Knock-down of long non-coding RNA GK-IT1 inhibits proliferation and promotes apoptosis in esophageal carcinoma cells

Di-san junyi daxue xuebao

CHEN Li¹  YANG Xin¹ 

[1] Department of Cardiothoracic Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China;

关键词: long non-coding rna;    gk-it1;    esophageal squamous cell carcinoma;    cell proliferation;    apoptosis;   

DOI  :  10.16016/j.1000-5404.202008083

来源: DOAJ

【 摘 要 】

Objective To investigate the expression of long non-coding (lnc) RNA GK-IT1 in esophageal squamous cell carcinoma (ESCC) and explore its effect on the proliferation and apoptosis of ESCC cell lines. Methods Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was carried out to detect the expression of lncRNA GK-IT1 in 27 pairs of ESCC tissues and adjacent normal tissues from the ESCC patients who had not received chemotherapy yet, and in different ESCC cell lines (KYSE-150, KYSE-510, Eca-109, TE-1 and TE-10) and normal human esophageal epithelioid cell line Het-1a, respectively. After small interference RNA (siRNA) of GK-IT1 was designed and constructed, and then transfected into TE-1 and TE-10 cells respectively to knockdown GK-IT1. Plate colony formation assay, EDU analysis and cell counting kit (CCK-8) assay were used to determine the proliferation of the cells. Flow cytometry was employed to detect the variations of apoptosis and cell cycle. Fluorescence in situ hybridization (FISH) was used to explore the subcellular localization of GK-IT1, and Western blotting was performed to verify the expression of cell cycle-related proteins. Results The expression of GK-IT1 was significantly higher in the ESCC tissues when compared to para-cancerous tissues (P < 0.000 1). GK-IT1 knockdown cells were successfully constructed in TE-1 and TE-10 cells, and knock-down of GK-IT1 could significantly inhibit the proliferation (P < 0.01) and increase the apoptotic rate of the cells (P < 0.01). The results of FISH assay indicated that GK-IT1 was mainly distributed in the cytoplasm. Flow cytometry showed that GK-IT1 knockdown induced the cells arrested in G1/S phase. Finally, bioinformatics analysis suggested that GK-IT1 may bind to CDK4/CCND1 protein, and Western blotting results revealed that the expression of CDK4 and CCND1 was decreased after GK-IT1 knockdown (P < 0.05). Conclusion Knockdown of GK-IT1 can induce cell cycle arrest, inhibit proliferation and promote apoptosis in ESCC cells. It may be used as a potential therapeutic target in treatment for ESCC.

【 授权许可】

Unknown