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Effects of Down-regulating BMP9 on Proliferation and Migration of Human Breast Cancer SK-BR-3 Cells and Its Possible Mechanism in Simulated Bone Microenvironment in vitro
Chen Yingying, Wang Wei, Liu Yuehong, Wan Shaoheng, Zhang Zhihui, Wang Ting, Wang Jinshu, Zhang Yan*
The Key Laboratory of Laboratory Medical Diagnostics in the Ministry of Education, Chongqing Medical University,Chongqing 400016, China
Abstract: To investigate the effects of siRNA-mediated down-regulation of human bone morphogenetic protein 9 (BMP9) on proliferation and migration of human breast cancer SK-BR-3 cells and its possible mechanism in simulated bone microenvironment in vitro， SK-BR-3 cells as blank group， SK-BR-3/RFP cells infected with adenovirus RFP as control group， and SK-BR-3/siBMP9 cells infected with adenovirus siBMP9 as experimental group were indirectly co-cultured with human bone marrow stromal cells HS-5 with Transwell chamber. Effects of downregulating BMP9 on SK-BR-3 cell proliferation and migration were investigated by MTT， wound-healing test and transwell migration test; The expression of related factors were screened by RT-PCR and Western blot. The result showed that in the co-culture system， the expression of BMP9 was decreased in SK-BR-3/siBMP9 group (P<0.05);Down-regulating BMP9 could promote the proliferation of SK-BR-3/siBMP9 cells (P<0.05)， and the wound-healing rate and the number of migratory cells were remarkably increased (P<0.05); Compared with the control group，the expressions of VEGF and CTGF were significantly up-regulated at mRNA and protein levels in SK-BR-3/siBMP9 group (P<0.05)， and Western blot showed p-Akt was higher than the control group (P<0.05). All together，down-regulating BMP9 can promote the proliferation and migration of breast cancer cells SK-BR-3 in microenvironment of bone metastasis， which may be related to up-regulating the expressions of VEGF and CTGF， and PI3K/Akt pathway may be involved in this process.
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