2015; 6:e2007. glucose metabolism from OXPHS to glycolysis in a HIF1- and HIF2-dependent manner. Materials and methods: We obtained data from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) to analyse MTFR2 expression in BC. The prognostic value of MTFR2 expression was assessed using the Kaplan-Meier method. The biological influence of MTFR2 on BC cell lines was studied using proliferation, Transwell migration, invasion and mitochondrial function assays. ValueLow (393)High (607)Age, years 504901663240.00150510227283Tumour Size, mm 205231983250.35420477195282Tumour grade1, 26472354120.0093353158195Venous involvementNegative9143595550.963Positive863452Lymph node metastasisNegative4321502820.010Positive568243325ERNegative253961570.610Positive747297450PRNegative4111662450.556Positive589227362HER2Negative618261357Positive3821322500.016 Open in a separate window Abbreviations: ER, oestrogen receptor; PR, progesterone receptor; HER2, human epidermal growth factor Beta Carotene receptor 2. Kaplan-Meier analysis was used to examine the prognostic value of MTFR2 expression. The results indicated that BC patients with higher MTFR2 expression levels had lower overall survival (OS) rates than those with low MTFR2 levels (ValueHR95% CIValueMTFR2 expression (High vs. Low)1.821.45?2.27 0.011.961.55?2.480.03Age (50 vs. 50)2.241.87?3.330.07Tumour Size ( 20 vs. 20)2.781.24?3.030.26Venous involvement (negative vs. positive)1.110.77?3.83 0.011.621.25?2.090.39Lymph node metastasis (negative vs. positive)3.183.60?9.87 0.011.911.43?2.560.02ER (negative vs. positive)0.760.21?1.170.09PR (negative vs. positive)1.630.69?4.010.08HER2 (negative vs. positive)2.761.39?4.86 0.051.800.65?3.040.05 Open in a separate window Abbreviations: ER, oestrogen receptor; PR, progesterone receptor; HER2, human epidermal growth factor receptor-2. MTFR2 promotes the proliferation, migration and invasion of breast cancer cells To uncover the bio-function of MTFR2 in BC cells, we analysed MTFR2 expression in BC cell lines. Except for MCF-7, the cell lines expressed high levels of MTFR2 (Figure 1F). MTFR2 was stably knocked down in the Hs578T and MDA-231 cell lines and overexpressed in the MCF-7 cell line (Figure 2A). Colony formation assays and CCK-8 assays revealed that higher levels of MTFR2 showed higher proliferation rates in breast cancer cell lines (Figure 2B, ?,2C).2C). We next detected the effect of MTFR2 on cell migration and invasion motility. The results revealed that the capability for cell migration and invasion significantly increased in cells with relatively high levels of MTFR2 (Figure 2D). These results suggest that MTFR2 promotes proliferation, migration and invasion in BC cells. Open in a separate window Figure 2 MTFR promotes the proliferation, migration and invasion of BC. (A) Western blot of MTFR2 in the cell line (NC, Negative Control; OV, overexpression; Sh, small hairpin RNA). (B) The CCK-8 assay of different cell lines (Students two one-tailed paired test * p 0.05). (C) The colony formation assay and statistical analysis of different cell lines (Students two one-tailed paired test * p 0.05). (D) The migration and invasion assays of different cell lines (Students two one-tailed paired test * p 0.05). (E) Western blot of EMT markers of different cell lines. MTFR2 promotes the epithelial-mesenchymal transition of BC cells Our study has revealed that Rabbit Polyclonal to MAP2K3 (phospho-Thr222) MTFR2 promotes proliferation, migration and invasion in BC cells. The EMT phenotype is associated with invasion in cancer cells [20]. The results showed that mesenchymal markers such as N-cadherin, Snail, Vimentin and slug decreased, but epithelial markers such as E-cadherin increased in the MTFR2 knockdown cell lines; however, mesenchymal markers increased, but epithelial markers decreased at both the RNA and protein levels in the MTFR2-overexpressing cell line (Figure 2E). These results suggest that MTFR2 promotes the mesenchymal transition of BC. MTFR2 maintains the aerobic glycolysis of BC cells MTFR2 has rarely been studied in tumourigenesis. However, previous evidence showed that MTFR2 Beta Carotene was correlated with mitochondrial Beta Carotene function. In our study, we found that the activities of mitochondrial complexes I, II and III significantly increased in sh-MTFR2 cells (Figure 3A p 0.001), which is consistent with the levels of the Fe-S-containing subunits Ndufs1 (complex I), SdhB (complex II), and Uqcrfs1 (complex III) (Figure 3B p 0.001). Furthermore, other mitochondrial proteins, such as CytC (cytochrome C) and Fech (ferrochelatase), were also increased in MTFR2 knockdown cells. In contrast, we found that the mitochondrial complexes and proteins of Ndufs1, SdhB, Uqcrfs1, CytC and Fech decreased in MTFR2-overexpressing cells (Figure 3B p 0.001). Open in a separate window Beta Carotene Figure 3 MTFR promotes the glycolysis of BC. (A) The relative activities of the CI CII and CIII of different cell lines (Students two one-tailed paired test * p 0.05). (B) Western blot of OXPHOS markers of different cell lines. (C) The relative viability of different cell lines treated with different inhibitors (Students two one-tailed paired test * p 0.05). (D) The relative ATP level of different cell lines (Students two one-tailed paired test * p 0.05). (E) Western blot of glycolysis markers of different cell lines. (F) The relative lactic acid level of different cell lines (Students.