The Co-cultural Interaction of Mice's Early Embryo and Tumor Cells <i>in Vitro</i>_West China Medical Journal

Authors：

 JiangShasha ^1,2 , TanChunlu ³ , ZhangHao ³ , LuoFeng ¹ ,  LiKezhou ³

1. Department of Oncology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China;
2. Department of Oncology, the 3rd People's Hospital of Chengdu, Chengdu, Sichuan 610031, P. R. China;
3. Department of Pancreatic Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China;

Corresponding author：

LiKezhou, Email: huaxipancreas@163.com

Keywords：

Mice's early embryo; Tumor cell; Co-culture; Tumor proliferation; Tumor apoptosis

DOI：

10.7507/1002-0179.201600416

Video：

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Objective To establish the co-culture model of mice's early embryo and tumor cells in Vitro to observe the embryonic development and biologic behavior of tumor cells in the same microenvironment and discuss their interaction. Methods We acquired 2-cell embryos from mice, and then co-cultured them with tumor cell lines of mice in Vitro. We observed the development of embryos in Vitro and the rates of 4-cell embryos, morula and blastocyst formation. The transwell chamber was used for culture. Methylthiazolyldiphenyl-tetrazolium method was used to test the proliferative activity of tumor cells, while the flow cytometry was used to test its apoptosis. The interaction of co-cultured embryos and tumor cells was analyzed by propidium iodide staining and immunohistochemical technique. Results The co-cultured 2-cell embryos could continue surviving and developing. The rates of 4-cell embryos, morula and blastocyst formation increased significantly in the co-cultured group (P<0.05). There was no significant difference in the proliferative activity and apoptosis of tumor cells between the co-cultured group and the control group (P>0.05). Tumor-free ring formed between the trophoblast and tumor cells. We could observe tumor cells stacked around the tumor-free ring. However, no difference in expression of proliferating cell nuclear antigen and B-cell lymphoma leukemia-2 was observed in tumor cells stacking around the tumor-free ring compared with those elsewhere. Conclusion The development of 2-cell embryos is enhanced in the co-culture model. The proliferative activity and apoptosis of tumor cells are not affected in this model. A tumor-free ring can form between trophoblast and tumor cells. However, the proliferative activity of tumor cells is not affected by this ring.

Citation： JiangShasha, TanChunlu, ZhangHao, LuoFeng, LiKezhou. The Co-cultural Interaction of Mice's Early Embryo and Tumor Cells in Vitro. West China Medical Journal, 2016, 31(9): 1525-1529. doi: 10.7507/1002-0179.201600416 Copy

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2.	Clevers H, Nusse R. Wnt/β-Catenin signaling and disease[J]. Cell, 2012, 149(6):1192-1205.
3.	Glienke W, Hausmann E, Bergmann L. Downregulation of STAT3 signaling induces apoptosis but also promotes anti-apoptotic gene expression in human pancreatic cancer cell lines[J]. Tumour Biol, 2011, 32(3):493-500.
4.	Nordström L, Andersson E, Kuci V, et al. DNA methylation and histone modifications regulate SOX11 expression in lymphoid and solid cancer cells[J]. BMC Cancer, 2015, 15:273.
5.	Gibadulinova A, Tothova V, Pastorek J, et al. Transcriptional regulation and functional implication of S100P in cancer[J]. Amino Acids, 2011, 41(4):885-892.
6.	Carosella ED, Rouas-Freiss N, Roux DT, et al. HLA-G:an immune checkpoint molecule[J]. Adv Immunol, 2015, 127:33-144.
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8.	Hochedlinger K, Blelloch R, Brennan C, et al. Reprogramming of a melanoma genome by nuclear transplantation[J]. Genes Dev, 2004, 18(15):1875-1885.
9.	Kulesa PM, Kasemeier-Kulesa JC, Teddy JM, et al. Reprogramming metastatic melanoma cells to assume a neural crest cell-like phenotype in an embryonic microenvironment[J]. Proc Natl Acad Sci USA, 2006, 103(10):3752-3757.
10.	Raof NA, Mooney BM, Xie Y. Bioengineering embryonic stem cell microenvironments for the study of breast cancer[J]. Int J Mol Sci, 2011, 12(11):7662-7691.
11.	Tan XW, Ma SF, Yu JN, et al. Effects of species and cellular activity of oviductal epithelial cells on their dialogue with co-cultured mouse embryos[J]. Cell Tissue Res, 2007, 327(1):55-66.
12.	Taniguchi F, Harada T, Nara M, et al. Coculture with a human granulosa cell line enhanced the development of murine preimplantation embryos via SCF/c-kit system[J]. J Assist Reprod Genet, 2004, 21(6):223-228.
13.	Andras N, Marina G, Kristina V, et al. Manipulating the mouse embryo:a laboratory manual[M]. New York:Cold Spring Harbor Laboratory Press, 2006:154-158.
14.	Sell S, Nicolini A, Ferrari P, et al. Cancer:a problem of developmental biology; scientific evidence for reprogramming and differentiation therapy[J]. Curr Drug Targets, 2016, 17(10):1103-1110.
15.	Díez-Torre A, Andrade R, Eguizábal C, et al. Reprogramming of melanoma cells by embryonic microenvironments[J]. Int J Dev Biol, 2009, 53(8/9/10):1563-1568.
16.	Bischof AG, Yüksel D, Mammoto T, et al. Breast cancer normalization induced by embryonic mesenchyme is mediated by extracellular matrix biglycan[J]. Integr Biol (Camb), 2013, 5(8):1045-1056.
17.	Sundaram MV. The love-hate relationship between Ras and Notch[J]. Genes Dev, 2005, 19(16):1825-1839.
18.	Nissi R, Talvensaari-Mattila A, Kotila V, et al. Circulating matrix metalloproteinase MMP-9 and MMP-2/TIMP-2 complex are associated with spontaneous early pregnancy failure[J]. Reprod Biol Endocrinol, 2013, 11(2):2.
19.	Tseng CN, Huang CF, Cho CL, et al. Brefeldin a effectively inhibits cancer stem Cell-Like properties and MMP-9 activity in human colorectal cancer Colo 205 cells[J]. Molecules, 2013, 18(9):10242-10253.
20.	Na KH, Lee HJ, Choi JH, et al. Dynamic alterations in integrin α4 expression by hypoxia are involved in trophoblast invasion during early implantation[J]. J Cell Biochem, 2012, 113(2):685-694.

1. Luo W, Li S, Peng B, et al. Embryonic stem cells markers SOX2, OCT4 and Nanog expression and their correlations with epithelial-mesenchymal transition in nasopharyngeal carcinoma[J]. PLoS One, 2013, 8(2):e56324.
2. Clevers H, Nusse R. Wnt/β-Catenin signaling and disease[J]. Cell, 2012, 149(6):1192-1205.
3. Glienke W, Hausmann E, Bergmann L. Downregulation of STAT3 signaling induces apoptosis but also promotes anti-apoptotic gene expression in human pancreatic cancer cell lines[J]. Tumour Biol, 2011, 32(3):493-500.
4. Nordström L, Andersson E, Kuci V, et al. DNA methylation and histone modifications regulate SOX11 expression in lymphoid and solid cancer cells[J]. BMC Cancer, 2015, 15:273.
5. Gibadulinova A, Tothova V, Pastorek J, et al. Transcriptional regulation and functional implication of S100P in cancer[J]. Amino Acids, 2011, 41(4):885-892.
6. Carosella ED, Rouas-Freiss N, Roux DT, et al. HLA-G:an immune checkpoint molecule[J]. Adv Immunol, 2015, 127:33-144.
7. Bagley RG, Honma N, Weber W, et al. Endosialin/TEM 1/CD248 is a pericyte marker of embryonic and tumor neovascularization[J]. Microvasc Res, 2008, 76(3):180-188.
8. Hochedlinger K, Blelloch R, Brennan C, et al. Reprogramming of a melanoma genome by nuclear transplantation[J]. Genes Dev, 2004, 18(15):1875-1885.
9. Kulesa PM, Kasemeier-Kulesa JC, Teddy JM, et al. Reprogramming metastatic melanoma cells to assume a neural crest cell-like phenotype in an embryonic microenvironment[J]. Proc Natl Acad Sci USA, 2006, 103(10):3752-3757.
10. Raof NA, Mooney BM, Xie Y. Bioengineering embryonic stem cell microenvironments for the study of breast cancer[J]. Int J Mol Sci, 2011, 12(11):7662-7691.
11. Tan XW, Ma SF, Yu JN, et al. Effects of species and cellular activity of oviductal epithelial cells on their dialogue with co-cultured mouse embryos[J]. Cell Tissue Res, 2007, 327(1):55-66.
12. Taniguchi F, Harada T, Nara M, et al. Coculture with a human granulosa cell line enhanced the development of murine preimplantation embryos via SCF/c-kit system[J]. J Assist Reprod Genet, 2004, 21(6):223-228.
13. Andras N, Marina G, Kristina V, et al. Manipulating the mouse embryo:a laboratory manual[M]. New York:Cold Spring Harbor Laboratory Press, 2006:154-158.
14. Sell S, Nicolini A, Ferrari P, et al. Cancer:a problem of developmental biology; scientific evidence for reprogramming and differentiation therapy[J]. Curr Drug Targets, 2016, 17(10):1103-1110.
15. Díez-Torre A, Andrade R, Eguizábal C, et al. Reprogramming of melanoma cells by embryonic microenvironments[J]. Int J Dev Biol, 2009, 53(8/9/10):1563-1568.
16. Bischof AG, Yüksel D, Mammoto T, et al. Breast cancer normalization induced by embryonic mesenchyme is mediated by extracellular matrix biglycan[J]. Integr Biol (Camb), 2013, 5(8):1045-1056.
17. Sundaram MV. The love-hate relationship between Ras and Notch[J]. Genes Dev, 2005, 19(16):1825-1839.
18. Nissi R, Talvensaari-Mattila A, Kotila V, et al. Circulating matrix metalloproteinase MMP-9 and MMP-2/TIMP-2 complex are associated with spontaneous early pregnancy failure[J]. Reprod Biol Endocrinol, 2013, 11(2):2.
19. Tseng CN, Huang CF, Cho CL, et al. Brefeldin a effectively inhibits cancer stem Cell-Like properties and MMP-9 activity in human colorectal cancer Colo 205 cells[J]. Molecules, 2013, 18(9):10242-10253.
20. Na KH, Lee HJ, Choi JH, et al. Dynamic alterations in integrin α4 expression by hypoxia are involved in trophoblast invasion during early implantation[J]. J Cell Biochem, 2012, 113(2):685-694.

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