Long noncoding RNA LINC00968 inhibits proliferation, migration and invasion of lung adenocarcinoma through targeting miR-22-5p/CDC14A axis.

Yang Wu,Tianli Pei,Chao Wu,Liyuan Zhang,Yuanyuan Hu,Xuzhao Bian,Xinpeng Han

doi:10.1007/s13205-021-02981-8

Abstract

Lung adenocarcinoma (LUAD) is a high aggressive human cancer which usually diagnosed at advanced stages. Accumulating evidences indicate that long noncoding RNAs (lncRNAs) are crucial participants in LUAD progression. In the present study, we found that lncRNA LINC00968 was significantly down-regulated in LUAD tissues and cell lines. LINC00968 level was positively correlated to survival rate, and negatively correlated to tumor node metastasis (TNM) stage, tumor size and lymph node metastasis of LUAD patients. We over-expressed LINC00968 in LUAD cells using lentivirus, inhibited proliferation and cell cycle arrest at G1 phase were detected. LINC00968 over-expression also suppressed migration, invasion and epithelial mesenchymal transition. We further validated that LINC00968 localized in cytoplasm and acted as an upstream regulator of microRNA miR-22-5p, which was up-regulated in LUAD tissues and cell lines. Besides, elevated miR-22-5p expression abolished the effect of LINC00968 over-expression on LUAD progression including in vivo tumor growth. In addition, we first validated that cell division cycle 14A (CDC14A), which was down-regulated in LUAD tissues, was a downstream target of miR-22-5p. We over-expressed CDC14A in LUAD cells and miR-22-5p induced LUAD progression was partially reversed. In conclusion, our study demonstrated that LINC00968 inhibited proliferation, migration and invasion of LUAD by sponging miR-22-5p and further restoring CDC14A. This novel regulatory axis might provide us with promising diagnostic and therapeutic target in LUAD treatment.

Highlights

Lung adenocarcinoma (LUAD) is a high aggressive human cancer which usually diagnosed at advanced stages
We found that long noncoding RNAs (lncRNAs) LINC00968 was significantly down-regulated in LUAD tissues and cell lines
LINC00968 level was positively correlated to survival rate, and negatively correlated to tumor node metastasis stage, tumor size and lymph node metastasis of LUAD patients

Summary

Methods

The mRNA levels of LINC00968, miR-22-5p and cell division cycle 14A (CDC14A) were measured using quantitative real-time PCR. All LUAD tissues and adjacent non-tumor lung tissues (n = 60) were collected with the consent of patients. All experiment regarding clinical samples were performed in accordance with the ethical guidelines of the Declaration of Helsinki and approved by the Ethics Committee of International Medical Center Hospital. Normal lung epithelial cell line BEAS2B, LUAD cell lines A549 and H1975 were purchased from Procell (China). LUAD cell lines HCC2935 and PC-9 were obtained from the American type culture collection (ATCC) cell bank. BEAS2B, A549 and H1975 cells were cultured with dulbecco's modified eagle medium (DMEM) containing 10% fatal bovine serum (FBS). HCC2935 and PC-9 cells were maintained in Roswell Park Memorial Institute-1640 (RPMI-1640) medium with 10% FBS. LINC00968 was inserted into lentiviral vector pRLenti-CMV-MCS-3FLAG (OBIO, China) for over-expression. Cells were transfected with mimics, agomirs and pcDNA3.1 vectors using lipofectamine2000 according to the manufactures’ instruction

Results

Discussion

Conclusion