF-7-shBTBD7	0.35±0.09	1.02±0.17	0.97±0.17	1.08±0.11	1.14±0.10	1.20±0.13	0.84±0.15	0.71±0.20	0.73±0.15	1.03±0.10
t值	12.509	-0.243	0.300	-1.334	-2.414	-2.667	1.773	2.539	3.003	-0.565
P值	＜0.001	0.831	0.793	0.314	0.137	0.116	0.218	0.126	0.095	0.629

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