Suppression of CLEC3A inhibits osteosarcoma cell proliferation and promotes their chemosensitivity through the AKT1/mTOR/HIF1α signaling pathway

Ren,Chong; Pan,Runsang; Hou,Lisong; Wu,Huaping; Sun,Junkang; Zhang,Wenguang; Tian,Xiaobin; Chen,Houping

doi:10.3892/mmr.2020.10986

Suppression of CLEC3A inhibits osteosarcoma cell proliferation and promotes their chemosensitivity through the AKT1/mTOR/HIF1α signaling pathway

Authors:
- Chong Ren
- Runsang Pan
- Lisong Hou
- Huaping Wu
- Junkang Sun
- Wenguang Zhang
- Xiaobin Tian
- Houping Chen
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Affiliations: Department of Orthopedics, Guiyang Maternal and Child Health-Care Hospital, Guiyang, Guizhou 550000, P.R. China, Department of Orthopedics, Clinical Medical College of Guizhou Medical University, Guiyang, Guizhou 550000, P.R. China
Published online on: February 13, 2020 https://doi.org/10.3892/mmr.2020.10986
Pages: 1739-1748
Copyright: © Ren et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Osteosarcoma (OS) is a primary malignant tumor that occurs in bone, and mainly affects children and adolescents. C‑type lectin domain family 3 member A (CLEC3A) is a member of the C‑type lectin superfamily, which regulates various biological functions of cells. The present study aimed to identify the effects and related mechanisms of CLEC3A in the proliferation and chemosensitivity of OS cells. The expression of CLEC3A in OS was analyzed using the Gene Expression Omnibus data profile GSE99671, and its expression in OS samples was verified using reverse transcription‑quantitative PCR (RT‑qPCR) and immunohistochemical staining. The relationship between the expression of CLEC3A and clinical traits in patients with OS was also analyzed, including age, tumor size, TNM stage and lymph node metastasis. Cell Counting Kit‑8 assays, colony formation assays and cell cycle distribution analysis were used to determine the roles of CLEC3A in the proliferation and chemosensitivity of OS cells. Finally, RT‑qPCR and western blotting were used to demonstrate the relationship between CLEC3A and the AKT1/mTOR/hypoxia‑inducible factor 1‑α (HIF1α) pathway. Both the mRNA and protein expression levels of CLEC3A were increased in OS tissues compared with adjacent non‑tumor tissues, and this was positively associated with TNM stage and lymph node metastasis. The genetic knockdown of CLEC3A with small interfering RNA decreased OS cell proliferation and colony formation, and induced G1 phase arrest, whereas the overexpression of CLEC3A increased OS cell proliferation and colony formation, and alleviated G1 phase arrest. The suppression of CLEC3A also promoted enhanced the chemosensitivity of OS cells to doxorubicin (DOX) and cisplatin (CDDP); it also inhibited the expression of AKT1, mTOR and HIF1α, further to the nuclear localization of HIF1α, and HIF1α target gene expression levels, including VEGF, GLUT1 and MCL1 were also decreased. Furthermore, treatment with the AKT activator SC79 blocked the inhibitory effects of CLEC3A silencing in OS cells. In conclusion, these findings suggested that CLEC3A may function as an oncogene in OS, and that the suppression of CLEC3A may inhibit OS cell proliferation and promote chemosensitivity through the AKT1/mTOR/HIF1α signaling pathway.

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