The online version of this article (doi:10.1186/s12885-017-3184-x) contains supplementary material, which is available to authorized users.
Renal cell carcinomas (RCCs) overexpress fatty acid binding protein 7 (FABP7). We chose to study the TUHR14TKB cell line, because it expresses higher levels of FABP7 than other cell lines derived from renal carcinomas (OS-RC-2, 786-O, 769-P, Caki-1, and ACHN).
FABP7 expression was detected using western blotting and real-time PCR. Cell proliferation was determined using an MTS assay and by directly by counting cells. The cell cycle was assayed using flow cytometry. Cell migration was assayed using wound-healing assays. An FABP7 expression vector was used to transfect RCC cell lines.
The levels of FABP7 expressed by TUHR14TKB cells and their doubling times decreased during passage. High-passage TUHR14TKB cells comprised fewer G0/G1-phase and more S-phase cells than low-passage cells. Cell proliferation differed among subclones isolated from cultures of low-passage TUHR14TKB cells. The proliferation of TUHR14TKB cells decreased when FABP7 was overexpressed, and the cell migration property of TUHR14TKB cells were decreased when FABP7 was overexpressed. High concentrations of docosatetraenoic acid and eicosapentaenoic acid accumulated in TUHR14TKB cells that overexpressed FABP7, and docosatetraenoic acid enhanced cell proliferation.
The TUHR14TKB cell line represents a heterogeneous population that does not express FABP7 when it rapidly proliferates. The differences in FABP7 function between RCC cell lines suggests that FABP7 affects cell proliferation depending on cell phenotype.
Additional file 1: Figure S1. Effect of FABP7 overexpression on the 786-O cell line. 786-O cells were cultured for two days in RPMI 1640 medium containing 10% FBS, 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate. a, Western blot analysis of FABP7 expression by cells transfected with the FABP7 vector or control (lacZ) vector. b, Real-time PCR analysis of FABP7 expression of cells transfected with the FABP7 vector or lacZ vector. c-d, The 786-O transfectants were cultured in RPMI-1640 medium containing 10% FBS or 1% FBS with 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate and subjected to cell proliferation and migration assays. c, The doubling times of cell transfected with the FABP7- or the lacZ-expression vector were determined using an MTS assay. The data represent the average and standard deviation (error bars) of five experiments. d, The migration of 786-O cells transfected with the FABP7- or lacZ-expression vector was determined using a wound-healing assay. The data represent the average and standard deviation (error bars) of four experiments. (TIFF 2702 kb)12885_2017_3184_MOESM1_ESM.tiff
Additional file 2: Figure S2. Proliferation of TUHR14TKB cells transfected with an FABP7 expression vector. The proliferation of cells transfected with the FABP7 expression vector or lacZ expression vector was determined using an MTS assay. The data represent of five experiments. Transfectants were cultured in RPMI 1640 medium containing 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate with 1% FBS (a-b) or 10% FBS (c-d). a, c, TUHR14TKB lacZ. b, d, TUHR14TKB FABP7. (TIFF 1521 kb)12885_2017_3184_MOESM2_ESM.tiff
Additional file 3: Table S1. Fatty acid concentrations of TUHR14TKB transfectants. The concentrations of fatty acids accumulated by TUHR14TKB cells transfected with vectors expressing FABP7 or lacZ were assayed in four independent cultures. (XLSX 46 kb)12885_2017_3184_MOESM3_ESM.xlsx
Feng JY, Diao XW, Fan MQ, Wang PX, Xiao Y, Zhong X, et al. Screening of feature genes of the renal cell carcinoma with DNA microarray. Eur Rev Med Pharmacol Sci. 2013;17(22):2994–3001. PubMed
Shi YE, Ni J, Xiao G, Liu YE, Fuchs A, Yu G, et al. Antitumor activity of the novel human breast cancer growth inhibitor, mammary-derived growth inhibitor-related gene. MRG Cancer Res. 1997;57(15):3084–91. PubMed
Wang M, Liu YE, Ni J, Aygun B, Goldberg ID, Shi YE. Induction of Mammary Differentiation by Mammary-derived Growth Inhibitor- related Gene That Interacts with an omega-3 Fatty Acid on Growth Inhibition of Breast Cancer Cells. Cancer Res. 2000;60(22):6482–7. PubMed
RIKEN BioResource Center: Pattern of STR-PCR of TUHR14TKB. http://www2.brc.riken.jp/lab/cell/str_start.shtml?cell_no=RCB1383. Accessed 15 July 2016.
Balendiran GK, Schnutgen F, Scapin G, Borchers T, Xhong N, Lim K, et al. Crystal structure and thermodynamic analysis of human brain fatty acid-binding protein. J Biol Chem. 2000;275(35):27045–54. PubMed
DMC CorpDMC Corp. http://dmed.co.jp. Accessed 15 July 2016
- Functional analysis of fatty acid binding protein 7 and its effect on fatty acid of renal cell carcinoma cell lines
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