Liu Yang, Zhang Ran-Xi, Yuan Wei, Chen Hou-Qing, Tian Dong-Dong, Li Hao, Jiang Xiao, Deng Zhong-Liang, Wang Yang
Department of Orthopaedics, Second Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Department of Nephrology, Second Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Cell Physiol Biochem. 2018;45(5):1759-1771. doi: 10.1159/000487784. Epub 2018 Feb 23.
BACKGROUND/AIMS: Bone morphogenetic proteins (BMPs) and BMP receptors widely participate in osteolytic metastasis of breast cancer, while their role in tumor-stromal interaction is largely unknown. In this study, we investigated whether BMP receptor type 1a (BMPR1a) can alter the interaction between metastatic cancer cells and osteoclast precursors.
Adenovirus-mediated RNA interference was used to interrupt target genes of human breast cancer cell lines and nude mice were injected intratibially with the cancer cells. Tumor-bearing mice were examined by bioluminescence imaging and microCT. Sections of metastatic legs were measured by a series of staining methods. Murine bone marrow mononuclear cells or RAW264.7 cells were cultured with conditioned media of breast cancer cells. RT-PCR, Western blotting and ELISA were used to test mRNA and protein expressions of target molecules.
Expression of BMPR1a of MDA-MB-231-luc cells at tumor-bone interface was apparently stronger than that of cancer cells distant from the interface. Mice injected with BMPR1a-knockdown MDA-MB-231-luc cells showed reduced tumor growth and bone destruction compared with control groups. Knockdown (KD) of BMPR1a of MDA-MB-231-luc cells or MCF-7 cells decreased the level of receptor activator for NF-κB ligand (RANKL). Level of RANKL in MDA-MB-231-luc cells or MCF-7 cells was reduced by p38 inhibitor. Compared with control group, knockdown of p38 of breast cancer cells decreased cancer-induced osteoclastogenesis.
Knockdown of BMPR1a of breast cancer cells suppresses their production of RANKL via p38 pathway and inhibits cancer-induced osteoclastogenesis, which indicates that BMPR1a might be a possible target in breast cancer-induced osteolytic metastasis.
背景/目的:骨形态发生蛋白(BMPs)和BMP受体广泛参与乳腺癌的溶骨性转移,但其在肿瘤-基质相互作用中的作用尚不清楚。在本研究中,我们调查了1a型BMP受体(BMPR1a)是否能改变转移性癌细胞与破骨细胞前体之间的相互作用。
采用腺病毒介导的RNA干扰技术干扰人乳腺癌细胞系的靶基因,并将癌细胞经胫骨内注射到裸鼠体内。通过生物发光成像和显微CT对荷瘤小鼠进行检查。用一系列染色方法测量转移腿部的切片。将小鼠骨髓单核细胞或RAW264.7细胞与乳腺癌细胞的条件培养基一起培养。采用RT-PCR、蛋白质印迹法和ELISA检测靶分子的mRNA和蛋白表达。
MDA-MB-231-luc细胞在肿瘤-骨界面处的BMPR1a表达明显强于远离界面的癌细胞。与对照组相比,注射BMPR1a基因敲低的MDA-MB-231-luc细胞的小鼠肿瘤生长和骨破坏减少。MDA-MB-231-luc细胞或MCF-7细胞的BMPR1a基因敲低(KD)降低了核因子κB受体激活蛋白配体(RANKL)的水平。p38抑制剂降低了MDA-MB-231-luc细胞或MCF-7细胞中RANKL的水平。与对照组相比,乳腺癌细胞p38基因敲低减少了癌症诱导的破骨细胞生成。
乳腺癌细胞的BMPR1a基因敲低通过p38途径抑制其RANKL的产生,并抑制癌症诱导的破骨细胞生成,这表明BMPR1a可能是乳腺癌诱导的溶骨性转移的一个潜在靶点。