Objective To explore the relationship between Epstein-Barr virus (EBV) infection with serum human kissin-1 (KISS-1) and matrix metalloproteinase 2 (MMP2) levels and prognosis in patients with colon cancer. Methods A total of 86 colon cancer patients who were hospitalized in our hospital from April 2015 to April 2016 were selected as the colon cancer group; in the same period, 84 cases of physical examination person in our hospital were selected as the control group. Real-time fluorescent quantitative PCR (qRT-PCR) was used to test colon cancer patients for EBV DNA, and divided the patients into EBV DNA negative group and EBV DNA positive group according to the test results. Enzyme-linked immunosorbent (ELISA) method was used to detect serum KISS-1 and MMP2 levels. Pearson method was used to analyze the correlation between serum KISS-1 and MMP2 levels in patients with colon cancer infected with EBV. The survival curve was drawn by Kaplan-Meier method, and the relationship between EBV infection and prognosis of colon cancer patients was analyzed by log-rank test. Multivariate Cox regression analysis were used to analyze the factors affecting the prognosis of colon cancer patients. Results Compared with the control group, the positive rate of EBV DNA in the colon cancer group was higher (χ2=21.854, P<0.001). The EBV DNA positive rate of patients with lymph node metastasis, TNM stage Ⅲ–Ⅳ, tumor low differentiation and tumor infiltration T3–T4 was higher than those without lymph node metastasis, TNM stage Ⅰ–Ⅱ, tumor high/medium differentiation and tumor infiltration T1–T2 (P<0.05). Compared with the EBV DNA negative group, the serum KISS-1 level of the EBV DNA positive group decreased, and the MMP2 level increased (P<0.001). There was a negative correlation between serum KISS-1 and MMP2 levels in colon cancer patients with EBV infection (r=–0.510, P<0.001). The 5-year survival rates of colon cancer patients in the EBV DNA-negative group and the EBV DNA-positive group were 52.94% (27/51) and 14.29% (5/35), respectively, the difference between the two groups was statistically significant (χ2=13.274, P<0.001). EBV infection, MMP2 high expression, and lymph node metastasis were independent risk factors affecting the prognosis of colon cancer patients (P<0.05), and KISS-1 low expression was a protective factor affecting the prognosis of colon cancer patients (P<0.05). Conclusions EBV infection is closely related to the progression and prognosis of colon cancer. The down-regulation of KISS-1 and the up-regulation of MMP2 may be related to EBV infection.
Objective To investigate the effects of recombinant adenovirus-mediated co-transfection of carcinoembryonic antigen (CEA) gene and erythropoietin (EPO) gene on promoting hematopoietic stem cells directly producing erythrocyte vaccine against colon cancer. Methods The expression adenovirus vectors carrying CEA and EPO or green fluorescent protein (GFP) gene were constructed respectively, and recombinant adenovirus carrying CEA, EPO or GFP were packaged and produced respectively. The bone marrow-derived mesenchymal stem cells (MSCs) of mice were isolated and cultured in vitro by anti-CD117 magnetic bead separation, and were transfected with CEA (CEA group), EPO (EPO group) or GFP (blank vector group), co-transfected with CEA and EPO (CEA-EPO group). The expressionsof CEA and EPO gene and its protein after transfection in supernatant fluid of culture were detected by realtime-PCR and Western blot method in each group. We had checked and obtained the vaccine with co-transfection of CEA gene and EPO gene by cell red line marker antibody CD71 and GPA, then we carried on experiments with the vaccine in vitro and in vivo. There were 4 groups in our trail: blank vector group, CEA group, EPO group, and CEA-EPO group. Results We had successfully gathered the hematopoietic stem cells, flow cytometry analysis result showed that there were significant differences before and after purification for positive selected samples (P<0.05). The expressions of double genes (CEA-EPO gene) and protein showed CEA-EPO gene were successfully transfected into the hematopoietic stem cells. We had confirmed erythrocyte vaccine with co-transfection of CEA and EPO gene by antibody CD71 and GPA with flow cytometry. The monocytes cytotoxicity on colon cancer cell line CT26 showed that lysis of target cells of CEA-EPO group were higher than those of other 3 groups when in proportion of 40∶1 (P<0.05). In the experimentation of neoplasma format, the volume of tumor and mortality were smaller or lower, but survival time was longer of CEA-EPO group in2 weeks after treatment (P<0.05). Conclusions The erythrocyte vaccine with co-transfection of CEA gene and EPO gene has efficient anti-tumor effects on colon cancer. Not only can promote hematopoietic stem cell directly producing erythrocyte vaccine, but also can produce tumor antigen vaccine against colon cancer.
ObjectiveTo investigate the effects of overexpression of alpha/beta hydrolase domain-containing protein 5 (ABHD5) on the invasion and migration of human colon cancer cell line HCT116 and the pathway of adenosine monophosphate-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR).MethodsThe expression of ABHD5 in colon cancer tissues and its relationship with clinicopathological features was analyzed by UALCAN database. HCT116 cells were cultured in vitro and transfected with ABHD5 recombinant plasmid, then they were divided into control group, negative transfection group and ABHD5 transfection group. Real time quantitative PCR (qRT-PCR) was used to detect the expression of ABHD5 mRNA in HCT116 cells. The proliferation of HCT116 cells was detected by CCK-8 method. Transwell assay was used to detect the invasion and migration of HCT116 cells. The expression of matrix metalloprotein 9 (MMP-9), E-cadherin, Snail, and AMPK/mTOR pathway proteins p-AMPK, AMPK, p-mTOR and mTOR were detected by Western blot.ResultsThe results of the UALCAN showed that compared with normal colon tissues, the expression of ABHD5 mRNA in colon cancer tissues was decreased (P<0.05), and which in the adenocarcinoma and the N1 stage was lower than that of the mucinous adenocarcinoma (P<0.05) and N0 stage (P<0.05), respectively. Compared with the control group and the negative transfection group, the expression of ABHD5 mRNA in the ABHD5 transfection group was increased (P<0.05), the proliferation inhibition rate of HCT116 cells in the ABHD5 transfection group was increased (P<0.05), the numbers of migration and invasion cells in the ABHD5 transfection group were decreased (P<0.05), the expressions of MMP-9, Snail, p-mTOR and mTOR were reduced, and the expressions of E-cadherin, p-AMPK and AMPK were increased (P<0.05).ConclusionsThe overexpression of ABHD5 can inhibit the invasion and migration of colon cancer HCT116 cells, activate AMPK, and inhibit the expression of mTOR. It suggests that ABHD5 may play a role in inhibiting colon cancer by affecting AMPK/mTOR pathway.
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 compare the short- and long-term effects of emergency surgery (ES) and self-expanding metal stent (SEMS) in treatment of malignant left-sided colonic obstruction.MethodsThe patients with malignant left-sided colonic obstruction who met the inclusion and exclusion criteria in the Third Affiliated Hospital of Soochow University from October 2010 to October 2020 were retrospectively collected and divided into ES group (n=43) and SEMS group (n=22). The baseline data, surgical data, postoperative data, and prognosis (overall survival and relapse free survival) were compared, and the risk factors of tumor recurrence after surgery were further analyzed by Cox proportional hazards regression model. ResultsIn this study, 65 cases of malignant left-sided colonic obstruction were included, including 43 cases in the ES group and 22 cases in the SEMS group. There were no statistical differences in the baseline data of the two groups (P>0.05). There were no significant differences in the incidence of postoperative complications [13.6% (3/22) vs. 23.3% (10/43), P=0.555], recurrence rate [40.9% (9/22) vs. 37.2% (16/43), P=0.772], and rate of receiving postoperative chemotherapy [68.2% (15/22) vs. 48.8% (21/43), P=0.138] between the SEMS group and ES group. Compared with the ES group, although the median hospitalization time was longer (20 d vs. 12 d, P=0.001), and the median hospitalization cost was higher (65 033 yuan vs. 40 045 yuan, P=0.001), the stoma rate of the SEMS group was lower [36.4% (8/22) vs. 88.4% (38/43), P=0.001], and the minimally invasive (laparoscopic) rate was higher [36.4% (8/22) vs. 7.0% (3/43), P=0.008]. There were no significant differences in the 4-year cumulative overall survival (46.9% vs. 48.4%, P=0.333) and 4-year cumulative relapse free survival (36.2% vs. 44.8%, P=0.724) between the SEMS group and ES group, but the overall survival of the SEMS group was better than that of the ES group for the patients with stage Ⅲ–Ⅳ (χ2=4.644, P=0.047). Multivariate analysis of Cox proportional hazards regression model showed that increased TNM stage increased the risk of postoperative tumor recurrence of patients with malignant left-sided colonic obstruction [HR=2.092, 95%CI (1.261, 3.469), P=0.004]. ConclusionsShort- and long-term effects of ES and SEMS in treatment of malignant left-sided colonic obstruction are equivalent. Although SEMS mode has a longer hospital stay and higher hospitalization costs, stoma rate is lower and laparoscopic surgery rate is higher. Overall survival of SEMS mode in treatment malignant left-sided colonic obstruction patients with stage Ⅲ–Ⅳ is better.
ObjectiveTo summarize the diagnosis and treatment of a primary gastric colon cancer, and to explore its safety and feasibility.MethodThe clinical data of a patient with gastric cancer and sigmoid colon cancer who admitted to The Affiliated Yantai Yuding Hospital of Qingdao University in October 2017 was analyzed.ResultsThe patient underwent laparoscopic radical gastrectomy plus π anastomosis and laparoscopic radical resection of colon cancer. The operation time was 330 min and the intraoperative blood loss was 120 mL. There were no complications such as stomach cramps and sputum after operation and he was successfully discharged on the 9th day after surgery. Postoperative pathological staging: gastric cancer (pT3N3M0, ⅢB) and sigmoid colon cancer (pT2N0M0, Ⅰ B).ConclusionsMultiple primary cancer of the simultaneous gastric colon should be diagnosed before operation. Laparoscopic minimally invasive treatment for gastric cancer with sigmoid colon cancer is safe and feasible, and can benefit patients.
ObjectiveTo explore the correlation of BMI and 25 hydroxyvitamin D3 level with colon cancer.MethodsA total of 100 cases who underwent colonoscopy and were excluded from bowel diseases at the physical examination center of the First Hospital of Qinhuangdao from March 2017 to October 2017 were retrospectively selected as the control group. A total of 100 patients who underwent colonoscopy at general surgery or physical examination center and were confirmed to have colon cancer by pathological examination were included in the colon cancer group. The height, weight and body mass index (BMI) were measured in the morning, and the level of 25 hydroxyvitamin D3 (25(OH)D3) was determined by fasting blood sampling.Results① There was no statistical significance in age and 25(OH)D3 level between the two groups (P>0.05), and BMI of the colon cancer group was significantly higher than that of the control group (P<0.05). ② The proportion of overweight and obesity in the colon cancer group was significantly higher than that in the control group (P<0.05), and the proportion of vitamin D deficiency was significantly higher as well (P<0.05). ③ Logistic regression analysis showed that the incidence of colon cancer in patients with vitamin D deficiency was 12.263 times higher than that in patients without vitamin D deficiency, and the incidence of colon cancer in patients with overweight and obesity was 2.215 times higher than that in patients with normal BMI, with statistically significant differences (P<0.05).ConclusionThe incidence of colon cancer in patients with vitamin D deficiency and those with BMI of overweight or obesity is significantly increased.
Objective To determine feasibility of texture analysis of CT images for the discrimination of hepatic epithelioid hemangioendothelioma (HEHE) and liver metastases of colon cancer. Methods CT images of 9 patients with 19 pathologically proved HEHEs and 18 patients with 38 liver metastases of colon cancer who received treatment in West China Hospital of Sichuan University from July 2012 to August 2016 were retrospectively analyzed. Results Thirty best texture parameters were automatically selected by the combination of Fisher coefficient (Fisher)+classification error probability combined with average correlation coefficients (PA)+mutual information (MI). The 30 texture parameters of arterial phase (AP) CT images were distributed in co-occurrence matrix (22 parameters), run-length matrix (1 parameter), histogram (4 parameters), gradient (1 parameter), and autoregressive model (2 parameters). The distribution of parameters in portal venous phase (PVP) were co-occurrence matrix (18 parameters), run-length matrix (2 parameters), histogram (7 parameters), gradient (2 parameters), and autoregressive model (1 parameter). In AP, the misclassification rates of raw data analysis (RDA)/K nearest neighbor classification (KNN), principal component analysis (PCA)/KNN, linear discriminant analysis (LDA)/KNN, and nonlinear discriminant analysis, and nonlinear discriminant analysis (NDA)/artificial neural network (ANN) was 38.60% (22/57), 42.11% (24/57), 8.77% (5/57), and 7.02% (4/57), respectively. In PVP, the misclassification rates of RDA/KNN, PCA/KNN, LDA/KNN, and NDA/ANN was 26.32% (15/57), 28.07% (16/57), 15.79% (9/57), and 10.53% (6/57), respectively. The misclassification rates of AP and PVP images had no statistical significance on the misclassification rates of RDA/KNN, PCA/KNN, LDA/KNN, and NDA/ANN between AP and PVP (P>0.05). Conclusion The texture analysis of CT images is feasible to identify HEHE and liver metastases of colon cancer.
ObjectiveTo investigate the regulatory role of DAB2IP in proliferation effect of colon cancer cells by salinomycin. MethodsCell counting kit 8 (CCK8) assay was used to detect median inhibitory concentration (IC50) of salinomycin on HT29 and SW480 cells. Colon cancer cells with stable knock-down of DAB2IP (HT29-shDAB2IP) and control cells (HT29-shcon) were constructed by lentivirus plasmid. And colon cancer cells with stable over-expression of DAB2IP (SW480-DAB2IP) and control cells (SW480-con) were constructed using pCI plasmid. The proliferation effect of salinomycin on stable knock-down or over-expression of DAB2IP by CCK8 assay in the colon cancer cell was identified. The colon cancer stem cells makers CD44, CD24, and CD133 were investigated using real-time PCR. ResultsThe salinomycin had obvious inhibitory effects on the proliferations of HT29 and SW480 cells, the IC50 value was 20.0 μmol/L and 10.0 μmol/L, respectively. The stable knock-down of DAB2IP could significantly enhance the inhibitory effect of salinomycin on the proliferation in HT29 cells (P<0.05) and stable over-expression of DAB2IP could significantly decrease the inhibitory effect of salinomycin on the proliferation in SW480 cells (P<0.05). Further the result of real-time PCR detection showed that the expressions of cancer stem cells markers CD44, D24, and CD133 were significantly increased after stable knock-down of DAB2IP in the HT29 cells (P<0.05), while the expressions were significantly decreased after stable over-expression of DAB2IP in the SW480 cells (P<0.05). ConclusionsFrom initial results of this study, salinomycin could suppress proliferation of colon cancer cells. DAB2IP might weaken proliferative inhibitory effect of salinomycin by inhibiting expressions of cancer cells stem in colon cancer cells.
Objective To investigate whether miRNA (miR)-34a mediates oxaliplatin (OXA) resistance of colon cancer cells by inhibiting macroautophagy via the transforming growth factor (TGF)-β/Smad4 pathway. Methods miR-34a expression levels were detected in colon cancer tissues and colon cancer cell lines by quantitative real-time polymerase chain reaction (qRT-PCR). Computational search, functional luciferase assay, and Western blotting method were used to demonstrate the downstream target of miR-34a in colon cancer cells. Cell viability was measured with cell counting kit-8. Apoptosis and macroautophagy of colon cancer cells were analyzed by flow cytometry and transmission electron microscopy, and expressions of Beclin1 and LC3Ⅱ protein were detected by Western blotting method. Results Expression of miR-34a was significantly reduced while expressions of TGF-β and Smad4 mRNA were increased in colon cancer patients treated with OXA-based chemotherapy. OXA treatment also resulted in decreased miR-34a expression levels and increased TGF-β and Smad4 expression levels in both parental cells and the OXA-resistant colon cancer cells. Activation of macroautophagy contributed to OXA resistance in colon cancer cells. Expression levels of Smad4 and miR-34a in colon cancer patients had a significant inverse correlation and overexpressing miR-34a inhibited macroautophagy activation by directly targeting Smad4 through the TGF-β/Smad4 pathway. OXA-induced downregulation of miR-34a and increased drug resistance by activating macroautophagy in colon cancer cells. Conclusion miR-34a mediates OXA resistance of colon cancer by inhibiting autophagy via the TGF-β/Smad4 pathway.