The effect of TFF3 on the proliferation and migration of papillary thyroid carcinoma K1 cell.
- Author:
Xiaochun ZHENG
;
Tingting ZHANG
;
Jingfang WU
;
Wenjing ZHANG
;
Jing ZHANG
;
Baozhi WANG
- Publication Type:Journal Article
- MeSH:
Carcinoma;
genetics;
pathology;
Carcinoma, Papillary;
Cell Line, Tumor;
Cell Proliferation;
Humans;
Peptides;
genetics;
Plasmids;
RNA Interference;
RNA, Messenger;
genetics;
RNA, Small Interfering;
genetics;
Real-Time Polymerase Chain Reaction;
Thyroid Cancer, Papillary;
Thyroid Neoplasms;
genetics;
pathology;
Transfection;
Trefoil Factor-3
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2015;29(13):1194-1198
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To explore the effect on proliferation and invasion of human papillary thyroid carcinoma K1 cells by application of small hairpin RNA (shRNA) silencing TFF3 gene expression.
METHOD:Using liposome transfection method, TFF3-shRNA targeting of TFF3 gene will be transient transfected to papillary thyroid carcinama K1 cells, inducing the corresponding gene silencing. The experiment set up blank control group (Con group), negative control group (ConNC group) and interference group (TFF3-shRNA group). The TFF3 protein and mRNA expression were evaluated by RT-PCR, Real time-PCR, immunocytochemistry and Western blot in K1 cells after TFF3-shRNA transfected. CCK-8 method and Scratch test were used to detect the change of proliferation ability and invasion ability respectively.
RESULT:(1) The recombinant plasmid Ca # HSH018037-4-HIVmU6 carrying TFF3-shRNA transfected K1 cells successfully. (2) RT-PCR and Real time-PCR detected the expression of TFF3 mRNA, which was 0.38 ± 0.11 times as many as the blank control group (P < 0.01) after TFF3 gene silenced. But the negative control group was 1.082 times of blank control group (P > 0.05). (3) Western blot show that after TFF3 gene silence induced TFF3 protein expression levels have decreased 59.5% (P < 0.01), The difference was statistically significant compared with the blank control group. (4) Cell scratch detects K1 cell invasion ability. The invasion ability of K1 cells in interference group (TFF3-shRNA group) reduced. The scratch width significantly decreased 57.1% than blank control group (P < 0.01). (5) CCK-8 kit detect cell proliferation ability. K1 cells grow significantly slower in the interference group (TFF3-shRNA group) than the blank control group through the analysis of the growth curve (P < 0.01). In the interference group (TFF3-shRNA group) proliferation inhibition rate of K1 cells at 6 h, 12 h, 24 h and 36 h, 48 h are 16.6%, 26.6%, 33.6%, 33.8%, 35.0% respectively. Compared with negative control group, proliferation ability of K1 cell decreased significantly.
CONCLUSION:Silenced TFF3 gene can cause the degradation of mRNA, reduce the protein translation , and inhibit the invasion and proliferation ability of K1 cell.
