ABSTRACT
Objective To investigate the effects of TFF3 overexpression on the proliferation, migration, and invasion ability of colorectal cancer HT29 cells and the mechanisms involved in cancer metastasis. Methods HT29 cells were transfected with pIRES2-TFF3, and the expression levels of mRNAs and proteins related to TFF3 gene, TWIST1/TRIB3 signaling pathway, and epithelial-mesenchymal transition (EMT) were detected by qRT-PCR and Western blot. The proliferation, migration, and invasion ability of HT29 cells were detected by the CCK-8, cell scratch, and Transwell assays. Changes in cell morphology after TFF3 overexpression were observed through transmission electron microscopy. Results After the HT29 cells were transfected with pIRES2-TFF3, the expression levels of TFF3 mRNA and protein significantly increased (P<0.01); cell proliferation, migration, and invasion were significantly enhanced (P<0.01); and the expression of related genes, such as TWIST1, TRIB3, Vimentin, and Snail were significantly upregulated. By contrast, the expression of E-cadherin significantly decreased (P<0.05). Various changes in cell morphology were observed after TFF3 overexpression, such as decrease in cell junctions, loss of cilia, formation of pseudopodia, and increase in fusiform cells. Conclusion TFF3 overexpression may promote EMT in colorectal cancer cells through the Twist1/TRIB3 signaling pathway, increase their metastatic potential, and accelerate the malignant progression of colorectal cancer.
ABSTRACT
Objective MicroRNAs are differentially expressed in colorectal cancer tumor tissues and adjacent normal tissues, which play an important role in the development of colorectal cancer. This study aims to investigate the expression of microRNA-7-5p in colorectal cancer patients and its influence on the proliferation and apoptosis of colon cancer cell CaCo2. Methods The high-throughput microarray was used to screen differentially expressed microRNAs, and real-time quantitative PCR (RT-PCR) was used to detect the expression of microRNA-7-5p in 10 cases of colorectal cancer patients and corresponding adjacent normal tissues. Western blot was performed to detect the expression of intestinal trefoil factor (TFF3) in different tissues and CaCo2 cells after transfection with microRNA-7-5p. The expression of TFF3 in different tissues and CaCo2 cells transfected with microRNA-7-5p was detected. TUNEL combined with flow cytometry was used to detect the apoptosis of CaCo2 cells after transfection. The CCK-8 assay was used to detect the proliferation of CaCo2 cells after transfection. Results The relative expression of MicroRNA-7-5p in colorectal cancer tissue was 0.409 ± 0.095, which was significantly lower than that of normal tissue adjacent to cancer (1.000 ± 0.014), and the difference was statistically significant (P <0.01). The relative expression of TFF3 in colorectal cancer tissues was significantly higher than that in normal adjacent tissues (P <0.01). The relative expression of TFF3 in CaCo2 cells decreased in the overexpressed microRNA-7-5p of the control group (0.729 ± 0.041). The proliferation ability (0.930 ± 0.007) was significantly lower in the blank control group (0.990 ± 0.005) (P <0.01), and it could increase the proportion of early apoptotic cells (50.700 ± 0.989) and late apoptotic cells (40.525 ± 0.515). Conclusion MicroRNA-7-5p is lowly expressed in colorectal cancer and is associated with the occurrence and development of colorectal cancer. Up-regulated microRNA-7-5p inhibits the proliferation of colon cancer cell CaCo2 and promotes its apoptosis.
ABSTRACT
Gastric cancer is one of the most common cancers in the world. The aims of this study were to evaluate the association between polymorphisms in TFF gene family, TFF1, TFF2, and TFF3 and the risk of gastric cancer (GC) and GC subgroups in a Korean population via a case-control study. The eight polymorphisms in TFF gene family were identified by sequencing and genotyped with 377 GC patients and 396 controls by using TaqMan genotyping assay. The rs184432 TT genotype of TFF1 was significantly associated with a reduced risk of GC (odds ratio, [OR) = 0.45; 95% confidence interval, [CI] = 0.25-0.82; P = 0.009), more protective against diffuse-type GC (OR = 0.20; 95% CI = 0.05-0.89; P = 0.035) than GC (OR = 0.34; 95% CI = 0.14-0.82; P = 0.017) in subjects aged < 60 yr, and correlated with lymph node metastasis negative GC and diffuse-type GC (OR = 0.44; 95% CI = 0.23-0.86; P = 0.016 and OR = 0.20; 95% CI = 0.05-0.87; P = 0.031, respectively). In addition, a decreased risk of lymph node metastasis negative GC and diffuse-type GC was observed for rs225359 TT genotype of TFF1 (OR = 0.46, 95% CI = 0.24-0.88; P = 0.020 and OR = 0.21, 95% CI = 0.05-0.88; P = 0.033, respectively). These findings suggest that the rs184432 and rs225359 polymorphisms in TFF1 have protective effects for GC and contribute to the development of GC in Korean individuals.