Construction of a recombinant eukaryotic vector of human intestinal trefoil factor and its expression in 293-T cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone human intestinal trefoil factor (hITF/hTFF3) gene into an eukaryotic expression vector for its expression in eukaryotic cells.</p><p><b>METHODS</b>The total RNA was extracted from normal human colon mucosa, and transcribed into cDNAs using RT-PCR. hTFF3 gene was amplified by PCR and ligated into pGEMT vector by TA cloning method. After sequencing, the hTFF3 gene was transfered into the eukaryotic expression vector pCMV5-myc. The recombinant vector was transfected into 293-T cells, and the expression of the recombinant protein was detected by Western blotting.</p><p><b>RESULTS AND CONCLUSION</b>hTFF3 gene was successfully cloned from normal human colon mucosa. The vector pCMV5-myc-hTFF3 was reconstructed, and in 293-T cells transfected with the vector, hTFF3 expression was detected by Western blotting.</p>

Molecular forms of TFF1 in normal gastric mucosa and its relationship with canceration / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the molecular forms of TFF1 in normal gastric mucosa and its expression in normal, gastric carcinoma, atypical hyperplasia, and intestinalized gastric mucosa.</p><p><b>METHODS</b>The molecular forms of TFF1 in normal gastric mucosa was observed by western blotting. The expression of TFF1 in normal, gastric carcinoma, atypical hyperplasia, and intestinalization gastric mucosa was assayed immunohistochemically.</p><p><b>RESULTS</b>TFF1 existed in normal gastric mucosa in forms of monomer, dimer and 21-kD TFF1 complex, with the last being the richest. TFF1 was expressed mainly in the epithelial cytoplasm of the mucosa in the gastric body and antrum, especially around the nucleus, and the closer to the lumen, the higher the expression. TFF1 expression in the tissues adjacent to gastric carcinoma was higher than that in normal gastric mucosa (P<0.001), and the expression in gastric adenocarcinoma was positively correlated to differentiation of adenocarcinoma. No TFF1 was expressed in poorly differentiated adenocarcinoma. The expression of TFF1 in moderate and well differentiated adenocarcinoma was a little lower than that in normal mucosa (P>0.05). The gastric mucosa with atypical hyperplasia had significantly higher TFF1 expression than normal gastric mucosa (P<0.001), and TFF1 was not detected in intestinalized gastric mucosa. There was no significant difference in TFF1 expression between gastric mucosa around the intestinalized tissues and normal gastric mucosa (P>0.05).</p><p><b>CONCLUSIONS</b>TFF1 plays an important part in protection and restitution of the gastric mucosa, and TFF1 may be related to suppression and differentiation of carcinoma.</p>