Open Access
ARTICLE
NET1 Enhances Proliferation and Chemoresistance in Acute Lymphoblastic Leukemia Cells
Hongbo Sun*1,
Zhifu Zhang*1,
Wei Luo*, Junmin Liu*, Ye Lou†, Shengmei Xia‡
* Department of Hematology, Shenzhen Longhua People’s Hospital, Shenzhen, P.R. China
† Department of Hematology, Daqing Oilfield General Hospital, Daqing, P.R. China
‡ Department of Neurology, Shenzhen Longhua People’s Hospital, Shenzhen, P.R. China
Oncology Research 2019, 27(8), 935-944. https://doi.org/10.3727/096504019X15555388198071
Abstract
Acute lymphoblastic leukemia (ALL) is the most prevalent of pediatric cancers. Neuroepithelial cell-transforming 1
(NET1) has been associated with malignancy in a number of cancers, but the role of NET1 in ALL development is unclear. In the present study, we investigated the effect of NET1 gene in ALL cell proliferation and
chemoresistance. We analyzed GEO microarray data comparing bone marrow expression profiles of pediatric
B-cell ALL samples and those of age-matched controls. MTT and colony formation assays were performed
to analyze cell proliferation. ELISA assays, Western blot analyses, and TUNEL staining were used to detect
chemoresistance. We confirmed that NET1 was targeted by miR-206 using Western blot and luciferase reporter
assays. We identified NET1 gene as one of the most significantly elevated genes in pediatric B-ALL. MTT
and colony formation assays demonstrated that NET1 overexpression increases B-ALL cell proliferation in
Nalm-6 cells. ELISA assays, Western blot analyses, and TUNEL staining showed that NET1 contributes to
ALL cell doxorubicin resistance, whereas NET1 inhibition reduces resistance. Using the TargetScan database,
we found that several microRNAs (miRNAs) were predicted to target NET1, including microRNA-206 (miR-
206), which has been shown to regulate cancer development. To determine whether miR-206 targets NET1
in vitro, we transfected Nalm-6 cells with miR-206 or its inhibitor miR-206-in. Western blot assays showed
that miR-206 inhibits NET1 expression and miR-206-in increases NET1 expression. Luciferase assays using
wild-type or mutant 3 -untranslated region (3 -UTR) of NET1 confirmed these findings. We ultimately found
that miR-206 inhibits B-ALL cell proliferation and chemoresistance induced by NET1. Taken together, our
results provide the first evidence that NET1 enhances proliferation and chemoresistance in B-ALL cells and
that miR-206 regulates these effects by targeting NET1. This study therefore not only contributes to a greater
understanding of the molecular mechanisms underlying B-ALL progression but also opens the possibility for
developing curative interventions.
Keywords
Cite This Article
Sun, H., Zhang,
. Z., Luo,
. W., Liu, J., Lou, Y. et al. (2019). NET1 Enhances Proliferation and Chemoresistance in Acute Lymphoblastic Leukemia Cells.
Oncology Research, 27(8), 935–944. https://doi.org/10.3727/096504019X15555388198071