Integral regulation of CHI3L1 gene and ERVW-1 locus expression by heparin in the gioblastoma cell lines U-87 MG and U-251 MG

  • O. V. Anopriyenko
  • P. O. Areshkov
  • O. V. Zhuk
  • V. A. Shablii
  • I. Ya. Skrypkina

Abstract

Aim. To study the effect of heparin, as a factor of tumor microenvironment, on the expression of ERVW-1 locus and CHI3L1 gene, involved in increasing of cancer cells metastatic potential, in U-87 MG (U87) and U-251 MG (U251) gliotblastoma cells. Methods. U87 and U251 cells were cultured with or without the addition of heparin to the cultural medium. Total cellular RNA was isolated and used for cDNA synthesis and subsequent PCR with primers to different regions of ERVW-1 and CHI3L1. Results. Both U87 and U251 cells showed high level of expression of full-length ERVW-1 RNA. But only in U251 cells inhibition of the full-length transcript by heparin was revealed. Spliced isoform hasn’t been detected in any of variants. Though the solely env gene transcript expressed at low level in both lines and was inhibited similarly by heparin. Expression of CHI3L1 has been detected in both lines with primers towards exons 4-5 with some lowering in level under the heparin influence. Only in U87 cells the PCR-fragment with primers to exons 8-9 has been detected. In control U87 cells hypothetic spliced isoform of CHI3L1 transcript has been revealed with primers to exons 1-9. Conclusions. Heparin has complex and cell line-dependent regulation of expression of ERVW-1 locus and CHI3L1 gene in the glioblastoma cell lines U87 and U251.

Keywords: glioblastoma, heparin, ERVW-1, CHI3L1.

References

Iwadate Y. Plasticity in Glioma Stem Cell Phenotype and Its Therapeutic Implication. Neurol Med Chir (Tokyo). 2018. Vol. 58, № 2. P. 61–70. doi: 10.2176/nmc.ra.2017-0089.

Patil V., Pal J., Somasundaram K. Elucidating the cancer-specific genetic alteration spectrum of glioblastoma derived cell lines from whole exome and RNA sequencing. Oncotarget. 2015. Vol. 6, № 41. P. 43452–43471. doi: 10.18632/oncotarget.6171.

Schnoor R., Maas S.L., Broekman M.L. Heparin in malignant glioma: review of preclinical studies and clinical results. J. Neurooncol. 2015. Vol. 124, № 2. P. 151–156. doi: 10.1007/s11060-015-1826-x.

Attarha S., Roy A., Westermark B., Tchougounova E. Mast cells modulate proliferation, migration and stemness of glioma cells through downregulation of GSK3в expression and inhibition of STAT3 activation. Cell Signal. 2017. Vol. 37. P. 81–92. doi: 10.1016/j.cellsig.2017.06.004.

Ren W., Guo Q., Yang Y., Chen F. bFGF and heparin but not laminin are necessary factors in the mediums that affect NSCs differentiation into cholinergic neurons. Neurol Res. 2006. Vol. 28, № 1. P. 87–90. doi: 10.1179/016164106X91933.

Simard J.M., Tosun C., Ivanova S., Kurland D.B., Hong C., Radecki L., Gisriel C., Mehta R., Schreibman D., Gerzanich V. Heparin reduces neuroinflammation and transsynaptic neuronal apoptosis in a model of subarachnoid hemorrhage. Transl. Stroke Res. 2012. Vol. 3, Suppl. 1. P. 155–165. doi: 10.1007/s12975-012-0166-9.

Areshkov P.O., Avdieiev S.S., Iershov A.V., Kavsan V.M. Stimulation of transient versus sustained ERK1/2 phosphorylation by relative chitinase-like proteins CHI3L1 and CHI3L2 correlates with different kinase localization and biological outcome. Biopolymers and Cell. 2011. P. 343–346. URL: http://dx.doi.org/10.7124/bc.00011F.

Kzhyshkowska J., Yin S., Liu T., Riabov V., Mitrofanova I. Role of chitinase-like proteins in cancer. Biol. Chem. 2016. Vol. 397, № 3. P. 231–247. doi: 10.1515/hsz-2015-0269.

Bjerregaard B., Holck S., Christensen I.J., Larsson L.I. Syncytin is involved inbreast cancer-endothelial cell fusions. Cell. Mol. Life Sci. 2006. Vol. 63, № 3. 16. P. 1906–1911.

Geng B., Pan J., Zhao T., Ji J., Zhang C., Che Y., Yang J., Shi H., Li J., Zhou H., Mu X., Xu C., Wang C., Xu Y., Liu Z., Wen H., You Q. Chitinase 3-like 1-CD44 interaction promotes metastasis and epithelial-to-mesenchymal transition through в-catenin/Erk/Akt signaling in gastric cancer. J. Exp. Clin. Cancer Res. 2018. Vol. 37, № 1. P. 208. doi: 10.1186/s13046-018-0876-2.

Liu C., Xu J., Wen F., Yang F., Li X., Geng D., Li L., Chen J., Zheng J. Upregulation of syncytin-1 promotes invasion and metastasis by activating epithelial-mesenchymal transition-related pathway in endometrial carcinoma. OncoTargets Ther. 2018. Vol. 12. P. 31–40. doi: 10.2147/OTT.S191041.

Beneљovб M., Trejbalovб K., Kovбшovб D., Vernerovб Z., Hron T., Kuиerovб D., Hejnar J. DNA hypomethylation and aberrant expression of the human endogenous retrovirus ERVWE1/syncytin-1 in seminomas. Retrovirology. 2017. Vol. 14, № 1. P. 20. doi: 10.1186/s12977-017-0342-9.

Dнaz-Carballo D., Klein J., Acikelli A.H., Wilk C., Saka S., Jastrow H., Wennemuth G., Dammann P., Giger-Pabst U., Khos-rawipour V., Rassow J., Nienen M., Strumberg D. Cytotoxic stress induces transfer of mitochondria-associated human en-dogenous retroviral RNA and proteins between cancer cells. Oncotarget. 2017. Vol. 8, № 56. P. 95945–95964. doi: 10.18632/oncotarget.21606.

Oh S.J., Yang J.I., Kim O., Ahn E.J., Kang W.D., Lee J.H., Moon K.S., Lee K.H., Cho D. Human U87 glioblastoma cells with stemness features display enhanced sensitivity to natural killer cell cytotoxicity through altered expression of NKG2D ligand. Cancer Cell Int. 2017. Vol. 17. P. 22. doi: 10.1186/s12935-017-0397-7.