Open AccessLong non‐coding RNA POU6F2‐AS2 promotes cell proliferation and drug resistance in colon cancer by regulating miR‐377/BRD4
Author(s) -
Xu Guangru,
Zhu Hongxing,
Xu Jinhua,
Wang Yan,
Zhang Yang,
Zhang Minghui,
Zhu Dichao
Publication year - 2020
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/jcmm.15070
Subject(s) - cell growth , cell cycle , flow cytometry , microbiology and biotechnology , biology , apoptosis , transfection , colorectal cancer , cancer research , cell , suppression subtractive hybridization , cell culture , chemistry , cancer , messenger rna , gene , cdna library , biochemistry , genetics
The aim of this study was to explore the molecular mechanism of lncRNA POU6F2‐AS2 in proliferation and drug resistance of colon cancer. Total paired 70 colon cancer and adjacent normal tissues were collected from colon cancer patients. Colon cancer and normal colonic epithelial cells were purchased. POU6F2‐AS2 was up‐ or down‐expressed by vectors. LC50 of all cell lines before and after transfection with these plasmids was detected. qRT‐PCR was used to detect the expression of POU6F2‐AS2, miR‐377 and BRD4 before or after transfection. In situ hybridization was also undertaken to detect the level of POU6F2‐AS2. Different concentrations of 5‐Fu (0, 1, 2.5, 5, 10, 20, 40 and 80 μg/mL) were used for 5‐FU insensitivity assay. CCK‐8 and crystal violet staining assay were used for detecting cell proliferation, and flow cytometry was used for identifying cell cycle distribution and apoptosis. In order to detect the fragmented DNA in apoptotic cells, TUNEL assay was used. RNA pull‐down assay and luciferase reporter assay were used to verify the binding site. Rescue assay confirmed the subtractive effect of miR‐377 inhibitors. POU6F2‐AS2 was highly expressed in colon cancer, which was associated with clinical pathology. Up‐regulated POU6F2‐AS2 promoted cell proliferation and cell cycle of colon cancer cells. Overexpression of POU6F2‐AS2 inhibited the expression of miR‐377 and then up‐regulated the expression of BRD4. Up‐regulated BRD4 ultimately promoted cell proliferation and cell survival Down‐regulated POU6F2‐AS2 showed enhanced sensitivity of 5‐FU. POU6F2‐AS2 promoted cell proliferation and drug resistance in colon cancer by regulating miR‐377/BRD4 gene.