Effects of leukemia inhibitory factor receptor overexpression on stemness maintenance and lung metastasis in vivo of thyroid cancer stem cells

doi:10.3969/j.issn.2095-4344.0467

Abstract

Abstract:

BACKGROUND: Thyroid cancer stem cells are essential to the recurrence and metastasis of thyroid carcinoma. Leukemia inhibitory factor receptor (LIFR) shows a downward trend in a variety of malignant tumors, and its overexpression can inhibit the recurrence and metastasis of malignant tumors.
OBJECTIVE: To explore the effect of LIFR on the stemness maintenance and lung metastasis of thyroid cancer stem cells in vivo.
METHODS: Primary thyroid cancer cells TCLM were isolated from the lung metastases of a metastatic thyroid cancer patient. Serum-free suspension culture was used to form tumor cell balls. Flow cytometry was used to screen CD133⁺ phenotype of metastatic thyroid cancer stem cell subpopulation TCLM-S. The overexpressed recombinant lentiviral plasmid containing LIFR and its negative control containing the empty plasmid were infected into thyroid cancer stem cells TCLM-S at the ratio of virus/cell number=20, and screened with 2.0 mg/L puromycin to construct TCLM-S_LIFR and TCLM-S_control stem cells which stably expressed LIFR and its control. Real-time quantitative PCR (qRT-PCR) was used to detect the expression of LIFR in TCLM-S_LIFR and TCLM-S_controlstem cells. Flow cytometry was used to detect the percentage of CD133⁺ phenotype cell subsets, western blot assay was used to detect the expression of tumor stemness related factors SOX2, Oct4, Nanog and tumor invasion and metastasis related proteins E-cadherin, matrix metalloproteinase (MMP)-2, MMP-7 in TCLM-S_LIFR and TCLM-Scontrol stem cells. TCLM-S_LIFR and TCLM-S_control stem cells were respectively injected into BALB/c nude mice by tail vein, and the lung metastasis model of thyroid cancer stem cells was constructed. The effect of LIFR overexpression on lung metastasis was observed.
RESULTS AND CONCLUSION: Compared with TCLM-S_controlcells, the expression of LIFR in TCLM-S_LIFR cells was significantly increased, the proportion of CD133⁺ phenotype stem cell subsets was significantly decreased, the expression of SOX2, Oct4 and Nanog were significantly decreased, the expression of E-cadherin was significantly increased, and the expression of MMP-2 and MMP-7 was significantly decreased. Moreover, the number of lung metastasis in nude mice given TCLM-S_LIFR cells was significantly decreased as compared with those given TCLM-Scontrol cells. To conclude, LIFR overexpression can decrease the stemness and ability of lung metastasis in vivo.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Neoplastic Stem Cells, Thyroid Neoplasms, Leukemia Inhibitory Factor, Neoplasm Metastasis, Tissue Engineering

CLC Number:

R394.2

Zhang Zhen-hua, Su Zi-jie, Kan Yun-zhen, Liu Qiu-yu. Effects of leukemia inhibitory factor receptor overexpression on stemness maintenance and lung metastasis in vivo of thyroid cancer stem cells[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(9): 1376-1381.

Figures/Tables 1

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