The DNMT1/miR-34a Axis Is Involved in the Stemness of Human Osteosarcoma Cells and Derived Stem-Like Cells
The DNA methyltransferase 1 (DNMT1)/miR-34a axis promoted carcinogenesis of various types of cancers. However, no literature reported its contribution to the stemness of osteosarcoma cancer stem-like cells (OSLCs). We sought to determine whether the DNMT1/miR-34a axis facilitates the stemness of OSL...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2019-01-01
|
| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2019/7028901 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832545576278818816 |
|---|---|
| author | Xiao Liang Chang Xu Wanchun Wang Xiang Li |
| author_facet | Xiao Liang Chang Xu Wanchun Wang Xiang Li |
| author_sort | Xiao Liang |
| collection | DOAJ |
| description | The DNA methyltransferase 1 (DNMT1)/miR-34a axis promoted carcinogenesis of various types of cancers. However, no literature reported its contribution to the stemness of osteosarcoma cancer stem-like cells (OSLCs). We sought to determine whether the DNMT1/miR-34a axis facilitates the stemness of OSLCs. We here revealed the higher DNMT1 activity and expression, lower miR-34a expression with high methylation of its promoter, and stronger stemness of OSLCs, as manifested by elevated sphere and colony formation capacities; CD133, CD44, ABCG2, Bmi1, Sox2, and Oct4 protein amounts in vitro; and carcinogenicity in a nude mouse xenograft model, when compared to the parental U2OS cells. 5-Azacytidine (Aza-dC) repressed DNMT1 activation and upregulated miR-34a expression by promoter demethylation and suppressed the stemness of OSLCs in a dose-dependent manner. DNMT1 knockdown increased miR-34a and reduced the stemness of OSLCs. Transfection with a miR-34a mimic repressed the stemness of OSLCs but did not alter DNMT1 activity and expression. Conversely, DNMT1 overexpression declined miR-34a levels, promoting the stemness of U2OS cells. Transfection with a miR-34a inhibitor enhanced the stemness of U2OS cells, without affecting the DNMT1 activity and expression. Importantly, reexpression of miR-34a could rescue the effects of DNMT1 overexpression on miR-34a inhibition as well as the stemness promotion without affecting the activity and expression of DNMT1. Our results revealed that aberrant activation of DNMT1 caused promoter methylation of miR-34a, leading to miR-34a underexpression, and the role of the DNMT1/miR-34a axis in promoting and sustaining the stemness of OSLCs. |
| format | Article |
| id | doaj-art-57de8c3eac3c46218bb2952b3457580d |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-57de8c3eac3c46218bb2952b3457580d2025-02-03T07:25:17ZengWileyStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/70289017028901The DNMT1/miR-34a Axis Is Involved in the Stemness of Human Osteosarcoma Cells and Derived Stem-Like CellsXiao Liang0Chang Xu1Wanchun Wang2Xiang Li3Department of Orthopaedics, The Second Xiangya Hospital, Central South University, 139 Renmin Road, Changsha, Hunan 410011, ChinaDepartment of Preclinical Medicine, Medical College, Hunan Normal University, Changsha, Hunan 410013, ChinaDepartment of Orthopaedics, The Second Xiangya Hospital, Central South University, 139 Renmin Road, Changsha, Hunan 410011, ChinaDepartment of Preclinical Medicine, Medical College, Hunan Normal University, Changsha, Hunan 410013, ChinaThe DNA methyltransferase 1 (DNMT1)/miR-34a axis promoted carcinogenesis of various types of cancers. However, no literature reported its contribution to the stemness of osteosarcoma cancer stem-like cells (OSLCs). We sought to determine whether the DNMT1/miR-34a axis facilitates the stemness of OSLCs. We here revealed the higher DNMT1 activity and expression, lower miR-34a expression with high methylation of its promoter, and stronger stemness of OSLCs, as manifested by elevated sphere and colony formation capacities; CD133, CD44, ABCG2, Bmi1, Sox2, and Oct4 protein amounts in vitro; and carcinogenicity in a nude mouse xenograft model, when compared to the parental U2OS cells. 5-Azacytidine (Aza-dC) repressed DNMT1 activation and upregulated miR-34a expression by promoter demethylation and suppressed the stemness of OSLCs in a dose-dependent manner. DNMT1 knockdown increased miR-34a and reduced the stemness of OSLCs. Transfection with a miR-34a mimic repressed the stemness of OSLCs but did not alter DNMT1 activity and expression. Conversely, DNMT1 overexpression declined miR-34a levels, promoting the stemness of U2OS cells. Transfection with a miR-34a inhibitor enhanced the stemness of U2OS cells, without affecting the DNMT1 activity and expression. Importantly, reexpression of miR-34a could rescue the effects of DNMT1 overexpression on miR-34a inhibition as well as the stemness promotion without affecting the activity and expression of DNMT1. Our results revealed that aberrant activation of DNMT1 caused promoter methylation of miR-34a, leading to miR-34a underexpression, and the role of the DNMT1/miR-34a axis in promoting and sustaining the stemness of OSLCs.http://dx.doi.org/10.1155/2019/7028901 |
| spellingShingle | Xiao Liang Chang Xu Wanchun Wang Xiang Li The DNMT1/miR-34a Axis Is Involved in the Stemness of Human Osteosarcoma Cells and Derived Stem-Like Cells Stem Cells International |
| title | The DNMT1/miR-34a Axis Is Involved in the Stemness of Human Osteosarcoma Cells and Derived Stem-Like Cells |
| title_full | The DNMT1/miR-34a Axis Is Involved in the Stemness of Human Osteosarcoma Cells and Derived Stem-Like Cells |
| title_fullStr | The DNMT1/miR-34a Axis Is Involved in the Stemness of Human Osteosarcoma Cells and Derived Stem-Like Cells |
| title_full_unstemmed | The DNMT1/miR-34a Axis Is Involved in the Stemness of Human Osteosarcoma Cells and Derived Stem-Like Cells |
| title_short | The DNMT1/miR-34a Axis Is Involved in the Stemness of Human Osteosarcoma Cells and Derived Stem-Like Cells |
| title_sort | dnmt1 mir 34a axis is involved in the stemness of human osteosarcoma cells and derived stem like cells |
| url | http://dx.doi.org/10.1155/2019/7028901 |
| work_keys_str_mv | AT xiaoliang thednmt1mir34aaxisisinvolvedinthestemnessofhumanosteosarcomacellsandderivedstemlikecells AT changxu thednmt1mir34aaxisisinvolvedinthestemnessofhumanosteosarcomacellsandderivedstemlikecells AT wanchunwang thednmt1mir34aaxisisinvolvedinthestemnessofhumanosteosarcomacellsandderivedstemlikecells AT xiangli thednmt1mir34aaxisisinvolvedinthestemnessofhumanosteosarcomacellsandderivedstemlikecells AT xiaoliang dnmt1mir34aaxisisinvolvedinthestemnessofhumanosteosarcomacellsandderivedstemlikecells AT changxu dnmt1mir34aaxisisinvolvedinthestemnessofhumanosteosarcomacellsandderivedstemlikecells AT wanchunwang dnmt1mir34aaxisisinvolvedinthestemnessofhumanosteosarcomacellsandderivedstemlikecells AT xiangli dnmt1mir34aaxisisinvolvedinthestemnessofhumanosteosarcomacellsandderivedstemlikecells |