Secretomes of Primary Cancer-associated Fibroblasts Upregulate the Expression of Stemness Markers in HT-29 Human Colorectal Carcinoma Cells

Septelia Inawati Wanandi, Dwi Retna Lestari, Noza Hilbertina, Nurjati Chairani Siregar, Sri Widia Jusman, Murdani Abdullah

Abstract


BACKGROUND: Cancer-associated fibroblast (CAF) is the most abundant tumor stroma. Our previous study has demonstrated that the secretomes of CAF isolated from colorectal carcinoma (CRC) patients could induce epithelial-mesenchymal transition in the HT-29 CRC cell line. However, the role of CAF secretomes in CRC stemness is needed to be further investigated. Therefore, the present study aimed to investigate the effect of CAF secretomes from CRC patients on the expression of stemness markers in HT-29 CRC cells in comparison with the secretomes from normal fibroblasts.

METHODS: Fibroblasts were isolated from tumor (CAF) and their counterpart non-tumor (NF) areas of three CRC patients undergone surgical resection. Normal preputium fibroblasts (PF) were isolated during circumcision of three healthy boys aged 8 years. All fibroblasts were grown in free-serum culture medium for 24 hours to collect 50% (v/v) conditioned medium (CM). Then, CM was supplemented to HT-29 CRC cells for 72 hours. The effects of CAF- and NF-CM on the mRNA expression of CD44, CD133, OCT4, and ALDH1A1 were analysed using qRT-PCR. Cells proliferation was measured using the trypan blue exclusion assay.

RESULTS: Supplementation of CAF-CM (50% v/v) significantly increased CD44, CD133, OCT4, and ALDH1A1 mRNA expressions compared to that of NF-CM and control without supplementation but had no effect on the proliferation of HT-29 cells.

CONCLUSION: CAF secretomes from CRC patients upregulate the expression of CRC stemness.

KEYWORDS: cancer-associated fibroblasts, ALDH1A1, OCT4, CD44, CD133, colorectal carcinoma


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Zhou JY, Chen M, Ma L, Wang X, Chen YG, Liu SL. Role of CD44high/CD133high HCT-116 cells in the tumorigenesis of colon cancer. Oncotarget. 2016; 7: 7657-66, CrossRef.

Zhao Y, Peng J, Zhang E, Jiang N, Li J, Zhang Q, et al. CD133 expression may be useful as a prognostic indicator in colorectal cancer, a tool for optimizing therapy and supportive evidence for the cancer stem cell hypothesis: a meta-analysis. Oncotarget. 2016; 7: 10023-36, CrossRef.

Michl M, Heinemann V, Jung A, Engel J, Kirchner T, Neumann J. Expression of cancer stem cell markers in metastatic colorectal cancer correlates with liver metastasis, but not with metastasis to the central nervous system. Pathol Res Pract. 2015; 211: 601-9, CrossRef.

Tomita H, Tanaka K, Tanaka T, Hara A. Aldehyde dehydrogenase 1A1 in stem cells and cancer. Oncotarget. 2016; 7: 11018-32, CrossRef.

Liu X, Huang J, Chen T, Wang Y, Xin S, Li J, et al. Yamanaka factors critically regulate the developmental signaling network in mouse embryonic stem cells. Cell Res. 2008; 18: 1177-89, CrossRef.

Garza-Treviño EN, Said-Fernández SL, Martínez-Rodríguez HG. Understanding the colon cancer stem cells and perspectives on treatment. Cancer Cell Int. 2015; 15: 2, CrossRef.

Han XY, Wei B, Fang JF, Zhang S, Zhang FC, Zhang HB, et al. Epithelial-mesenchymal transition associates with maintenance of stemness in spheroid-derived stem-like colon cancer cells. PLoS One. 2013; 8: e73341, CrossRef.

Tommelein J, Verset L, Boterberg T, Demetter P, Bracke M, De Wever O. Cancer-associated fibroblasts connect metastasis-promoting communication in colorectal cancer. Front Oncol. 2015; 5: 1-11, CrossRef.

Das M, Law S. Role of tumor microenvironment in cancer stem cell chemoresistance and recurrence. Int J Biochem Cell Biol. 2018; 103: 115-24, CrossRef.

Wanandi SI, Hilbertina N, Siregar NC, Abdullah M, Jeo WS. CAF secretomes induce epithelial-mesenchymal transition on HT-29 colorectal carcinoma cells associated with HGF signaling. J Pakistan Med Assoc. 2020; in press.

Chen WJ, Ho CC, Chang YL, Chen HY, Lin CA, Ling TY, et al. Cancer-associated fibroblasts regulate the plasticity of lung cancer stemness via paracrine signalling. Nat Commun. 2014; 5: 3472, CrossRef.

Álvarez-Teijeiro S, García-Inclán C, Villaronga MÁ, Casado P, Hermida-Prado F, Granda-Díaz R, et al. Factors secreted by cancer-associated fibroblasts that sustain cancer stem properties in head and neck squamous carcinoma cells as potential therapeutic targets. Cancers (Basel). 2018; 10: 334, CrossRef.

Izumi D, Toden S, Ureta E, Ishimoto T, Baba H, Goel A. TIAM1 promotes chemoresistance and tumor invasiveness in colorectal cancer. Cell Death Dis. 2019; 10: 267, CrossRef.

Ren J, Ding L, Zhang D, Shi G, Xu Q, Shen S, et al. Carcinoma-associated fibroblasts promote the stemness and chemoresistance of colorectal cancer by transferring exosomal lncRNA H19. Theranostics. 2018; 8: 3932-48, CrossRef.

Herrera M, Herrera A, Larriba MJ, Mayorga GF, de Herreros AG. Colon cancer-associated fibroblast establishment and culture growth. 2016; 6: 1773, CrossRef.

Hosea R, Hardiany N, Ohneda O, Wanandi S. Glucosamine decreases the stemness of human ALDH+ breast cancer stem cells by inactivating STAT3. Oncol Lett. 2018; 16: 4737-44, CrossRef.

Wanandi SI, Syahrani RA, Arumsari S, Wideani G, Hardiany NS. Profiling of gene expression associated with stemness and aggressiveness of ALDH1A1-expressing human breast cancer cells. Malaysian J Med Sci. 2019; 26: 38-52, CrossRef.

Hilbertina N, Abdullah M, Siregar NC, Wanandi SI. Determination of cancer‑associated fibroblast and stromal phenotypes as novel prognostic factors for colorectal carcinomas associated with tumor budding. J Nat Sci Biol Med. 2019; 10: S68-72, article.

Herrera M, Islam ABMMK, Herrera A, Martín P, García V, Silva J, et al. Functional heterogeneity of cancer-associated fibroblasts from human colon tumors shows specific prognostic gene expression signature. Clin Cancer Res. 2013; 19: 5914-26, CrossRef.

Tao L, Huang G, Song H, Chen Y, Chen L. Cancer associated fibroblasts: An essential role in the tumor microenvironment. Oncol Lett. 2017; 14: 2611-20, CrossRef.

Fiori ME, Di Franco S, Villanova L, Bianca P, Stassi G, De Maria R. Cancer-associated fibroblasts as abettors of tumor progression at the crossroads of EMT and therapy resistance. Mol Cancer. 2019; 18: 1-16, CrossRef.

Pradella D, Naro C, Sette C, Ghigna C. EMT and stemness: flexible processes tuned by alternative splicing in development and cancer progression. Mol Cancer. 2017; 16: 1-19, CrossRef.

Yu Y, Xiao CH, Tan LD, Wang QS, Li XQ, Feng YM. Cancer-associated fibroblasts induce epithelial–mesenchymal transition of breast cancer cells through paracrine TGF-β signalling. Br J Cancer. 2014; 110: 724-32, CrossRef.

Donnarumma E, Fiore D, Nappa M, Roscigno G, Adamo A, Iaboni M, et al. Cancer-associated fibroblasts release exosomal microRNAs that dictate an aggressive phenotype in breast cancer. 2017; 8: 19592-608, CrossRef.

da Cunha BR, Domingos C, Stefanini ACB, Henrique T, Polachini GM, Castelo-Branco P, et al. Cellular interactions in the tumor microenvironment: the role of secretome. J Cancer. 2019; 10: 4574-87, CrossRef.

Purnamawati, Pawitan JA, Rachman A, Liem IK, Wanandi SI. Secretomes of adipose and umbilical cord-derived stem cells affect ALDH1A1 expression in breast cancer stem cells. Adv Sci Lett. 2017; 23: 6701-4, CrossRef.

Wanandi SI, Wideani G, Zainuri M, Putri RR, Bachtiar EW, Jusman SWA. Evaluation of stemness and proliferation of human breast cancer stem cells (ALDH+) supplemented with heat-activated TGF-beta1 in the secretomes of stem cells from human exfoliated deciduous teeth (SHED). Ann Oncol. 2018; 29(Suppl 8): viii1–viii13, CrossRef.

Zhuang J, Lu Q, Shen B, Huang X, Shen L, Zheng X, et al. TGFβ1 secreted by cancer-associated fibroblasts induces epithelial-mesenchymal transition of bladder cancer cells through lncRNA-ZEB2NAT. Sci Rep. 2015; 5: 11924, CrossRef.

Sun Y, Fan X, Zhang Q, Shi X, Xu G, Zou C. Cancer-associated fibroblasts secrete FGF-1 to promote ovarian proliferation, migration, and invasion through the activation of FGF-1/FGFR4 signaling. Tumor Biol. 2017; 39: 101042831771259, CrossRef.

Peng Q, Zhao L, Hou Y, Sun Y, Wang L, Luo H, et al. Biological characteristics and genetic heterogeneity between carcinoma-associated fibroblasts and their paired normal fibroblasts in human breast cancer. PLoS One. 2013; 8: e60321, CrossRef.




DOI: https://doi.org/10.18585/inabj.v12i4.1295

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