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[The effect of TFF3 on the proliferation and migration of papillary thyroid carcinoma K1 cell]
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.
Insights
Silencing the TFF3 gene in papillary thyroid carcinoma K1 cells significantly inhibits cell proliferation and invasion. This study demonstrates TFF3 gene silencing
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer.
- The trefoil factor 3 (TFF3) gene is implicated in various cancers, but its role in PTC proliferation and invasion requires further elucidation.
Purpose of the Study:
- To investigate the impact of silencing the TFF3 gene on the proliferation and invasion capabilities of human papillary thyroid carcinoma K1 cells.
- To understand the molecular mechanisms underlying TFF3's role in PTC progression.
Main Methods:
- Human papillary thyroid carcinoma K1 cells were transfected with TFF3-specific small hairpin RNA (shRNA) using a liposome-based method to achieve gene silencing.
- Gene silencing was confirmed by evaluating TFF3 mRNA and protein expression using RT-PCR, Real-time PCR, immunocytochemistry, and Western blot.
- Cell proliferation was assessed using the CCK-8 assay, and cell invasion was evaluated through the scratch test.
Main Results:
- Successful silencing of the TFF3 gene was achieved, leading to a significant reduction in both TFF3 mRNA (0.38 ± 0.11 fold) and protein expression (59.5% decrease) compared to control groups.
- Silencing TFF3 significantly inhibited K1 cell invasion, with a 57.1% decrease in scratch width.
- Cell proliferation was markedly reduced in TFF3-silenced cells, with inhibition rates reaching up to 35.0% at 48 hours.
Conclusions:
- TFF3 gene silencing effectively reduces TFF3 mRNA and protein levels in K1 cells.
- Downregulation of TFF3 inhibits the proliferation and invasion of human papillary thyroid carcinoma K1 cells.
- These findings suggest that TFF3 is a potential therapeutic target for papillary thyroid carcinoma.
