INVESTIGATION OF THE RELATIONSHIP BETWEEN CELL PROLIFERATIVE ACTIVITY AND INVASION, METASTASIS AND PROGNOSIS OF GASTRIC CANCER

To evaluated Bromodeoxyuridine (BrdUrd) /DNA doubleparametric method in detection of gastric carcinoma and to analyze the relationships of cellular BrdUrd labeling indices(LI), G2/M-phase fraction(G2/MPE) and DNA content to the depth of invasion, lymphatic vessel invasion, lymphatic node metastasis, peritoneal dissemination and blood vessel invasion and prognosis. Methods: Fresh tumor samples from 60 cases were examined by BrdUrd/DNA doubleparametric flowcytometry. Propidium iodide(PI) was used as fluorescent probe for total cellular DNA and a monoclonal antibody against BrdUrd incorporated into DNA. Fluorescein-labeled goat anti-mouse antibody was used as second antibody. Results: The values of BrdUrd LI in patients with serosa invasion was significantly higher than those without serosa invasion (P<0.01); there was statistical significance in 5-year survival rate between the two groups (P<0.01). Both BrdUrd LI and G2/MPF values were significantly higher in patients with lymphatic vessel invasion than those without invasion (P<0.01); the patients with lymphatic vascular invasion carried a significantly poor prognosis (P<0.01). Both BrdUrd LI and G2/MPF values were significantly higher in patients with lymphatic node metastasis than those without metastasis (P<0.01), there was a statistical significance in 5 years survival between these 2 groups. The incidence of lymphatic node metastasis was significantly higher in aneuploid carcinoma (P<0.05), and the patients with aneuploid carried a significantly poor prognosis (P<0.05). Patients with peritoneal dissemination had a significantly worse prognosis (P<0.01). G2/MPF values were significantly higher in patients with blood vessel invasion than those without invasion (P<0.01). Conclusion: Cellular BrdUrd LI, G2/MPF and DNA content are related to depth of invasion, lymphatic vessel invasion, peritoneal dissemination, blood vessel invasion and prognosis of gastric carcinoma.

Decreasing Pin1 suppressed the properties of migratory and invasive in colorectal carcinoma SW620 cells

Objective:The aim of our study was to investigate the effect of Pin1 on the expression of MMP-2 and MMP-9 in human colorectal carcinoma SW620 cells. Methods: We constructed a eukaryotic expression vector of RNA interfering (shRNA) for Pin1 gene (pGenesil-1-Pin1), and then observed its expression in SW620 cells by Western blotting. The cells motility were tested by wound healing assay and Boyden chamber assay. The protein levels and activity of MMP-2 and MMP-9 were tested by Western blotting and Gelatin zymography in SW620 cells after transfected with pGenesil-1-PIN1. Results: pGenesil-1-PIN1 was successfully constructed, which was confirmed by sequencing. Silencing the Pin1 by RNAi significantly decreased the cells motility from 96.4±3.9 per field (×10 objective) to 52.7±4.4 per field (P<0.05, Student's t-test) for SW620 cells transfected with pGenesil-1-PIN1 (SW620/p-shRNA) in Boyden chamber assay, and reduced the MMP-2 and MMP-9 expressions and activity in SW620 cells. The protein relative levels of MMP-2 were 0.32±0.04 for SW620/p-shRNA, and 0.76±0.03 for SW620/p-Con; MMP-9 were 0.41±0.09 for SW620/p-shRNA, and 0.94±0.07 for SW620/p-Con (p<0.05). Conclusion: Inhibited Pin1 expression may contribute to the suppression of the invasive and metastatic capacity of colon cancer cells in vitro.