ObjectiveTo investigate the effect of Wnt5a derived from tumor-associated fibroblasts （CAFs） on the migration and invasion of gastric cancer cells. MethodsThe differentially expressed genes Wnt5a between CAFs and normal gastric fibroblasts (NGFs) in gastric cancer tissues and their corresponding normal gastric tissues using the GEO database GSE194261 dataset were screened. Immunohistochemical method was used to detect the expression of Wnt5a protein in tissue samples of clinical gastric cancer patients, and the relationship between Wnt5a protein expression and clinicopathological features of gastric cancer was analyzed. CAFs and NGFs were extracted from fresh surgical specimens of gastric cancer patients, and the expression of Wnt5a in CAFs was detected by real-time fluorescence quantitative-polymerase chain reaction and Western blot experiment. Transwell invasion and migration experiment was used to observe the effects of CAFs, inhibition of Wnt5a expression in CAFs and different concentrations of recombinant Wnt5a protein on the migration and invasion ability of gastric cancer MGC-803 and MKN-28 cell lines in vitro. ResultsThrough the screening of GEO database GSE194261 data set, it was found that Wnt5a was more expressed in CAFs than NGFs (P<0.05). Immunohistochemical results showed that the expression of Wnt5a protein in gastric cancer tissues was significantly stronger than that in normal gastric tissues (P<0.05), and the expression of Wnt5a protein was related to T stage of tumor (χ2=5.035, P<0.05), but not related to gender, age, degree of tumor differentiation, lymph node metastasis, vascular invasion and nerve invasion (P>0.05). Inhibiting Wnt5a derived from CAFs could inhibit the invasion and migration of gastric cancer cells. By stimulating gastric cancer cells with different concentrations of human recombinant Wnt5a protein, it was found that when the concentration of human recombinant Wnt5a protein was greater than 100 ng/mL, the invasion and migration abilities of MGC-803 and MKN-28 gastric cancer cells were significantly increased (P<0.05). ConclusionWnt5a is highly expressed in CAFs derived from the interstitial tissue of gastric cancer, which is related to the invasion depth of gastric cancer and can promote the invasion and migration of gastric cancer cells.
ObjectiveTo assess value of preoperative clinical data and enhanced CT imaging features in predic-tion of microvascular invasion (MVI) and early recurrence (recurrence in one year) after curative resection for hepatoce-llular carcinoma (HCC). MethodsA retrospective analysis was conducted for 150 patients with HCC who underwent curative tumor resection in West China Hospital of Sichuan University from April 2014 to May 2015. The roles of preoperative CT characteristics and clinical data on MVI and early recurrence after curative tumor resection were evaluated by univariate and multivariate analyses. Resultscompared with HCC with no MVI and no early recurrence after curative resection, univariate analysis results showed that HCC with MVI and early recurrence had larger tumor size (P=0.002, P=0.005), a higher proportion of non-smooth tumor margin (P<0.001, P<0.001), and tumor multifocality (P=0.005, P=0.038), HCC with MVI had a higher proportion of incomplete tumor capsule (P=0.032), HCC with early recurrence had a higher proportion of incomplete and absence tumor capsules (P=0.038) and a faster washout on portal venous phase-the percentage attenuation ratio on the portal venous phase (P=0.049) and relative washout ratio on the portal venous phase (P=0.020) were higher. A multivariate logistic regression analysis results showed that non-smooth tumor margin (OR=7.075, P<0.001; OR=4.125, P<0.001) and tumor multifocality (OR=3.290, P=0.008; OR=2.354, P=0.047) were the independent predictors for MVI and early recurrence after curative tumor resection, HCC with early recurrence also had a faster washout on the portal venous phase (OR=1.023, P=0.017). ConclusionNon-smooth tumor margin and tumor multifocality are independent risk factors for MVI and early recurrence after curative tumor resection, and HCC with early recurrence has a faster washout on portal venous phase. Preoperative enhanced CT imaging could predict MVI and early recurrence after curative tumor resection and CT imaging findings are helpful to choose reasonable treatment and predict prognosis.
ObjectiveTo investigate the influence of heat shock protein A2 (HSPA2) on the biological behavior of pancreatic adenocarcinoma cells and its mechanism. MethodsThe expressions of HSPA2 were determined in the human pancreatic adenocarcinoma cell lines (PANC-1, BxPC-3, and AsPC-1) using the Western blot. Subsequently, the cells with the lowest and highest HSPA2 expressions among these three lines were selected for conducting overexpression and knockdown experiments targeting HSPA2, respectively. The cellular proliferation, cell clonogenesis, migration, and invasion capabilities were assessed using MTT, clonogenic assay, and Transwell assay, respectively. Additionally, the impact of HSPA2 on the expression of key markers of epithelial-mesenchymal transition (EMT) was examined using the Western blot. The potential target molecules of HSPA2 were identified through immunoprecipitation assay and mass spectrometry. The rescue experiments further explored the regulatory relation between the HSPA2 and its target molecules. The influence of HSPA2 on pancreatic adenocarcinoma growth was investigated through establishment of xenograft tumor model in nude mice. ResultsThe HSPA2 exhibited the lowest expression in the PANC-1 cells and the highest expression in the AsPC-1 cells among the three cell lines. Subsequent functional studies demonstrated that the overexpression of HSPA2 in the PANC-1 cells markedly promoted proliferation, cell clonogenesis, migration, and invasion, while the knockdown of HSPA2 expression in the AsPC-1 cells markedly inhibited these processes. The Western blot analysis further showed that the HSPA2 overexpression downregulated E-cadherin expression and upregulated N-cadherin and Vimentin expressiones, whereas the HSPA2 knockdown produced opposite effects. The rescue experiments indicated that the HSPA2 promoted the EMT in pancreatic adenocarcinoma cells by upregulating Yes associated protein (YAP). The subcutaneous xenograft tumor experiments in the nude mice showed that the HSPA2 knockdown inhibited tumor growth. ConclusionThe results of this study suggest that HSPA2 promotes EMT via upregulating YAP, which facilitates proliferation, migration, and invasion of pancreatic adenocarcinoma cells.
Objective To assess the impact of adjuvant chemotherapy (ACH) on the survival of patients with pT3 urothelial carcinoma of the bladder (UCB) after radical cystectomy (RC). Methods We retrospectively analyzed the clinical and follow-up data of 223 UCB patients who underwent RC between January 2005 and June 2015. None of the patients received neoadjuvant chemotherapy. Of all the patients, 75 (33.6%) were diagnosed as pT3 cancer (including 32 pT3a and 43 pT3b patients). The follow-up data were up to June 2015. Kaplan-Meier method with log-rank test was used to estimate and compare overall survival (OS) and cancer-specific survival (CSS) between groups. Multivariate Cox proportional hazard models were used to identify predictors of OS and CSS. Results The short-term total effective rate of gemcitabine and cisplatin assisted chemotherapy in the treatment of pT3 UCB was 60.0%. Five-year OS rate (47.9%vs. 43.3%) and CSS rate (57.4%vs. 57.6%) were similar in the pT3a and pT3b groups (P=0.682 and 0.796, respectively). In pT3 patients, adjuvant chemotherapy was an independent predictor for OS (P=0.032). On multivariate analysis, according to the pT3 sub-stage, ACH was significantly associated with improved OS [hazard ratio (HR) =0.37; 95% confidence interval (CI) (0.15, 0.68),P=0.006] and CSS [HR=0.34, 95%CI (0.12, 0.86),P=0.022] in the pT3b group only. Conclusion Because pT3b cancer is characterized by macroscopic peri-vesical tissue invasion, patients may obtain an OS benefit from the administration of adjuvant chemotherapy.
Objective To evaluate the clinical value of ureteroscope in cholelithiasis treated by laparoscopic surgery. Methods The clinical data of 36 patients admitted because of hepatolithus with ureteroscope combination in laparoscopic surgery from February 2007 to September 2009 in Guidong People’s Hospital of Guangxi were analyzed retrospectively. Results In 33 cases, stones were removed once by ureteroscope in laparoscopic surgery with residual stones (in 3 cases residual stone were removed secondarily through T tube) and the other 3 cases were transferred to laparotomy forcedly due to bleeding of biliary duct and vessels of porta hepatis and tearing of bile duct. During operation, blood loss was 30-280 (94.51±54.70) ml; operation time was 110-260 (147.22±48.45) min; recovery time of bowel movement was 1-3 (2.03±0.76) d; postoperative hospitalization time was 6-13 (7.12±1.65) d (some discharged with T tube); the time of patients of T tubes pulled out was 28-45 (38.92±6.52) d. Bile leakage happened in 1 case and infection of biliary tract in 1 case, no complications such as biliary stricture or bile duct bleeding were found after operation. Conclusions Treatment of intrahepatic bile duct or a single extra-hepatic sand-like stones with ureteroscopy usage in laparoscopic surgery is feasible and less invasive. It is a minimally invasive treatment for intra- or extra-hepatic stones due to rapidly postoperative rehabilitation.
ObjectiveTo explore the value of multi-slice CT angiography (MSCTA) in peripancreatic vascular invasion of pancreatic carcinoma. MethodsThirty-eight patients with pancreatic carcinoma were detected by MSCTA technology before operation. The peripancreatic vascular invasion of pancreatic carcinoma was evaluated by multi-planar reconstruction (MPR) and maximum intensity projection (MIP) combined with axial image, and compared with the surgical results. ResultsThe MSCTA results showed that there were 12 patients (31.6%) with vascular invasion in 38 patients with pancreatic carcinoma, and the surgical results showed that there were 16 patients (42.1%) with vascular invasion. There was a b fit goodness of two results (kappa=0.665, P=0.000). The sensibility and specificity of MSCTA was 68.8% (11/16) and 95.5% (21/22), respectively. ConclusionsMSCTA technology has a high correct rate in evaluation of peripancreatic vessel encroached by pancreatic carcinoma, the MSCTA result has a b consistency to the surgical result. It has a value of clinical application in evaluation of peripancreatic vessel encroached by pancreatic carcinoma.
Objective To evaluate the short-term effectiveness of minimally invasive total hip arthroplasty (THA) by direct anterior approach (DAA). Methods Between January and August 2014, THA was performed on 48 patients (60 hips) by DAA (group A), and on 72 patients (92 hips) by posterolateral approach (group B). There was no significant difference in gender, age, etiology, course, and preoperative visual analogue scale (VAS), Harris hip score (HHS), and hip range of motion (ROM) between 2 groups (P>0.05). The operation time, intraoperative blood loss, postoperative drainage, hospitalization time, incision healing, and complications were recorded and compared. The acetabular abduction and anteversion were measured on the X-ray films; prosthesis loosening was observed. The VAS score, HHS score, and hip ROM were used to evaluate the hip function. Results The operation time and intraoperative blood loss of group A were significantly higher than those of group B, and the hospitalization time was significantly lower than group B (P<0.05), but no significant difference was found in postoperative drainage between 2 groups (t=0.71,P=0.46). The patients were followed up 2-2.5 years (mean, 2.2 years) in group A, and 2-2.5 years (mean, 2.1 years) in group B. In group A, 3 cases had lateral femoral cutaneous nerve traction injury and 1 case had swelling and exudate, and primary healing of incision was obtained in the other cases of group A and all cases of group B. No periprosthetic joint infection occurred in the others of groups A and B except 1 case of group A at 2 months after operation, and infection was controlled after debridement, irrigation, and intravenous infusion of Vancomycin for 1 month. The X-ray films showed good position of prosthesis and no obvious radiolucent line or prosthesis loosening. There was no significant differences in acetabular abduction and anteversion between groups A and B at last follow-up (P>0.05). The VAS score, HHS score, and hip ROM at 3 months and last follow-up were significantly better than preoperative ones in 2 groups (P<0.05), but no significant difference was found between at 3 months and last follow-up (P>0.05). The VAS score, HHS score, and hip ROM in group A were significantly better than those in group B at 3 months postoperatively (P<0.05). At last follow-up, the hip ROM in group A was significantly better than that in group B (P<0.05), and there was no significant difference in VAS and HHS scores between group A and group B (P>0.05). Conclusion The short-term effectiveness of minimally invasive THA by DAA is satisfactory, with the advantage of little trauma, short hospital stay, and rapid postoperative recovery.
ObjectiveTo investigate effect of Notch pathway regulating by inhibiting expression of forkhead box protein A1 (FOXA1) on proliferation and invasion of colon cancer SW480 cells. MethodsThe colon cancer tissues and their corresponding paracancerous tissues of 45 patients with colon cancer admitted to the First Affiliated Hospital of Henan University of Science and Technology from June 2019 to February 2021 were selected. The immunohistochemistry and real-time fluorescent quantitative PCR (qRT-PCR) methods were used to detect the expressions of FOXA1 protein and mRNA in the tissues, respectively. In addition, SW480 cells were divided into control group (untreated), shRNA-NC group (transfected with shRNA-NC), sh-FOXA1 group (transfected with sh-FOXA1), sh-FOXA1+sodium valproate group (Add 8 mmol/L Notch pathway activator sodium valproate after transfection with sh-FOXA1). Then the qRT-PCR, MTT, clone formation test, and Transwell methods were used to detect the expressions of FOXA1 mRNA, proliferation, clonogenic ability, invasion and migration of cells in each group. Western blot method was used to detect the proliferation (c-Myc, cyclinD1), invasion and migration [matrix metalloproteinase (MMP）9, MMP2], epithelial-mesenchymal transition (Vimentin, N-cadherin, E-cadherin) and Notch pathway (Notch-1, Hes-1) related protein expressions of cells in each group. Results① In the clinical cases, the expression levels of FOXA1 protein and mRNA in the colon cancer tissues were higher than those in the corresponding paracancerous tissues (protein: 0.085±0.028 vs. 0.034±0.010, t=11.036, P<0.001; mRNA: 1.62±0.34 vs. 1.00±0.09, t=11.671, P<0.001). ② In the cell experiment, compared with the control group and shRNA-NC group, the cell survival rate, and numbers of cloned cells, invasion and migrating cells were significantly reduced (P<0.05), correspondingly, the related proteins expression levels of c-Myc, cyclinD1, MMP9, MMP2, Vimentin, N-cadherin, Notch-1, Hes-1 were significantly reduced (P<0.05) and the protein expression level of E-cadherin was significantly increased (P<0.05) in the sh-FOXA1 group, which were reversed after adding the Notch pathway activator sodium valproate (P<0.05). ConclusionFOXA1 highly expresses in colon cancer tissues and colon cancer cells and it might promote the proliferation, invasion and migration of SW480 cells by activating the Notch pathway.
ObjectiveTo evaluate therapeutical effects of Huaier granule combined with transcatheter arterial chemoembolization (TACE) following radical resection of primary hepatocellular carcinoma with microvascular invasion. MethodsThe clinical data of 45 cases of primary hepatocellular carcinoma with microvascular invasion underwent Huaier granule combined with transcatheter arterial chemoembolization (TACE) following radical resection from June 2010 to June 2013 in Liaoning Cancer Hospital were retrospectively analyzed. These patients were divided into Huaier granule plus TACE treatment group (20 cases) and simple TACE treatment group (25 cases) according to the postoperative treatment of the patients. The immune function (CD4+/CD8+ ratio and IL-2 level), 1and 3-year tumor recurrence rates and 3-year cumulative survival rate were compared between two groups after operation. Result① The CD4+/CD8+ ratio and IL-2 level had no significant differences between the 2 groups before operation (P > 0.05), which in the Huaier granule plus TACE treatment group were significantly higher than those in the simple TACE treatment group (P < 0.05) on month 3, 6, and 12 after operation.② 1and 3-year tumor recurrence rates in the Huaier granule plus TACE treatment group were significantly lower than those in the simple TACE treatment group[15% (3/20) versus 48% (12/25), P < 0.05; 45% (9/20) versus 80% (20/25), P < 0.05]. ③ The 3-year cumulative survival rate was 75% and 68% in the Huaier granule plus TACE treatment group and the simple TACE treatment group, respectively. The survival curve analysis showed that the 3-year survival rate had a decreased trend, which in the Huaier granule plus TACE treatment group was slightly higher than that in the simple TACE treatment group, but the difference had no statistical significance between the 2 groups (P > 0.05). ConclusionsAlthough the results of this study fails to confirm that Huaier granule plus TACE treatment for primary hepatocellular carcinoma with microvascular invasion following radical resection could significantly improve the 3-year cumulative survival rate, it could effectively decrease the recurrence rate. It is needed larger sample size to further explore in future research.
ObjectiveTo investigate the relationship between DDX46 genes and invasion and migration of esophageal squamous cell carcinoma cells. MethodsHuman esophageal squamous cell carcinoma cells TE-1 were transfected by fluorescent marker shRNA lentivirus (shDDX46 group), and an empty vector was transfected as a control (shCtrl group). The expression rate of green fluorescent protein under the microscope was used to evaluate the cell transfection efficiency. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blotting (WB) detected the knockdown efficiency of the target gene at the mRNA and protein expression levels. Wound healing, invasion assay and migration assay detected the changes of invasion and metastasis ability. Classical pathway analysis was used to explore signaling pathway changes and the possible mechanism of DDX46 in the invasion and metastasis was explored by detecting fibronectin expression. ResultsDDX46 gene at mRNA and protein levels was significantly inhibited after lentiviral transfection. Wound healing showed that after 8 h the cell mobility of TE-1 cells decreased significantly (P=0.001). Invasion assay showed that after 24 h the average cell metastasis rate of TE-1 cells was lower in the shDDX46 group than that in the shCtrl group (P<0.001). The cell metastasis rate in the shDDX46 group corresponding to observation points in the transwell assay was lower than that in the shCtrl group (P<0.001) after 24 h culture. The results of the classical pathway analysis showed that the integrin signaling pathway activity was inhibited, further exploration of the mechanism of action found that the expression of fibronectin associated with cell adhesion was decreased. ConclusionDDX46 gene is related to the invasion and migration ability of esophageal squamous cell carcinoma cells. Knockdown of DDX46 genes may reduce cell adhesion by downregulating the integrin pathway signaling.