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Background and purpose:Gastric cancer is the most common malignant tumor of digestive tract, and the possibility of postoperative recurrence and metastasis is higher. Our previous studies showed that protein tyrosine phosphatase1B (PTP1B) gene is closely correlated with tumor size, lymph node metastasis and TNM stage of gastric cancer. The purpose of the present study was to explore the effect ofPTP1B gene on cell proliferation and migration of gastric cancer cell lines.Methods:Short hairpin RNA (shRNA) sequence targetingPTP1B gene and PTP1B cDNA were transfected into MKN28 and MKN45 cells, respectively. The expression of PTP1B mRNA and its protein in MKN28 and MKN45 cells were detected by quantitative real-time PCR (qRT-PCR) and Western blot, respectively. The effect of PTP1B on cell proliferation and migration was respectively assessed by cell counting kit-8 (CCK-8) assay, Transwell migration assay and wound healing assay.Results:Compared with blank and negative controls, the expressions of PTP1B mRNA and protein in MKN28 cells were successfully suppressed after the cells were transfected with shRNA (P<0.05). As CCK-8 test showed, the proliferation of MKN28 cells was successfully restrained at 48, 72 and 96 h after transfected with shRNA compared with blank control and negative control (P<0.05). Transwell and wound healing test were performed after PTP1B expression was interfered by shRNA. The result demonstrated that migration of MKN28 cells was signiifcantly inhibited (P<0.05). On the contrary, the expressions of PTP1B mRNA and protein in MKN45 cells were obviously enhanced after the cells were transfected with PTP1B cDNA. And the proliferation and migration of cells were significantly increased.Conclusion:PTP1B gene is an important enchancer for the proliferation and migration of gastric cancer.
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Objective To investigate the clinical value of anus-preserving operation for ultra low rectal cancer.Methods The clinical data of 226 patients with ultra low rectal cancer who were admitted to the Yijishan Hospital between January 2009 and September 2013 were retrospectively analyzed.All the patients were divided into the anus-preserving group (117 patients underwent traditional or modified Dixon operation) and the control group (109 patients underwent Miles operation).The operation time,volume of intraoperative blood loss,number of lymph node dissection,recovery time of intestinal function,postoperative short-term amplications,local recurrence and distal metastasis of tumors,survival rate and quality of life in the 2 groups were analyzed.The measurement data with normal distribution were presented as (x) ± s and analyzed using t test,count data were analyzed using Pearson chi-square test.The survival curve was drawn by Kaplan-Meier method,and the survival rate was analyzed using the Log-rank test.The patients were followed up by regular out-patient examination and telephone interview up to September 2014.Results The traditional Dixon operation was performed on 108 patients,modified Dixon operation on 9 patients and Miles operation on 109 patients in the control group.The operation time and volume of blood loss were (117 ± 12) minutes and (110 ± 51) mL in the anus-preserving group,which were significantly different from (122 ± 8) minutes and (155 ± 44) mL in the control group (t =3.80,7.09,P ＜ 0.05).The number of lymph node dissected and time of intestinal function recovery were 13 ±4 and (2.8 ± 0.8)days in the anus-preserving group and 13 ±4 and (2.7 ± 0.7)days in the control group,respectively,with no significant difference (t =0.90,0.83,P ＞ 0.05).Among the 184 patients receiving postoperative chemotherapy,19 patients terminated the chemotherapy due to serious response to chemotherapy,17 patients received radiotherapy after chemotherapy,9 patients received interventional treatment for liver after chemotherapy and 42 patients didn't receive the chemotherapy.The incidence of perianal eczema were 15.38％ (18/117) in the anus-preserving group and 3.67％ (4/109) in the control group,with a significant difference between the 2 groups (x2=8.81,P ＜0.05).The cases of anastomotic leakage,intra-abdominal bleeding,intra-abdominal infection,postoperative urinary retention,infection of incision and incision dehiscence were 4,3,5,10,7 and 9 in the anus-preserving group,which were not significantly different from 0,2,4,11,8 and 5 in the control group (x2=1.86,0.14,0.05,0.16,0.19,0.94,P ＞ 0.05).One-hundred and ninety patients were followed up for a median time of 34.5 months (range,12.0-57.0 months).The rate of local tumor recurrence,rate of distant metastasis and 1-and 3-year survival rate were 8.55％ (10/117),5.98％ (7/117),94.8％ and 76.2％ in the anus-preserving group and 8.26％ (9/109),5.50％ (6/109),95.7％ and 76.1％ in the control group,with no significant difference between the 2 groups (x2 =0.01,0.02,0.08,0.00,P ＞ 0.05).The score of life quality was 66 ± 14 in the anus-preserving group,which was significantly higher than 49 ± 11 in the control group (t =10.13,P ＜ 0.05).Conclusion The anus-preserving operation for ultra low rectal cancer is safe and feasible based on strictly grasping operative indications,meanwhile,it can improve the postoperative life quality of patients.
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It has been demonstrated that the combination of abnomal expression of gastrin with its receptor can promote cell proliferation and inhibit apoptosis in colorectal neoplasms via signal transduction pathways,which is related with the abnormal expression of miRNA.Researches show that there are multiple intersections between the molecule mechanisms of miRNA and the signaling pathway of gastrin in promoting colorectal cancer cell proliferation.It may exist a gastrin-miRNA-target gene signaling transduction pathway.Knowing the mechanism of the signaling pathway and controlling the related factors will open another avenue for the gene targeted therapy of colorectal neoplasms.
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Objective To investigate the effects and mechanisms of extracellular-signal regulated protein kinase-motogenactived protein kinase(ERK-MAPK)signaling pathway in gastrin-induced cell proliferation and apoptosis of colorectal cancer cells.Methods HT-29 cells were incubated in different media,and then were divided into the control group,gastrin group,proglumide group and gastrin + proglumide group.No reagent was added in the control group,and other groups were dealed with reagent in different concentrations.The changes of proliferation of the HT-29 cells were detected by MTT assay,and the optimal concentration of gastrin and proglumide were determined.The changes of proliferation index and apoptotic rates of HT-29 cells were detected by cell cytometry.The mRNA expressions of gastrin receptor/cholecystokinin-B receptor(CCK-BR),ERK1/2 and K-ras were detected by RT-PCR.The protein of ERK1/2,K-ras protein and phosphorylation levels were detected by Western blot.All data were analyzed by analysis of variance and SNK-q test.Results The proliferation of HT-29 was stimulated by gastrin when the concentration of the gastrin was 6.25-100.00 mg/L,and the optimal concentration of gastrin was 25.00 mg/L(F =31.36,P ＜ 0.05).Proglumide had no obvious effects on the proliferation of HT-29 cells,while it significantly inhibited the proliferation of HT-29 cells stimulated by gastrin when the concentration of proglumide was 8.00-128.00 mg/L,and the optimal concentration was 32.00 mg/L(F =24.31,P ＜ 0.05).The proliferation index of the gastrin(25.00 mg/L)group was 37.5 ％ ± 5.2％,which was significantly higher than 27.7％ ± 5.0％ of the control group and 27.3％ ± 5.8％ of the gastrin(25.00 mg/L)+ proglumide(32.00 mg/L)group(q =4.56,4.75,P ＜ 0.05).The apoptotic index of the gastrin(25.00 mg/L)group was 1.9％ ± 0.4％,which was significantly lower than 2.5％ ± 0.4％ of the control group and 2.4％ ± 0.3％ of the gastrin(25.00 mg/L)+ proglumide(32.00 mg/L)group(q =4.23,4.06,P＜0.05).The mRNA expression of CCK-BR was detected in the HT-29 cells.The levels of phosphorylated ERK1/2 protein and phosphorylated K-ras protein were 0.43％ ± 0.04％ and 0.45％ ± 0.06％,which were significantly higher than 0.32％ ± 0.02％ and 0.31％ ± 0.05 ％ of the control group(q =7.78,4.95,P ＜ 0.05),and they were also higher than 0.36％ ± 0.01％ and 0.35 ％ ± 0.04％ of the gastrin(25.00 mg/L)+ proglumide(32.00 mg/L)group(q =5.72,4.08,P ＜0.05).There were no significant differences in the mRNA and protein expressions of ERK1/2 and K-ras among the control group,gastrin(25.00 mg/L)group,proglumide(32.00 mg/L)group and gastrin (25.00 mg/L)+ proglumide(32.00 mg/L)group(F =0.52,0.72,0.78,0.28,P ＞0.05).Conclusion Gastrin could stimulate the proliferation of HT-29 cells and inhibit their apoptosis by upregulate the phosphorylation levels of ERK and K-ras through the Ras→Raf→ MEK1/2→ ERK1/2 pathway,while the effect can be restrained by gastrin receptor antagonist proglumide.
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Although patients with intestinal polyposis are not usual in clinical at present,it is still difficult to make early diagnosis because of the lack of specific clinical manifestations So clinicians should know the etiology of intestinal polyposis progress,master the methods of early diagnosis,select the appropriate treatment and do the follow-up regularly,which may help improving the prognosis of patients and improving the quality of survival.Early detection,early diagnosis and early treatment can largely avoid possible malignant polyps cancerous changing,which may not only have a significant effect on the quality of patients'life,but also have great significance to alleviate the burden of the patient's family.
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It has been reported that the high expression of gastrins and their receptors promotes some colorectal neoplasms cells proliferation and inhibit apoptosis.MicroRNAs (miRNAs) are closely related to gene expression regulation in colorectal neoplasms.Researches show that miRNAs have multiple intersections in the regulatory network of colorectal neoplasms with the signal transduction pathway of gatrins which controls the proliferation of colorectal neoplasms.Gastrins perhaps control the proliferation of colorectal neoplasms cells through miRNA signal transduction pathways.These findings have greatly expanded the understanding of the pathoge nesis of colorectal neoplasms and will provide new ideas and methods for the diagnosis and treatment of colorectal neoplasms.
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Objective To investigate the effects of somatostatin on the apoptosis of colorectal cancer cells. Methods The expression of somatostatin mRNA in colorectal cancer tissues from 79 patients who had been admired to Yijishan Hospital from January 2004 to October 2006 was detected by nested RT-PCR. The apoptotic index of colorectal cancer cells was detected by TUNEL, and the protein expressions of somatostatin, Fas, FasL, caspase-3 and caspase-8 in colorectal cancer tissues were detected by immunohistochemistry. All data were analyzed by chi-square test, q test and Spearman rank correlation coefficient. Results There was a positive correlation between the mRNA and protein expression of somatostatin (r = 0.98, P < 0.05). The mRNA and protein expression of somatostatin in poorly and moderately differentiated colorectal cancers were significantly lower than that in well differentiated colorectal cancers (χ~2 = 10.78, 11.24, 5.27, 5.24, P < 0.05). The positive expression rates of mRNA and protein of somatostatin in papillary adenocarcinoma were significantly higher than those in mucinous adenocarcinoma and signet ring cell carcinoma and undifferentiated carcinoma (χ~2= 6.56, 6.99, 5.44, 7.39, P < 0.05). The mRNA and protein expression of somatostatin in colorectal cancer in Dukes A and B were significantly higher than that in Dukes C and D (χ~2 =5.17, 4.06, P <0.05). The apoptotic index in high or moderate somatostatin expression group was significantly higher than that in low somatostain expression group (q = 5.66, 4.21, P < 0.05), and the positive expression rates of Fas, caspase-8 and caspase-3 in high or moderate somatostatin expression group were significantly higher than those in low somatostafin expression group (χ~2= 5.48, 5.62, 6.89, 4.32, 4.19, 3.91, P <0.05). Conclusion Somatostatin plays an important role in the regulation of cell apoptosis in colorectal cancer, and the mechanism may be related to the aberrant expression of Fas/FasL.
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AIM:To construct eukaryotic expression vector expressing double shRNA sections targeting Survivin gene.METHODS: Eukaryotic expression vector expressing double shRNA sections targeting Survivin gene were designed and chemically synthesized.They were directionally inserted into plasmid pGenesil-1 with respectively U6 promoter and termination code,the common green fluorescence protein(EGFP) gene and Neo gene. In this way,the vector of pGenesil-1 shRNA containing 2 sections of Survivin shRNA were constructed and they were transfected into the pancreatic cancer cell Bx-PC3.Transfection was detected by fluorescence microscope.The inhibition expression of Survivin mRNA was measured by RT-PCR.RESULTS: HE1 and HE2 plasmids were identified by the biocatalyst cut which confirmed the exactitude and were analyzed by the sequence analysis which verified the perfect clone plasmid inserted by them.CONCLUSION: A eukaryotic expression vector of double short hairpin RNA for Survivin gene is successfully constructed.The pancreatic cancer cells Bx-PC3 succeed to be transfected and expression of Survivin mRNA is inhibited obviously.
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Objective To explore the correlation between the expression of somatostatin (SS), gastrin (GAS), apoptosis index (AI),and p53 gene in colonic carcinoma. Methods The expression of GAS, SS, p53 and apoptosis cell were detected by immunohistochemistry (streptavidin-biotin-pero xidase complex, SABC) and in situ apoptosis detecting technic (TUNEL) . Results AI is higher in SS high and middle expression cases than in low expression cases(q=5.06,q=3.95,P