ObjectiveTo analyze the effects of miR-451a on the proliferation and apoptosis of human pancreatic cancer BxPc3 cells, and to explore its molecular mechanisms.MethodsThe liposome transfection mimics of miR-451a were established in the BxPc3 cells, which were used as the research objects, and different concentrations (25, 50, 100 and 200 μmol/L) of miR-451a and blank control group were set up respectively. The expression of miR-451a mRNA in the BxPc3 cells after the transfection was detected by the qRT-PCR method. The effects of miR-451a at different concentrations on the proliferation, cell clone number, cell cycle and apoptosis, and the expressions of the macrophage migration inhibitory factor (MIF), calcium binding protein 39 (CAB39), phosphorylated phosphatidylinositol-3-kinase (p-PI3K) and phosphorylated protein kinase B (p-AKT) proteins in the BxPc3 cells were detected by the MTT assay, plate cloning assay, flow cytometry, and Western blot, respectively.ResultsThe expressions of miR-451a mRNA in the transfected BxPc3 cells were significantly higher than in the blank control BxPc3 cells (P<0.050). The miR-451a could inhibit the proliferation of BxPc3 cells in a time- and concentration-dependent manner significantly (P<0.050), block the differentiation of BxPc3 cells in the G0/G1 phase, and induce the apoptosis with a concentration-dependent manner (P<0.050). The expressions of MIF, CAB39, p-PI3K, and p-AKT proteins in the BxPc3 cells were down-regulated with a concentration-dependent manner (P<0.050).ConclusionFrom results of this study, miR-451a could inhibit proliferation and induce apoptosis of BxPc3 cells in a concentration-dependent manner, and its mechanisms might be related to inhibition of PI3K/AKT signaling pathway.
ObjectiveTo investigate the expression of circular RNA mitochondrial fusin 2 (circ-MFN2) in pancreatic cancer and analyze its correlation with clinicopathological features and prognosis.MethodsThe expressions of circ-MFN2 miRNA in 55 cases of pancreatic cancer tissues and serum were detected by qRT-PCR, and analyzed the correlation between circ-MFN2 and clinicopathological factors of pancreatic cancer and prognosis. The sensitivity, specificity and accuracy of expression of circ-MFN2 miRNA in the diagnosis of pancreatic cancer were statistically analyzed. ROC curve was used to analyze its efficacy as a biomarker for early diagnosis of pancreatic cancer.ResultsCompared with paracancerous tissues of pancreatic cancer and serum of healthy control group, circ-MFN2 miRNA was highly expressed in pancreatic cancer tissues and serum, and the difference were all statistically significant (all P<0.05). Chi square test showed that the expression of circ-MFN2 miRNA in pancreatic cancer tissues was not related to age, gender, tumor size, pathological type, and tumor site (P>0.05), but was significantly related to CA19-9 level, TNM stage, tumor differentiation and lymph node metastasis (P<0.05). The sensitivity, specificity and accuracy of expression of circ-MFN2 miRNA for pancreatic cancer were 72.7%, 70.9% and 83.6% respectively, which were all higher than that of CA19-9 (54.5%, 50.9% and 52.7%, P<0.05). Kaplan-Meier analysis showed that the median survival time of pancreatic cancer patients with high expression of circ-MFN2 miRNA was significantly shorter than that of patients with low expression (9.1 months vs 22.3 months, P<0.05). The area under ROC curve of circ-MFN2 miRNA as a serum biomarker for the diagnosis of pancreatic cancer was 0.861 [95%CI (0.775, 1.157), P=0.000]. Cox multivariate analysis showed that the expression of cirC-MFN2 miRNA and lymph node metastasis were independent risk factors for the prognosis of pancreatic cancer patients.ConclusionsCirc-MFN2 miRNA is highly expressed in pancreatic cancer tissues, and it is related to the clinical characteristics and prognosis of patients. It is expected to be a new molecular marker to predict the prognosis of pancreatic cancer.
Objective To investigate the inhibition effect of silence of a disintegrin and metalloproteinase 17 (ADAM17) gene on proliferation and apoptosis of HT29 colon cancer cells and its possible mechanism. Methods HT29cells were divided into 3 groups: cells of interference group were transfected with recombinant lentivirus vector, cells of negative control group were transfected with negative recombinant lentivirus vector, and cells of blank control group were treated with PBS. The expression of ADAM17 mRNA was detected by real-time PCR, the expressions of ADAM17 protein, caspase3, protein kinase B (Akt), glycogen synthase kinase-3β (GSK3β), phospho-protein kinase B (P-Akt), phospho-glycogen synthase kinase-3β (P-GSK3β) protein were detected by Western blot method, the cell proliferation was detected by MTT assay, and the apoptosis rate was detected by Annexin V-FITC/PI cell death detection kit. Results Compared with the control group and the negative control group, the interference group was related to low expressions of ADAM17 mRNA and its protein, low optical density value at the same time point (24, 48, and 72 h), high apoptosis rate, high expression level of caspase3 protein, but low expression levels of P-Akt and P-GSK3β protein (P<0.05). Conclusion Silent ADAM17 gene could significantly induces apoptosis and inhibits the proliferation of HT29 cells, which maybe via inhibiting Akt/GSK3β signaling pathway.
ObjectiveTo investigate the expression level and the clinical significance of Traf-2 and Nck-interacting protein kinase (TNIK) in gastric cancer tissues.MethodsWe retrospectively collected 78 cases of gastric cancer and its adjacent tissues which were diagnosed by surgery and pathology, all the patients received surgery in the Department of General Surgery in Luoyang Central Hospital Affiliated to Zhengzhou University, from July 2014 to December 2017, as well as 42 cases of gastric ulcer tissues in the same period. The expression of TNIK protein was detected by using immunohistochemical method and Western blot method in 78 cases of human gastric cancer tissues, corresponding adjacent normal tissues, and 42 cases of gastric ulcer tissues, the relationship between expression of TNIK protein and clinicopathologial features of gastric cancer was explored.ResultsThe positive expression rate of TNIK protein in gastric cancer tissues was 66.67% (52/78), and adjacent cancer tissues showed low expression rate of 11.54% (9/78), while it was 21.43% (9/42) in gastric ulcer tissues. On the positive expression rate of TNIK protein, gastric cancer tissues compared with adjacent cancer tissues and gastric ulcer tissues, the difference was statistically significant (P<0.05), however, there was no statistically significant difference between adjacent cancer tissues and gastric ulcer tissues (P>0.05). Western blot method showed that the expression level of TNIK protein in gastric cancer tissues was all significantly higher than that in adjacent cancer tissues and gastric ulcer tissues, the differences were statistically significant (P<0.05), but the difference was not significant when compared the adjacent cancer tissues and gastric ulcer tissues (P=0.772). The expression of TNIK protein was irrelevent to sexual distinction, CEA value, tumor diameter, and pathological type (P>0.05), while it was closely related to patients’ age, TNM stage, differentiation grade, distant metastasis, and lymph node metastasis (P<0.05). The prognosis of patients with positive expression of TNIK protein was worse than that of patients with negative expression of TNIK protein (P<0.05).ConclusionTNIK protein is highly expressed in gastric cancer tissues, which is associated with poorly prognostic factors and poor prognosis, and it may be a therapeutic target in patients with gastric cancer.
Objective To explore the clinical value of preoperative neoadjuvant chemotherapy (NAC) combined with laparoscopic pancreatoduodenectomy (LPD) with multiple surgical approaches in the treatment of borderline resectable pancreatic head cancer. Methods The clinicopathologic data of 35 patients with critical resectable pancreatic head carcinoma admitted to the Department of Hepatobiliary and Pancreatic Surgery of Luoyang Central Hospital Affiliated to Zhengzhou University and the Department of Hepatobiliary and Pancreatic Surgery of the Fifth Affiliated Hospital of Zhengzhou University from January 2017 to June 2022 were retrospectively analyzed. All patients received NAC before operation (AG protocol). At the end of the course of treatment, according to the type of borderline resectable pancreatic cancer (BRPC) [venous invasion type (BRPC-V type) and arterial invasion type (BRPC-A type)], take the individualized surgical approach for LPD (BRPC-V type: inferior mesenteric vein approach; BRPC-A type: left posterior approach, medial uncinate process approach, anterior approach, or lower mesocolon approach). The intraoperative condition, R0/R1 resection rate, lymph node dissection, postoperative complications, average hospital stay, recovery, follow-up and survival were recorded. Results① Efficacy evaluation of NAC: 13 patients were partially relieved , 17 patients were stable and 5 patients were progressive after 4 weeks of treatment. Five progressive patients continued to receive comprehensive internal medicine treatment, and the remaining 30 patients underwent LPD. ② Intraoperative situation: LPD were successfully completed in 30 patients, 2 patients underwent extended pancreaticoduodenectomy combined with superior mesenteric vein (or) portal vein reconstruction among them. Among the 30 patients with LPD, there were 10 cases of inferior mesenteric vein approach, 10 cases of left posterior approach, 6 cases of medial uncinate process approach, 1 case of left posterior approach+medial uncinate process approach, 2 cases of anterior approach, and 1 case of inferior mesocolon approach. The mean operative time was (379.4±77.3) min, the intraoperative blood loss was (436.9±95.1) mL. ③ Postoperative situation: The incidence rate of postoperative surgery-related complications was 33.3% (10/30), including 4 cases of Clavien-Dindo grade Ⅰ [biliary fistula in 1 case (3.3%), A-grade pancreatic fistula in 1 case (3.3%), gastric draining dysfunction in 1 case (3.3%), diarrhea in 1 case (3.3%)], 5 cases of grade Ⅱ [pulmonary infection in 2 cases (6.6%), B-grade pancreatic fistula in 2 cases (6.6%), abdominal infection in 1 case (3.3%)], and 1 case of grade Ⅲ [gastroduodenal artery stump bleeding (3.3%)]. Among the 10 patients with complications, 9 cases recovered after symptomatic treatment, and 1 case died, with a fatality rate of 3.3% (1/30). The mean postoperative hospital stay was (17.3±5.5) days. ④ Excision rate and pathological results: R0 resection rate was 90.0% (9/10) in 10 patients with BR-PV type LPD, and R1 resection was performed in 1 patient. R0 resection rate was 75.0% (15/20) in 20 patients with BR-A type, and R1 resection was performed in 5 patients (2 patients with medial uncinate process approach; Left posterior approach in 2 cases; Submesocolon approach was used in 1 case). In 30 patients with LPD, the total R0 removal rate was 80.0% (24/30), the number of lymph nodes dissected was (11±5). Pathological type: There were 26 cases (86.7%) of ductal adenocarcinoma in 30 patients, 1 case of adeno-squamous carcinoma (3.3%), 1 case of mucinous carcinoma (3.3%), 2 cases of acinocytic cell carcinoma (6.7%). 23 cases (76.7%) of medium-high differentiation and 5 cases (16.6%) of low differentiation, two cases (6.7%) were undifferentiated. ⑤ Postoperative follow-up and survival: 30 patients were completely followed-up for 6-39 months, with a median follow-up time of 17 months. The median survival time of BRPC-V and BRPC-A patients was 24.0 months and 17.0 months, respectively. The overall survival rates of 30 patients at 1, 2 and 3 years after operation were 77.3%、46.5% and 13.7%, respectively. Conclusion The selection of preoperative NAC combined with individualized surgical approach for patients with borderline resectable pancreatic head cancer is beneficial to improve the radical resection rate and clinical therapeutic effect, and has good clinical application value.