The study of siRNA interference after laryngeal cancer Hep-2 cells to cisplatin sensitivity of β-catenin gene expression.
- Author:
Feng YU
;
Xin HUANG
;
Maomao AI
;
Ying LIN
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cell Line, Tumor;
Cisplatin;
pharmacology;
Drug Resistance, Neoplasm;
Humans;
Laryngeal Neoplasms;
genetics;
RNA Interference;
RNA, Messenger;
RNA, Small Interfering;
beta Catenin;
genetics
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2015;29(13):1143-1147
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the changes of laryngeal cancer Hep-2 cells to cisplatin chemosensitivity after the interference of siRNA of β-catenin gene expression.
METHOD:Using a small interference RNA (siRNA) technology interfere β-catenin gene of Hep-2 cells . The mRNA and protein levels of β-catenin in the Hep-2 cells of different groups were detected by qPCR and Western blot. It was divided into siRNA-β-catenin-Hep-2 siRNA group, β-catenin-Neg negative control group and blank control group. Cell proliferation inhibition rate of different concentrations of cisplatin on three groups was detected by MTT assay. Calculate the 50% inhibitory effective concentration IC50 value. Check the change of three groups of cells' apoptosis rate by flow cytometry after the same concentrations of cisplatin stimulation.
RESULT:β-catenin-siRNA interference fragment can specifically reduce the expression levels of β-catenin mRNA and protein. qPCR illustrated the expression of mRNA in β-catenin-siR-NA-Hep-2 interference group decreased 70% (P < 0.05) compared with the control group, Western blot results showed that the β-catenin protein expression of interference group (0. 545 ± 0.111) decreased significantly compared with blank control group (1.507 ± 0.139) and negative control group (1.429 ± 0.089), P < 0.05. The IC50 calculation software showed that IC50 of cisplatin on β-catenin-siRNA IC50 interference group is (5.81 ± 0.46)μg/ml, the blank control group is (10.10 ± 1.01) μg/ml, the difference between the two groups has statistical signifi- cance (P < 0.01). Cell apoptosis rate of β-catenin-siRNA interference group was (26.15 ± 0.60)%, significantly higher than the control group (14.16 ± 0.05)%, P < 0.05.
CONCLUSION:To interfere the expression of β-catenin can effectively enhance the sensitivity of laryngeal cancer cells to chemotherapeutic drugs cisplatin. It provides a theoretical support for the reduction of laryngeal cancer chemotherapy drug cisplatin dosage.
