Spheroid-Derived Cells From Renal Adenocarcinoma Have Low Telomerase Activity and High Stem-Like and Invasive Characteristics

Leili Saeednejad Zanjani; Zahra Madjd; Arezoo Rasti; Mojgan Asgari; Maryam Abolhasani; Kevin J Tam; Raheleh Roudi; Gunhild Mari Mælandsmo; Øystein Fodstad; Yvonne Andersson

doi:10.3389/fonc.2019.01302

Spheroid-Derived Cells From Renal Adenocarcinoma Have Low Telomerase Activity and High Stem-Like and Invasive Characteristics

Front Oncol. 2019 Dec 4:9:1302. doi: 10.3389/fonc.2019.01302. eCollection 2019.

Authors

Leili Saeednejad Zanjani^{1

2}, Zahra Madjd^{1

3

4}, Arezoo Rasti^{1

5}, Mojgan Asgari^{1

6}, Maryam Abolhasani^{1

6}, Kevin J Tam⁴, Raheleh Roudi¹, Gunhild Mari Mælandsmo⁷, Øystein Fodstad⁷, Yvonne Andersson⁷

Affiliations

¹ Oncopathology Research Center, Iran University of Medical Sciences (IUMS), Tehran, Iran.
² Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran.
³ Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
⁴ Department of Urologic Sciences, Vancouver Prostate Center, University of British Columbia, Vancouver, BC, Canada.
⁵ Department of Basic Sciences/Medical Surgical Nursing, Faculty of Nursing and Midwifery, Tehran University of Medical Sciences, Tehran, Iran.
⁶ Hasheminejad Kidney Center, Iran University of Medical Sciences (IUMS), Tehran, Iran.
⁷ Department of Tumor Biology, Oslo University Hospital, The Norwegian Radium Hospital, Oslo, Norway.

Abstract

Cancer stem cells (CSCs) are a theorized small subpopulation of cells within tumors thought to be responsible for metastasis, tumor development, disease progression, treatment-resistance, and recurrence. The identification, isolation, and biological characterization of CSCs may therefore facilitate the development of efficient therapeutic strategies targeting CSCs. This study aims to compare the biology and telomerase activity of CSCs to parental cells (PCs) in renal cancer. Renal CSCs were enriched from the ACHN cell line using a sphere culture system. Spheroid-derived cells (SDCs) and their adherent counterparts were compared with respect to their colony and sphere formation, expression of putative CSC markers, tumorigenicity in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice, and invasiveness. The expression of genes associated with CSCs, stemness, EMT, apoptosis, and ABC transporters was also compared between the two populations using quantitative real-time PCR (qRT-PCR). Finally, telomerase activity, hTERT expression, and sensitivity to MST-312, a telomerase inhibitor, was investigated between the two populations. We demonstrated that a subpopulation of ACHN cells was capable of growing as spheroids with many properties similar to CSCs, including higher clonogenicity, superior colony- and sphere-forming ability, and stronger tumorigenicity and invasiveness. In addition, SDCs demonstrated a higher expression of markers for CSCs, stemness, EMT, apoptosis, and ABC transporter genes compared to PCs. The expression of hTERT and telomerase activity in SDCs was significantly lower than PCs; however, the SDC population was more sensitive to MST-312 compared to PCs. These findings indicate that the SDC population exhibits stem-like potential and invasive characteristics. Moreover, the reduced expression of hTERT and telomerase activity in SDCs demonstrated that the expressions of hTERT and telomerase activity are not always higher in CSCs. Our results also showed that MST-312 treatment inhibited SDCs more strongly than PCs and may therefore be useful as a complementary targeted therapy against renal CSCs in the future.

Keywords: MST-312; cancer stem cells; hTERT; renal cell carcinoma; telomerase activity.