Abstract
Background
Carbohydrate sulfotransferases 11–13 (CHST11–13), that catalyze the transfer of sulfate to position 4 of the GalNAc residue of chondroitin, have been implicated in various diseases.
Aim
This study aimed to clarify the association of CHST11–13 expression with metastasis and drug sensitivity in hepatocellular carcinoma (HCC) cells.
Methods
We measured the levels of CHST11 and CHST13 in a series of HCC cells using real-time PCR and Western blotting. After RNAi and forced expression treatment of CHST11 and CHST13 in MHCC97L and MHCC97H cells, metastatic potential and drug sensitivity of the two cells were investigated with ECM invasion assay, drug sensitivity assay, and in vivo antitumor activity assay. By real-time PCR and Western blotting, we explored the possible impacts of these two genes on mitogen-activated protein kinase (MAPK) signal pathway. MAPK pathway was blocked by PD98059 or SP600125 to elucidate the effects of MAPK pathway on metastasis and chemosensitivity.
Results
Significantly reduced levels of CHST11 and CHST13 were observed in highly invasive MHCC97H cells compared with those of MHCC97L cell line with low metastatic potential. Decreased or forced expression of CHST11 and CHST13 altered metastatic potential and drug sensitivity of MHCC97L and MHCC97H cells. Remarkable alteration of MAPK activity was shown in two HCC cells with genetic manipulation. Conversely, pharmacologic inhibition of the MAPK pathway suppressed invasive potential and rescued drug sensitivity of MHCC97H cells.
Conclusions
Our results have demonstrated that CHST11 and CHST13 negatively modulate metastasis and drug resistance of HCC cells probably via oncogenic MAPK signal pathway.
Abbreviations
- CHST11–13:
-
Carbohydrate sulfotransferases 11–13 (CHST11–13)
- HCC:
-
Hepatocellular carcinoma
- CS:
-
Chondroitin sulfate
- GAG:
-
Glycosaminoglycan
- C4ST1, 2, 3:
-
Chondroitin-4-O-sulfotransferase-1, 2, 3
- GalNAc:
-
N-acetylgalactosamine
- MAPK:
-
Mitogen-activated protein kinase
- ERK:
-
Extracellular signal-related kinases
- JNK:
-
JunNH2-terminal kinases
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- FBS:
-
Fetal bovine serum
- PCR:
-
Polymerase chain reaction
- shRNA:
-
Short hairpin RNA
- MMP-2:
-
Matrix metalloproteinase 2
References
Bruix J, Sherman M. Management of hepatocellular carcinoma. Hepatology. 2005;42:1208–1236.
Martin TA, Jiang WG. Loss of tight junction barrier function and its role in cancer metastasis. Biochim Biophys Acta. 2009;1788:872–891.
An HJ, Kronewitter SR, de Leoz ML, et al. Glycomics and disease markers. Curr Opin Chem Biol. 2009;13:601–607.
Trerotola M, Ganguly KK, Fazli L, et al. Trop-2 is up-regulated in invasive prostate cancer and displaces FAK from focal contacts. Oncotarget. 2015;6:14318–14328.
Gao Y, Chachadi VB, Cheng PW, et al. Glycosylation potential of human prostate cancer cell lines. Glycoconj J. 2012;29:525–537.
Sugahara K, Kitagawa H. Recent advances in the study of the biosynthesis and functions of sulfated glycosaminoglycans. Curr Opin Struct Biol. 2000;10:518–527.
Klüppel M. The roles of chondroitin-4-sulfotransferase-1 in development and disease. Prog Mol Biol Transl Sci. 2010;93:113–132.
Hiraoka N, Nakagawa H, Ong E, et al. Molecular cloning and expression of two distinct human chondroitin 4-O-sulfotransferases that belong to the HNK-1 sulfotransferase gene family. J Biol Chem. 2000;275:20188–20196.
Okuda T, Mita S, Yamauchi S, et al. Molecular cloning, expression, and chromosomal mapping of human chondroitin 4-sulfotransferase, whose expression pattern in human tissues is different from that of chondroitin 6-sulfotransferase. J Biochem. 2000;128:763–770.
Mikami T, Mizumoto S, Kago N, et al. Specificities of three distinct human chondroitin/dermatan N-acetylgalactosamine 4-O-sulfotransferases demonstrated using partially desulfated dermatan sulfate as an acceptor: implication of differential roles in dermatan sulfate biosynthesis. J Biol Chem. 2003;278:36115–36127.
Kang HG, Evers MR, Xia G, et al. Molecular cloning and characterization of chondroitin-4-O-sulfotransferase-3. A novel member of the HNK-1 family of sulfotransferases. J Biol Chem. 2002;277:34766–34772.
Ricciardelli C, Sakko AJ, Stahl J, et al. Prostatic chondroitin sulfate is increased in patients with metastatic disease but does not predict survival outcome. Prostate. 2009;69:761–769.
Sakko AJ, Butler MS, Byers S, et al. Immunohistochemical level of unsulfated chondroitin disaccharides in the cancer stroma is an independent predictor of prostate cancer relapse. Cancer Epidemiol Biomark Prev. 2008;17:2488–2497.
Herman D, Leakey TI, Behrens A, et al. CHST11 gene expression and DNA methylation in breast cancer. Int J Oncol. 2015;46:1243–1251.
Teng YH, Tan PH, Chia SJ, et al. Increased expression of non-sulfated chondroitin correlates with adverse clinicopathological parameters in prostate cancer. Mod Pathol. 2008;21:893–901.
Cooney CA, Jousheghany F, Yao-Borengasser A, et al. Chondroitin sulfates play a major role in breast cancer metastasis: a role for CSPG4 and CHST11 gene expression in forming surface P-selectin ligands in aggressive breast cancer cells. Breast Cancer Res. 2011;13:R58. doi:10.1186/bcr2895.
Schmidt HH, Dyomin VG, Palanisamy N, et al. Deregulation of the carbohydrate (chondroitin 4) sulfotransferase 11 (CHST11) gene in a B-cell chronic lymphocytic leukemia with a t(12;14)(q23;q32). Oncogene. 2004;23:6991–6996.
Basappa, Murugan S, Sugahara KN, et al. Involvement of chondroitin sulfate E in the liver tumor focal formation of murine osteosarcomacells. Glycobiology. 2009;19:735–742.
Koul HK, Pal M, Koul S. Role of p38 map kinase signal transduction in solid tumors. Genes Cancer. 2013;4:342–359.
Xie J, Jin B, Li DW, et al. ABCG2 regulated by MAPK pathways is associated with cancer progression in laryngeal squamous cell carcinoma. Am J Cancer Res. 2014;4:698–709.
Zuo L, Lu M, Zhou Q, et al. Butyrate suppresses proliferation and migration of RKO colon cancer cells though regulating endocan expression by MAPK signaling pathway. Food Chem Toxicol. 2013;62:892–900.
Hsieh YH, Wu TT, Huang CY, et al. p38 mitogen-activated protein kinase pathway is involved in protein kinase Calpha-regulated invasion in human hepatocellular carcinoma cells. Cancer Res. 2007;67:4320–4327.
Yang L, Ling Y, Zhang Z, et al. ZL11n is a novel nitric oxide-releasing derivative of farnesylthiosalicylic acid that induces apoptosis in human hepatoma HepG2 cells via MAPK/mitochondrial pathways. Biochem Biophys Res Commun. 2011;409:752–757.
Klüppel M, Samavarchi-Tehrani P, Liu K, et al. C4ST-1/CHST11-controlled chondroitin sulfation interferes with oncogenic HRAS signaling in Costello syndrome. Eur J Hum Genet. 2012;20:870–877.
Li Y, Tang ZY, Ye SL, et al. Establishment of cell clones with different metastatic potential from the metastatic hepato-cellular carcinoma cell line MHCC97. World J Gastroenterol. 2001;7:630–636.
Tian J, Tang ZY, Ye SL, et al. New human hepatocellular carcinoma (HCC) cell line with highly metastatic potential (MHCC97) and its expressions of the factors associated with metastasis. Br J Cancer. 1999;81:814–821.
Schütte K, Bornschein J, Malfertheiner P. Hepatocellular carcinoma–epidemiological trends and risk factors. Dig Dis. 2009;27:80–92.
Gorelik E, Galili U, Raz A. On the role of cell surface carbohydrates and their binding proteins (lectins) in tumor metastasis. Cancer Metastasis Rev. 2001;20:245–277.
Basappa, Rangappa KS, Sugahara K. Roles of glycosaminoglycans and glycanmimetics in tumor progression and metastasis. Glycoconj J. 2014;31:461–467.
Vynios DH, Theocharis DA, Papageorgakopoulou N, et al. Biochemical changes of extracellular proteoglycans in squamous cell laryngeal carcinoma. Connect Tissue Res. 2008;49:239–243.
Stylianou M, Skandalis SS, Papadas TA, et al. Stage-related decorin and versican expression in human laryngeal cancer. Anticancer Res. 2008;28:245–251.
Honke K, Taniguchi N. Sulfotransferases and sulfated oligosaccharides. Med Res Rev. 2002;22:637–654.
Klüppel M, Wight TN, Chan C, et al. Maintenance of chondroitin sulfation balance by chondroitin-4-sulfotransferase 1 is required for chondrocyte development and growth factor signaling during cartilage morphogenesis. Development. 2005;132:3989–4003.
Bergefall K, Trybala E, Johansson M, et al. Chondroitin sulfate characterized by the E-disaccharide unit is a potent inhibitor of herpes simplex virus infectivity and provides the virus binding sites on gro2C cells. J Biol Chem. 2005;280:32193–32199.
Karlsson C, Dehne T, Lindahl A, et al. Genome-wide expression profiling reveals new candidate genes associated with osteoarthritis. Osteoarthr Cartil. 2010;18:581–592.
ten Dam GB, van de Westerlo EM, Purushothaman A, et al. Antibody GD3G7 selected against embryonic glycosaminoglycans defines chondroitin sulfate-E domains highly up-regulated in ovarian cancer and involved in vascular endothelial growth factor binding. Am J Pathol. 2007;171:1324–1333.
Bret C, Hose D, Reme T, et al. Expression of genes encoding for proteins involved in heparan sulphate and chondroitin sulphate chain synthesis and modification in normal and malignant plasma cells. Br J Haematol. 2009;145:350–368.
Kalathas D, Theocharis DA, Bounias D, et al. Alterations of glycosaminoglycan disaccharide content and composition in colorectal cancer: structural and expressional studies. Oncol Rep. 2009;22:369–375.
Wegrowski Y, Maquart FX. Chondroitin sulfate proteoglycans in tumor progression. Adv Pharmacol. 2006;53:297–321.
Asada M, Shinomiya M, Suzuki M, et al. Glycosaminoglycan affinity of the complete fibroblast growth factor family. Biochim Biophys Acta. 2009;1790:40–48.
Deepa SS, Umehara Y, Higashiyama S, et al. Specific molecular interactions of oversulfated chondroitin sulfate E with various heparin-binding growth factors. Implications as a physiological binding partner in the brain and other tissues. J Biol Chem. 2002;277:43707–43716.
Iida J, Wilhelmson KL, Ng J, et al. Cell surface chondroitin sulfate glycosaminoglycan in melanoma: role in the activation of pro-MMP-2 (pro-gelatinase A). Biochem J. 2007;403:553–563.
Willis CM, Wrana JL, Klüppel M. Identification and characterization of TGFbeta-dependent and -independent cis-regulatory modules in the C4ST-1/CHST11 locus. Genet Mol Res. 2009;8:1331–1343.
Nadanaka S, Ishida M, Ikegami M, et al. Chondroitin 4-O-sulfotransferase-1 modulates Wnt-3a signaling through control of E disaccharide expression of chondroitin sulfate. J Biol Chem. 2008;283:27333–27343.
Shortkroff S, Yates KE. Alteration of matrix glycosaminoglycans diminishes articular chondrocytes’ response to a canonical Wnt signal. Osteoarthr Cartil. 2007;15:147–154.
Cheng SB, Wu LC, Hsieh YC, et al. Supercritical carbon dioxide extraction of aromatic turmerone from Curcuma longa Linn. Induces apoptosis through reactive oxygen species-triggered intrinsic and extrinsic pathways in human hepatocellular carcinoma HepG2 cells. J Agric Food Chem. 2012;60:9620–9630.
Acknowledgments
This work was supported by Grants from National Natural Science Foundation of China (81271910, 81472014) and from Natural Science Foundation of Liaoning Province, China (2014023043), and supported by Project for Liaoning BaiQianWan Talents Program (2012921014).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Zhou, H., Li, Y., Song, X. et al. CHST11/13 Regulate the Metastasis and Chemosensitivity of Human Hepatocellular Carcinoma Cells Via Mitogen-Activated Protein Kinase Pathway. Dig Dis Sci 61, 1972–1985 (2016). https://doi.org/10.1007/s10620-016-4114-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10620-016-4114-5