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 investigate the most appropriate culture time with the action of EGF in colon cancer stem cells enrichment by suspension culture.MethodsDLD-1 cells were cultured in serum-free medium containing 20 ng/mL EGF to generate spheroid cells. The time gradient was set to 10 d, 20 d, 30 d and 40 d, the cell proportion of CD133+, CD44+ and CD133+CD44+ were confirmed by flow cytometery. The ability of self-renewal was detected by the sphere forming assay and the limited dilution assay, and the in vitro tumorigenicity of the cells was detected by the colony formation assay.ResultsIn the 30 d group, the proportion of CD133+ and CD133+ CD44+ cells were significantly higher than those in the other groups (allP<0.05), the CD44+ cell was higher than that in the 20 d group (P<0.05), but there was no significant difference with the other two groups (P>0.05). The results of the limited dilution assay and the colony formation assay, the number of spheres in the 30 d or 40 d group was the highest among the 4 groups, and there was no statistical difference between the 30 d group and 40 d group (P>0.05), with statistically significant difference between the 30 d, 10 d and 20 d groups (all P<0.05). The results of the sphere forming assay and the self-renewal ability of 30 d group was significantly higher compared with other groups (all P< 0.05).ConclusionThe cancer stem cells could be enriched more efficiently by suspension culture using 20 ng/mL EGF for 30 days.
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.
Objective To evaluate the feasibility of sentinel lymph node (SLN) mapping after 99Tcm sulfur colloid (99Tcm-sc) and carbon nanoparticles injection in patients with colon cancer. Methods Forty patients with colon cancer underwent complete mesocolic excision between August 2015 and July 2016 at Qingdao Central Hospital were considered for prospective inclusion. Before resection, SLN mapping was performed with injection of 99Tcm-sc and carbon nanopar-ticles, then all dissected lymph nodes were detected by pathological examination. Results A total of 660 cases of lymph nodes were found in the 40 patients (average of 16.5 cases per patient). Of them, 88 nodes (average of 2.2 cases per patient) were identified as SLN in 36 of 40 patients, with a successful detection rate of 90.0% (36/40). The diagnostic accuracy, sensitivity, and false-negative rate were 87.5% (35/40), 96.2% (25/26), and 3.8% (1/26) respectively. Conclusion 99Tcm-sc and carbon nanoparticles suspension injection for mapping SLN is a feasiblely diagnostic method for predicting local lymph node metastasis in the patient with colon cancer.
Objective To investigate the expression of Bloom syndrome (BLM) helicase and tumor infiltrating dendritic cells (TIDC) in colon cancer tissues and their relationship with the prognosis of patients after surgery. Methods Onehundred and sixty-eight patients with colon cancer who underwent surgical resection in our hospital from June 2014 to August 2016 were selected as the research objects. The specimens of surgically resected colon cancer tissues and adjacent tissues archived by the pathology department were obtained, and the expression of BLM helicase and TIDC density were detected by immunohistochemistry. Pearson was used to analyze the correlation between BLM helicase expression and TIDC density, and the relationship between the expression of BLM helicase and TIDC density and the clinicopathological features of colon cancer was analyzed by using χ2 test or Wilcoxon rank test. The influencing factors of postoperative survival of patients with colon cancer were analyzed by Cox proportional hazards regression model. Results The relative expression of BLM helicase in colon cancer tissues was higher than that in adjacent tissues (1.49±0.33 vs. 1.02±0.17), while the TIDC density was lower than that in adjacent tissues [(9.53±2.36)% vs. (12.36±2.37)%], the differences were statistically significant (P<0.05). Pearson correlation analysis showed that there was a negative correlation between the expression of BLM helicase and TIDC density (r=–0.588, P<0.05). The expression of BLM helicase and TIDC density were correlated with tumor differentiation, clinical stage and lymph node metastasis (P<0.05). That is, those with high expression of BLM helicase and low density of TIDC had low degree of tumor differentiation, late clinical grade, and higher ratio of lymph node metastasis. Sixty-three cases died (37.5%) during the follow-up period (16–60 months, medium follow up 45 months). Log-rank analysis showed that the 5-year cumulative survival rate of the BLM helicase-low expression group was higher than that of the high expression group, and that of the TIDC-low density group was lower than that of the high density group (P<0.05). Cox regression analysis showed that the high expression of BLM helicase, low density of TIDC, low degree of tumor differentiation, late stage and lymph node metastasis were risk factors affecting the postoperative survival of patients with colon cancer (P<0.05). Conclusion The abnormal expression of BLM helicase and TIDC density in colon cancer tissues are related to the degree of differentiation and lymph node metastasis, which are risk factors affecting the long-term survival of patients with 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.
ObjectiveCombined with long non-coding RNA (lncRNA) to find a regression model that can be used to predict the survival rate of patients with colon cancer before operation.MethodsThe clinical information and gene expression information of patients with colon cancer were downloaded by using TCGA database. The differentially expressed lncRNAs in tumor and paracancerous tissues were screened out, and then combined with the clinical information of patients to construct Cox proportional hazard regression model.ResultsA total of 26 kinds of lncRNAs with statistical difference in gene expression between paracancerous tissues and tumor tissues were selected (P<0.05). Through repeated screening and comparison of prediction efficiency, the prediction model was finally selected, which was constructed by patients’ age, M stage, N stage, and three kinds of lncRNAs (ZFAS1, SNHG25, and SNHG7) gene expression level: age [HR=4.00, 95%CI: (1.48, 10.84), P=0.006], M stage [HR=3.96, 95%CI: (2.23, 7.04), P<0.001], N stage [HR=1.87, 95%CI: (1.24, 2.84), P=0.003], ZFAS1 gene expression level [HR=0.60, 95%CI: (0.41, 0.86), P=0.006], SNHG25 gene expression level [HR=0.85, 95%CI: (0.73, 1.00), P=0.045], and SNHG7 gene expression level [HR=2.32, 95%CI: (1.53, 3.52), P<0.001] were all independent risk factors for postoperative survival of patients with colon cancer. The area under the ROC curves for predicting 1, 3, and 5-year overall survival were 0.802, 0.828, and 0.771, respectiely, which had a good prediction ability.ConclusionThe predictive model constructed by the combination of ZFAS1, SNHG25, SNHG7 genes expression level with M stage, N stage, and age can better predict the overall survival rate of patients before operation, which can effectively guide clinical decision-making and choose the most suitable treatment method for patients.
ObjectiveTo investigate the feasibility and safety of the double cavity casing negative pressure drainage by inside and outside of the intestine in the primary resection and anastomosis of left colon cancer combined with acute obstruction. MethodsEighty-one cases of left colon cancer combined with acute obstruction who underwent surgeries in our hospital from January 2009 to December 2012 were collected prospectively, and were divided into one-stage surgery group (n=41) and control group (n=40). Cases of one-stage surgery group received double cavity casing negative pressure drainage by inside and outside of the intestine in the primary resection and anastomosis, and cases of control group underwent two-stage surgeries. Comparison of operation time, blood loss, time of anal exhaust after operation, hospital stay, hospital expense, and incidence of complication between the 2 groups was performed. ResultsThere were no significant difference in the operation time[(166±19) minutes vs. (173±23) minutes], blood loss[(253±42) mL vs. (273±50) mL], and time of anal exhaust after operation[(3.24±0.73) days vs. (3.50±0.95) days]beeween one-stage surgery group and control group, but hospital stay[(15.1±2.3) days vs. (23.1±4.1) days]and hospital expense[(3.70±0.68) ×105 yuan vs. (5.77±0.95) ×105 yuan]of one-stage surgery group were lower than those of control group (P<0.05). In addition, there were no significant difference in the incidences of wound infection[7.3% (3/41) vs. 10.0% (4/40)], intraabdominal infection[4.9% (2/41) vs. 10.0% (4/10)], pulmonary infection[12.2% (5/41) vs. 15.0% (6/40)], and anastomotic leakage[2.4% (1/41) vs. 5.0% (2/40)]beeween one-stage surgery group and control group (P>0.05). All of the cases were followed up for 1-36 months, and the median time were 22 months. There were no significant difference in the mortality[0 (0/41) vs. 2.5% (1/40)], recurrence rate[2.4% (1/41) vs. 5.0% (2/40)], and metastasis rate[7.3% (3/41) vs. 10.0% (4/40)]beeween one-stage surgery group and control group too (P>0.05). ConclusionIn the case of negative pressure drainage of double cavity casing, underwent decompression of the small bowel, and irrigation of colon, the primary resection and anastomosis of left colon cancer combined with acute obstruction was safe and feasible.
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 explore the relevant risk factors for postoperative delirium (POD) in elderly patients undergoing radical colon cancer surgery, and provide a basis for formulating postoperative prevention and treatment measures for POD. MethodsA total of 128 elderly patients diagnosed with colon cancer and underwent radical colon cancer surgery at Xindu District People’s Hospital in Chengdu from January 2018 to December 2021 were included as the study subjects. Patients were divided into two groups according to the score of Delirium Assessment Scale (4AT Scale). The basic data, main perioperative clinical data and laboratory indicators of the two groups were collected, and univariate and logistic regression analysis were carried out to determine the potential risk factors of POD in elderly patients with colon cancer after radical operation. ResultsAccording to the results of the 4AT scale score, a total score of ≥4 points was used as the threshold for determining patient POD. Among 128 patients, there were 29 patients (22.66%) with POD and 99 patients (77.34%) without POD. ① General data comparison: There was no significant difference between the two groups in gender, body mass index, years of education, hypertension, diabetes, smoking history and drinking history (P>0.05), but there was significant difference in age, preoperative mini-mental state examination (MMSE) score and American Society of Anesthesiologists (ASA) grade (P<0.05). ② Comparison of main clinical data during the perioperative period: There was no statistically significant difference between the two groups of patients in ICU treatment, nonsteroidal anti-inflammatory drug treatment, visual analogue scale, and intraoperative hypotension (P>0.05), but there was a statistically significant difference in operative time, anesthesia time, intraoperative blood loss, and dexmedetomidine treatment (P<0.05). ③ Comparison of preoperative laboratory indicators: There was no statistically significant difference between the two groups of patients in terms of hemoglobin, serum albumin, white blood cell count, prognostic nutritional index, neutrophil/lymphocyte ratio, D-dimer, and albumin to fibrinogen ratio (P>0.05). ④ The results of logistic regression analysis showed that low preoperative MMSE score [OR=0.397, 95%CI (0.234, 0.673)], long surgical time [OR=1.159, 95%CI (1.059, 1.267) ], and long anesthesia time [OR=1.138, 95%CI (1.057, 1.226) ] were independent risk factors for the occurrence of POD in elderly colon cancer patients undergoing radical surgery. ConclusionPreoperative MMSE score, operative time, and anesthesia time are closely related to the occurrence of POD in elderly colon cancer radical surgery, worth implementing key perioperative management in clinical practice to prevent and manage POD.