Apolipoprotein M could inhibit growth and metastasis of SMMC7721 cells via vitamin D receptor signaling
Received 24 January 2019
Accepted for publication 5 April 2019
Published 30 April 2019 Volume 2019:11 Pages 3691—3701
Checked for plagiarism Yes
Review by Single-blind
Peer reviewers approved by Dr Melinda Thomas
Peer reviewer comments 2
Editor who approved publication: Dr Kenan Onel
Miaomei Yu,1 Lili Pan,1 Chen Sang,2 Qinfeng Mu,1 Lu Zheng,1 Guanghua Luo,1 Ning Xu3
1Comprehensive Laboratory, the Third Affiliated Hospital of Soochow University, Changzhou 213003, People’s Republic of China; 2Department of Cardiothoracic Surgery, the Third Affiliated Hospital of Soochow University, Changzhou 213003, People’s Republic of China; 3Section of Clinical Chemistry and Pharmacology, Institute of Laboratory Medicine, Lunds University, Lund S‑22185, Sweden
Objective: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors with high mortality-to-incidence ratios. Apolipoprotein M (ApoM), a member of the apolipoprotein family, is mainly synthesized in the liver, whereas its role in HCC has not been elucidated. Here, we examined the effect of ApoM on the biological behavior of HCC cells and the possible mechanisms.
Methods: We used CRISPR/Cas9 technology to knock out ApoM in SMMC7721 cells. Differentially expressed genes before and after ApoM knockout (KO) were analyzed by GeneChip microarrays and confirmed by qRT-PCR. Cell assays of proliferation, apoptosis, migration and invasion were performed in SMMC7721 cells, and the expression of epithelial–mesenchymal transition (EMT) markers was performed by western blot. And we performed functional recovery experiments by overexpressing vitamin D receptor (VDR) in SMMC7721.
Results: The ApoM-KO SMMC7721 cell line was successfully constructed using the CRISPR/Cas9 technology. Our results showed that silencing ApoM suppressed apoptosis and promoted proliferation, migration, invasion and EMT of SMMC7721 cells. The microarray data revealed that a total of 1,868 differentially expressed genes were identified, including VDR. The qRT-PCR and western blot verification results demonstrated that knocking out ApoM could significantly reduce the expression of VDR. The functional recovery experiments indicated that VDR overexpression could offset the inhibition of cell apoptosis and the promotion of cell proliferation, migration, invasion and EMT caused by knocking out ApoM in SMMC7721 cells.
Conclusion: ApoM could function as a tumor suppressor to inhibit the growth and metastasis of SMMC7721 cells via VDR signaling in HCC.
Keywords: apolipoprotein M, CRISPR/Cas9, hepatocellular carcinoma cells, GeneChip microarrays, vitamin D receptor
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