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Article in Chinese | WPRIM | ID: wpr-706886


Objective: To observe the influence of TRPV6 gene silence on SW480 colon cancer cell biological behavior, change in the in-tracellular concentration of calcium, as well as influence of 1,25 (OH)2D3CaCl2and CuCl2on SD rat colonic neoplasm models. Method: SW480 colon cancer cells were infected using lentivirus particles. TRPV6 protein and mRNA expression was detected using immunohistochemical tests, Western blot, and PCR. Moreover, the proliferation, metastasis, and apoptosis of SW480 colon cancer cells were detected through MTT assay and metastasis and apoptosis experiments, and the concentration of Ca2+in SW480 colon cancer cells was measured using high-speed ionic imaging. The SD rat colon cancer model was established based on DMH, and were assigned into experimental group (DMH group, 15) and intervention group (DMH+1,25 (OH)2D3group, DMH+CuCl2group) and control group, 10 in each group. The SD rat colon cancer model is established based on DMH, given 1,25(OH)2D3(37.5 nmol/kg) and CuCl2(375 μmol/kg) separately as intervention. The occurrence of colonic neoplasms and glandular cancers in each group of rats was observed, and Western blot was employed for detection of the TRPV6 protein expression. Results: After the transfection of SW480 colon cancer cells by TRPV6-RNAi, the expression of TRPV6 mRNA and protein decreased, intracellular concentration of Ca2+decreased, proliferation and metastasis rate of SW480 colon cancer cells decreased, and apoptosis rate of these cells increased. The differences between the groups with intervention and the blank control group and negative control group showed statistical significance (P<0.05). The colon cancer occurrence rate in the control group was 0, while that of the DMH+1,25 (OH)2D3 group, DMH group, and DMH+CuCl2were 100%, 84.62%, and 33.33%, respectively. The TRPV6 protein expression was detected in all groups, while DMH+1,25(OH)2D3group was observed to exhibit the highest level of expression, followed by the DMH group, DMH+CuCl2group, and control group. The differences were of statistical significance (P<0.05). Conclusions: The proliferation and metastasis of SW480 colon cancer cells can be prohibited by lowering the concentration of Ca2+in the cells. Thus, the apoptosis of the cells can be induced. 1,25 (OH)2D3can help improve the expression of TRPV6 protein in experimental rat colon tissues and promote the formation of colon neoplasms. CuCl2can help lower the expression of TRPV6 protein in experimental rat colon tissues and prevent the formation of colon neoplasms.

Article in Chinese | WPRIM | ID: wpr-620782


Objective:To investigate the role of TRPV5 and TRPV6 in intracellular calcium regulation and biological behaviors of SW480 colon cancer cells. Methods:qRT-PCR, Western blot, and immunocytochemistry were applied to determine the mRNA and protein ex-pression levels of TRPV5 and TRPV6 in SW480 colon cancer cell line upon treatment with TRPV5 and TRPV6 agonist, 1-25(OH)2D3, and inhibitor, CuCl2. The change of intracellular Ca2+level was examined with a confocal laser scanning microscope. Scratch test, MTT, and TUNEL assays were used to analyze the cell migration, proliferation, and apoptosis, respectively. Results:As an agonist of TRPV5 and TRPV6, 1-25(OH)2D3 significantly up-regulated the mRNA and protein expression levels of TRPV5 and TRPV6 in SW480 cell lines. On the other hand, CuCl2, being an inhibitor of TRPV5 and TRPV6, effectively down-regulated the TRPV5 and TRPV6 mRNA and protein expres-sion levels (P<0.05). The intracellular calcium concentration in SW480 cell line significantly increased upon treatment with 1-25 (OH)2D3, and significantly decreased with CuCl2 treatment (P<0.05). 1-25(OH)2D3 promoted cell proliferation and migration, and inhibit-ed apoptosis of SW480 cell in a time-and dose-dependent manner (P<0.05). However, CuCl2 significantly repressed cell proliferation and migration and induced apoptosis (P<0.05). Conclusion: TRPV5 and TRPV6 can affect the biological behaviors of colon cancer SW480 cells by regulating intracellular Ca2+level.

Clinical Medicine of China ; (12): 1191-1194, 2008.
Article in Chinese | WPRIM | ID: wpr-397554


Objective To investigate the expression of gastrin, cyclooxygenase-2(COX-2) and Ki-67 in dif-ferent gastric mucosal lesions and to assess the role of gastrin and COX-2 in the sequence of gastric carcinogenesis and its relationship. Methods 96 samples with different gastric mucosal lesions were obtained by gastroscope. The expression of gastrin,COX-2 and Ki-67 was examined with immunohistochemical staining technique(EnVision). Re-sults Positive rates of gastrin expression in chronic superficial gastritis (CSG), chronic atrophic gastritis (CAG), intestinal metaplasia(IM),dysplasia(Dy) and gastric cancer(GC) were 58.8 % ,30. 0% ,57.9 % ,68.2% and 77.8% ,respectively, and there were significant differences between GC and CAG(P<0.05). Positive rates of COX-2 expression in CSG,CAG,IM,Dy and GC were 41.2 % ,45.0% ,68.4 % ,72.7% and 88.9% respectively, and there were significant differences between GC and CAG(P<0.05). Ki-67 proliferation index (PI) from CSG, CAG, IM, Dy to GC also had an increasing tendency. From CAG to GC,the expression of gastrin and COX-2 was positively correlated with PI (P<0.05). The expression between gastrin and COX-2 was correlated significantly in gastric pre-cancerous lesions (P<0.01). PI was increased significantly in group with co-expression of gastrin and COX-2. Con-clusion Gastrin expression is decreased in CAG. From CAG to GC, gastrin expression is increased gradually. COX-2 expression has an increasing tendency in the sequence of gastric cancerization. From CSG to GC, PI is gradually in-creased. Gastrin and COX-2 co-promote proliferation of gastric epithelial cells,and gastrin may take part in up-regu-lation of COX-2 expression in gastric precancerous.

China Pharmacy ; (12)2007.
Article in Chinese | WPRIM | ID: wpr-529652


OBJECTIVE:To investigate the efficacy of compound azintamide enteric-coated tablet plus mosapride citrate dispersible tablet for abdominal distention in patients with functional dyspepsia(FD).METHODS:A total of 80 patients with the chief complaint of abdominal distention were randomized to treatment group(Group A,n=40) to be treated with mosapride citrate dispersed tablets plus compound azintamide enteric-coated tablet,or control group in which the patients were treated bu mosapride citrate dispersed tablets(Group B,n=20) or compound azintamide enteric-coated tablets(Group C,n=20).Comp-ound azintamide enteric-coated tablets were administered orally tid(2 tablets) immediately after meal while the mosapride citrate dispersed tablets(5mg) were administered orally tid 30min before meal for 2 weeks.The main outcome measures were changes of symptom scores of abdominal distension,the total effective rates in improvement of abdominal distension,and drug side effects before and after treatment.RESULTS:After 1-week and 2-week treatment,the average index scores of abdominal distention in the treatment group(Group A) were significantly lower than in the control groups(Group B and Group C) (P