ABSTRACT
Objective:To explore the inhibition of microRNA (miRNA, miR)-5590-3p on the expression of transforming growth factor beta typeⅡreceptor (TGFBR2) gene and its effect on the invasion and proliferation of gastric cancer cell line HS-746T.Methods:The gastric cancer cell line was cultured in vitro. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to analyze the expression of miR-5590-3p in gastric cancer tissues and cell lines. The miR-NC and miR-5590-3p mimic were transfected into gastric cancer cell line HS-746T by lipofection, and named as miR-NC group and miR-5590-3p group, respectively. qRT-PCR was used to measure transfection effects. Transwell assay and cell counting kit-8 (CCK-8) assay were used to detect cell invasion and proliferation after transfection. The bioinformatics software predicts and the dual luciferase reporter gene system validates the target gene of miR-5590-3p. qRT-PCR and western blot were used to measure the expression of TGFBR2 and its downstream proteins in the transfected cells. Results:The expression of miR-5590-3p in gastric cancer tissues was significantly lower than that in adjacent tissues ( P<0.01). The expression of miR-5590-3p in gastric cancer cell lines was significantly lower than that in normal gastric mucosal epithelial cells ( P<0.05), and lowest in HS-746T cells ( P<0.01). After transfection, the expression of miR-5590-3p in miR-5590-3p group (11.76±0.21) was significantly higher than that in miR-NC group (1.06±0.21), with statistically significant difference ( P<0.01). The number of invasive cells in miR-NC group and miR-5590-3p group were (101.20±15.47) and (26.53±6.53), respectively, and the invasion ability of miR-5590-3p group was significantly decreased ( P<0.01). Compared with the miR-NC group, the cell proliferation ability of the miR-5590-3p group was significantly decreased ( P<0.05). Bioinformatics software showed that the target gene for miR-5590-3p is TGFBR2. The dual luciferase reporter gene system confirmed that miR-5590-3p can target the TGFBR2 gene ( P<0.01). Western blot results showed that compared with miR-NC group, the expression of TGFBR2 in HS-746T cells in miR-5590-3p group was significantly decreased ( P<0.01), the expression of ZEB-1 and ZEB-2, and the expression of CDK1 and cyclin B proteins were decreased as well. Conclusions:miR-5590-3p can inhibit the invasion and proliferation of gastric cancer cell HS-746T by targeting and regulating the expression of TGFBR2 gene.
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Objective:To observe the expression of long non-coding RNA (lncRNA) PP7080 in gastric cancer tissues and cell lines, and clarify its effect on the migration and proliferation of gastric cancer cell MGC803 and its molecular mechanism.Methods:Real-time PCR was used to detect the expression of lncRNA PP7080 in gastric cancer tissues and adjacent tissues, gastric cancer cell lines and immortalized normal gastric mucosal epithelial cell lines. The gastric cancer cell line with the least expression was selected, and the expression plasmid of lncRNA PP7080 and the negative control plasmid were transfected into gastric cancer cells, respectively, and named as lncRNA PP7080 group and NC group. Real-time PCR to verify the effect of transfection. Cell scratch test and CCK-8 test were used to detect the regulation of lncRNA PP7080 on the migration and proliferation of gastric cancer cells.The statistical saftware SPSS 20.0 was used for statistical analysis, the measurement data of normal distribution were expressed as ( Mean± SD). Real-time PCR and Western blot were used to detect the expression of tumor metastasis suppressor gene 1 ( TMSG-1) in the transfected cells. The t test was used for comparison between groups. Results:The expression of lncRNA PP7080 in gastric cancer tissue was less than that in adjacent tissues [(0.85±0.34) vs (5.33 ± 0.76), P<0.01]. The expression of lncRNA PP7080 in gastric cancer cell lines is less than that of immortalized normal gastric mucosal epithelial cells ( P<0.05), and the least expression in MGC803 cells ( P<0.01). The expression of lncRNA PP7080 in the lncRNA PP7080 group was significantly higher than that in the NC group, and the difference was statistically significant ( P<0.01). The cell migration rates of NC group and lncRNA PP7080 group were (72.67±6.39)% and (26.45±6.63)%, respectively, and the cell migration ability of lncRNA PP7080 group was significantly reduced ( P<0.01). Compared with the NC group, the cell proliferation ability of the lncRNA PP7080 group was significantly reduced ( P<0.05). Compared with the NC group, the expression of TMSG-1 in MGC803 cells of the lncRNA PP7080 group was significantly reduced ( P<0.01). Conclusion:lncRNA PP7080 is lowly expressed in gastric cancer tissues and cell lines. lncRNA PP7080 can inhibit the migration and proliferation of gastric cancer cell MGC803 by promoting the expression of TMSG-1 gene.
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Objective:To explore the regulation of microRNA (miRNA, miR)-7856-5p on the expression of EPH receptor A3 (EPHA3) gene and its effect on the migration and proliferation of colorectal cancer cell line SW480.Methods:Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the expression of miR-7856-5p in colorectal cancer tissues and cell lines. The miR-7856-5p mimic and miR-NC were transfected into colorectal cancer cell line SW480 by lipofection, respectively, and defined as miR-7856-5p group and miR-NC group, respectively. Real-time PCR was used to detect transfection effects. Transwell assay and CCK-8 assay were used to detect cell migration and proliferation after transfection. The bioinformatics software predicts and the dual luciferase reporter gene system validates the target gene of miR-7856-5p. Real-time PCR and Western blot were used to detect the expression of EPHA3 in the transfected cells. The measurement data in accordamce with normal distribution were expressed as mean±standard deviation, t test was used for inter grap comparison, and single factor analysis of variance was used for multi group comparison. Results:The expression of miR-7856-5p in colorectal cancer tissues was significantly lower than that in adjacent tissues ( P<0.01). The expression of miR-7856-5p in colorectal cancer cell lines was significantly lower than that in normal intestinal mucosal epithelial cells ( P<0.05), and lowest in SW480 cells ( P<0.01). The expression of miR-7856-5p in miR-7856-5p group was significantly higher than that in miR-NC group, the difference was statistically significant [(9.49 ± 1.09) vs (1.06 ±0.18), P<0.01]. The number of migrated cells in miR-NC group and miR-7856-5p group were (125.70±14.05) and (42.01±8.98), respectively, and the migration ability of miR-7856-5p group was significantly decreased ( P<0.01). Compared with the miR-NC group, the cell proliferation ability of the miR-7856-5p group was significantly decreased ( P<0.05). Bioinformatics software showed that the target gene for miR-7856-5p was EPHA3. The dual luciferase reporter gene system confirmed that miR-7856-5p can target the EPHA3 gene ( P<0.05). Compared with the miR-NC group, the expression of EPHA3 in the SW480 cells of the miR-7856-5p group was significantly decreased ( P<0.05). Conclusion:miR-7856-5p can inhibit the migration and proliferation of colorectal cancer cell SW480 by regulating the expression of EPHA3.
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Objective To investigate risk factors related to postoperative death of patients with rectosigmoid junction tumor perforation.Methods The clinical data of 76 cases with rectosigmoid junction tumor perforation confirmed by laparotomy from January 2000 to October 2015 were collected.Results Of the 76 cases,17 patients died postoperatively,the mortality rate was 22％,the single factor analysis showed that age(x2 =4.649,P =0.031),duration of abdominal pain(x2 =8.218,P =0.016),severe heart and lung diseases(x2 =11.996,P =0.007),circulatory and renal function(x2 =10.360,P =0.016),serum albumin(x2 =7.252,P =0.027),white blood cell count(x2 =7.633,P =0.022),Perforation diameter (x2 =9.770,P =0.008),Geroge grade of intraperitoneal contamination (x2 =10.086,P =0.006) were related to postoperative death (P ＜ 0.05).Multivariate analysis showed that complicating severe heart and lung diseases,preexisted circulatory and renal dysfunction,white blood cell count ＜ 4 × 109/L,size ＞ 3 cm,intraperitoneal contamination larger than one quadrant were independent risk factors for postoperative death.Conclusion Risk factors related to postoperative death of rectosigmoid junction tumor perforation were preoperative important organ dysfunction and intraperitoneal infection.
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Objective To investigate the clinical efficacy of transabdominal preperitoneal laparoscopic hernia repair (TAPP). Methods Three trocars were introduced into the abdomen at the umbilicus and both sides 10 cm from the umbilicus, respectively. The parietal peritoneum was opened at 5 cm above the internal ring and near the external border of the umbilicus. The hernial sac was dissected and a patch overlying the defect was stapled with the pectineal ligament and the conjoined tendon. Results The laparoscopic operation was completed successfully in all the 50 cases. The operation time was 20~70 min (39.8?10.5 min). Postoperatively, there were 1 case of groin pain and 2 cases of urine retention. The duration of postoperative hospital stay was 1~4 days (mean, 2 days). Follow-up observations in the 50 cases for 1~11 months (mean, 7 months) found no recurrence. Conclusions Transabdominal preperitoneal laparoscopic hernia repair is a safe and effective tension-free hernioplasty.