Open AccessUpregulation of MiR-340-5p Reverses Cisplatin Sensitivity by Inhibiting the Expression of CDK6 in HepG2 Cells
Author(s) -
Sha Tian,
Zhuo Liu,
Qing Zhou,
Ruoxi Wu,
Xiaodi Huang,
Zicheng Liang,
Zhen Zhang,
Xuefei Tian
Publication year - 2021
Publication title -
folia biologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.165
H-Index - 23
eISSN - 1734-9168
pISSN - 0015-5497
DOI - 10.3409/fb_69-2.08
Subject(s) - transfection , cisplatin , cyclin dependent kinase 6 , cell cycle , clonogenic assay , apoptosis , cell growth , cancer research , flow cytometry , chemistry , gene silencing , cell cycle checkpoint , cancer cell , microbiology and biotechnology , cell culture , biology , cancer , chemotherapy , cyclin dependent kinase 2 , biochemistry , genetics , gene
Cisplatin (CDDP) has been successfully used in chemotherapy for liver cancer. However, the development of CDDP resistance in HepG2 cells usually leads to relapse and a worsening prognosis. MiR-340-5p has attracted much attention because of its ability to affect cell resistance. This project is intended to explore the role of miR-340-5p and CDK6 in CDDP-R HepG2 cells and provide new ideas for the treatment of liver cancer. A dual-luciferase reporter assay was used to confirm the targeting relationship between miR-340-5p and CDK6. We constructed a CDDP-resistant model of HepG2 cells to examine the effect of miR-340-5p on the drug sensitivity of HepG2 cells. CDDP-R HepG2 cells were transfected with miR-340-5p overexpression plasmid and CDK6 silencing plasmid. QRT-PCR was used to detect the expression of miR-340-5p and CDK6. A western blot was performed to determine the expression of CDK6, CyclinD1, and CyclinD2. CCK-8, flow cytometry, TUNEL and Clonogenic assays were also carried out to detect CDDP-R HepG2 cells. There is a targeting relationship between miR-340-5p and CDK6. The drug resistance of CDDP-R HepG2 cells was significantly higher than that of CDDP-S HepG2 cells. CDDP-R HepG2 cells transfected with both miR-340-5p overexpressing plasmid and CDK6 silencing plasmid showed a lower proliferation ability, cell cycle arrest in the G0/G1 phase, and lower drug resistance compared with single CDDP-R HepG2 cells. Overexpression of miR-340-5p aggravated CDDP-R HepG2 cells' apoptosis and inhibited cell viability. Overexpression of miR-340-5p could reverse the sensitivity of HepG2 cells to CDDP by inhibiting the expression of CDK6 in HepG2 cells.