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PHD : Parasites, Hosts and Diseases

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Original Article

Exosomes Secreted by Toxoplasma gondii-Infected L6 Cells: Their Effects on Host Cell Proliferation and Cell Cycle Changes

The Korean Journal of Parasitology 2016;54(2):147-154.
Published online: April 30, 2016

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*Corresponding author (cjy@snu.ac.kr)
• Received: March 19, 2016   • Revised: March 26, 2016   • Accepted: March 27, 2016

© 2016, Korean Society for Parasitology and Tropical Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Exosomes Secreted by Toxoplasma gondii-Infected L6 Cells: Their Effects on Host Cell Proliferation and Cell Cycle Changes
Korean J Parasitol. 2016;54(2):147-154.   Published online April 30, 2016
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Exosomes Secreted by Toxoplasma gondii-Infected L6 Cells: Their Effects on Host Cell Proliferation and Cell Cycle Changes
Korean J Parasitol. 2016;54(2):147-154.   Published online April 30, 2016
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Exosomes Secreted by Toxoplasma gondii-Infected L6 Cells: Their Effects on Host Cell Proliferation and Cell Cycle Changes
Image Image Image Image Image
Fig. 1. (A) Cell growth patterns of uninfected and T. gondii-infected L6 cells. The cell numbers differed significantly at 48 hr and 120 hr post infection (*P<0.05). (B, C) Phase-contrast images of T. gondii-infected cells at 24 hr and 48 hr post infection (MOI=20), respectively. The images show continuous proliferation of T. gondii in host cells. Bar=10 μm. (D) Cell cycle changes induced by T. gondii infection at 24 hr and 48 hr post infection.
Fig. 2. Cell growth patterns of L6 cells grown with usual culture media, T. gondii-infected, treated with 100 μg/ml of exosomes from uninfected (L6 exosome) and T. gondii-infected (RH exosome) cells. The number of L6 cells treated with RH exosome was significantly different from that of the cells treated with L6 exosome (*P=0.001) or that of the cells grown with usual culture media (**P<0.001) at 36 hr by independent t-test.
Fig. 3. Changes of the percentage (%) of cells in the S phase at 12, 24, 36, and 48 hr post T. gondii infection or exosome treatment. Only minute difference is observed between cells treated with L6 exosome or RH tachyzoite exosome at 12 hr post treatment (*P=0.028).
Fig. 4. Changes of the percentage (%) of cells in the G2/M phase at 12, 24, 36, and 48 hr post T. gondii infection or exosome treatment. Cells treated with RH tachyzoite exosome showed decrease in G2/M phase compared to those treated with L6 exosome at 12 hr post treatment (*P=0.030).
Fig. 5. Significantly changed miRNAs in T. gondii RH exosomes compared to those in L6 cell exosomes and their altered signal intensities. Among these 64 miRNAs, those which are specifically associated with cell cycle or cell proliferation are shown in Table 1.
Exosomes Secreted by Toxoplasma gondii-Infected L6 Cells: Their Effects on Host Cell Proliferation and Cell Cycle Changes
miRNA Changes Gene ontology from GOBPa P-value Associated genes
rno-miR-3075 Up-regulated G2/M transition of mitotic cell cycle 4.80E-02 Khdrbs1 Ppm1d
rno-miR-92a-2-5p Up-regulated Regulation of cell proliferation 3.30E-04 Cebpa Fbxo2 Ihh Acvrl1 Calr Carm1 Ccnd2 Hnf4a Id3 Irs1 Itga2 Ngfr Nf1 Nog Nr3c1 Nod2 Nacc1 Pemt Prkca Rac2 Rarg Bcl2l1 S1pr1 Tp53 Vegfa Vash1 Zmiz1
rno-miR-1224 Up-regulated Regulation of cell proliferation 4.70E-02 Ndfip1 Acvrl1 Becn1 Camk2d Rb1 Thrb Vash2
rno-miR-223-3p Up-regulated Regulation of cell proliferation 7.10E-03 Pds5b Rerg F3 Il6st Nfib Rps6kb1 S1pr1 Trim35 Ube2a
rno-miR-325-3p Up-regulated mitotic cell cycle 4.70E-02 Khdrbs1 Pds5b Cdca8 Ccng1 Foxo4 Ppm1d Slfn1 Psmc4 Usp16
rno-miR-200a-3p Down-regulated Cell cycle 2.30E-03 Fbxo5 Pds5b Ruvbl1 Specc1l Apbb2 Cep120 Cyp26b1
Clasp2 Myh10 Psmd11 Ptp4a1 Siah1a Txnip Tgfb2
rno-miR-3072 Down-regulated M phase 4.30E-02 Cep120 Cyp26b1 Lzts2
rno-miR-130b-5p Down-regulated Regulation of growth 2.60E-02 Adam10 Pou3f2 Igfbp2 Lep Ptpn11
Cell cycle 4.20E-02 Cep55 Ccng1 Cyp26b1 Lats2 Mtus1 Ppp6c Rbl2 Txnip Ube2b
rno-miR-3586-5p Down-regulated Cell cycle 1.30E-02 Rassf1 Rassf2 Cyp26b1 Gas2 Mnat1 Mapk13 Nusap1
Pafah1b1 Ppp1cc Ptp4a1 Txnip Tp73
rno-miR-493-3p Down-regulated Cell division 2.90E-02 Clf1 Kif3b Ralbp1
rno-miR-217-3p Down-regulated Regulation of cell proliferation 2.80E-02 Hipk1 Il6st Nr3c1 Nod2 Rarb Tacr1
Table 1. Eleven miRNAs with relevant gene ontology terms

Gene ontology of biological process.