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.
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 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.
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.
Objective To investigate the role of long non-coding RNA metastasis-associated in colon cancer 1-antisense RNA （MACC1-AS1）in cisplatin resistant gastric cancer and its possible mechanism. Methods Human gastric cancer cell line BGC823 and cisplatin resistant gastric cancer cell line (BGC823/DDP) were selected as the research objects. BGC823/DDP cells were transfected and divided into negative control group (si-NC group, transfected with si-NC empty plasmid) and MACC1-AS1 gene silencing group (si-MACC1-AS1 group, transfected with si-MACC1-AS1 plasmid). The BGC823 cells were transfected and divided into positive control group (pcDNA-NC group, transfected with pcDNA-NC empty plasmid) and MACC1-AS1 gene overexpression group (pcDNA-MACC1-AS1 group, transfected with pcDNA-MACC1-AS1 plasmid). MTT was used to detect the inhibition and 50% inhibition concentration (IC50). Flow cytometry was used to detect apoptosis. Real-time fluorescence quantitative PCR was used to detect the mRNA expression levels of MACC1-AS1, B-lymphoma-2 gene (Bcl-2), Bcl-2 related X gene (Bax), mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), protein kinase B (AKT), and phosphorylated AKT (p-AKT). Western blot was used to detect the protein expression levels of Bax, Bcl-2, p-mTOR, mTOR, AKT, and p-AKT. Results The relative expression level of MACC1-AS1 mRNA in BGC823/DDP cells was higher than that in BGC823 gastric cancer cells (P<0.01). The relative expression level of MACC1-AS1 mRNA in the si-MACC1-AS1 group cells was lower than that in the si-NC group cells (P<0.01). The relative expression level of MACC1-AS1 mRNA in the pcDNA-MACC1-AS1 group cells was higher than that in the pcDNA-NC group cells (P<0.01). The cell growth inhibition rate and IC50 of the si-MACC1-AS1 group were higher than those of the si-NC group (P<0.01). The cell growth inhibition rate and IC50 of the pcDNA-MACC1-AS1 group were lower than those of the pcDNA-NC group (P<0.01). The mRNA and protein relative expression levels of Bcl-2, p-AKT/AKT and p-mTOR/mTOR in the pcDNA-MACC1-AS1 group were significantly higher than those in the pcDNA-NC group (P<0.01). The relative expression levels of Bax protein and mRNA in the pcDNA-MACC1-AS1 group were significantly lower than those in the pcDNA-NC group (P<0.01). The apoptosis rate of the pcDNA-MACC1-AS1 group was significantly lower than that of the pcDNA-NC group (P<0.01). The mRNA and protein relative expression levels of Bcl-2, p-AKT/AKT and p-mTOR/mTOR in the si-MACC1-AS1 group were significantly lower than those in the si-NC group (P<0.01). The relative expression levels of Bax protein and mRNA in the si-MACC1-AS1 group were significantly higher than those in the si-NC group (P<0.01). The apoptosis rate of the si-MACC1-AS1 group was significantly higher than that of the si-NC group (P<0.01). Conclusions MACC1-AS1 highly expresses in cisplatin resistant gastric cancer cells. Overexpression of MACC1-AS1 regulates AKT/mTOR pathway mediated apoptosis and enhances cisplatin resistance of gastric cancer cells.
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 relationship between preoperative hemoglobin, albumin, lymphocyte and platelet (HALP) score, and clinicopathologic features of colon cancer, and to analyze the predictive value of HALP score for postoperative liver metastasis. Methods The clinical data of 163 patients with colon cancer admitted to the 909th Hospital of Joint Logistic Support Force (Dongnan Hospital of Xiamen University) from January 2018 to December 2019 were retrospectively analyzed. According to the occurrence of postoperative liver metastasis, the patients were divided into metastatic group (n=35) and non-metastatic group (n=128). The correlation between preoperative HAPL score and clinicopathologic features of colon cancer was analyzed. The predictive value of HALP score for postoperative liver metastasis of colon cancer was analyzed by using receiver operating characteristic (ROC) curve. The risk factors of liver metastasis after colon cancer surgery were analyzed by using univariate and multivariate logistic analysis. Kaplan-Meier risk curve was drawn, and log-rank test was used to analyze the predictive value of different HALP score for postoperative liver metastasis. Results HALP score were decreased in patients with maximum tumor diameter ≥5 cm, preoperative carcinoembryonic antigen (CEA) ≥5 μg/L, serous membrane and extrasserous infiltration, lymph node metastasis and vascular invasion, and the difference was statistically significant (P<0.05). Multivariate logistic regression analysis showed that HALP score [OR=1.467, 95%CI (1.253, 1.718), P<0.001], maximum tumor diameter [OR=3.476, 95%CI (1.475, 5.358), P=0.013], preoperative CEA level [OR= 6.197, 95%CI (2.436, 6.248), P=0.005], and lymph node metastasis [OR=2.593, 95%CI (1.667, 6.759) , P=0.003] were risk factors for postoperative liver metastasis of colon cancer. ROC curve analysis showed that the area under the curve of HALP score for predicting liver metastasis after colon cancer surgery was 0.908 (0.841, 0.974), the maximum value of the Youden index was 0.738, the optimal cut-off value of the HALP score was 35.5, the sensitivity was 0.852, the specificity was 0.886. Kaplan-Meier risk curve showed that the risk of early postoperative liver metastasis in the low HALP score group was higher than that in the high HALP score group (χ2=8.126, P=0.004). Conclusion Low HALP score in patients with colon cancer is associated with adverse prognosisi related pathological features, and is an influential factor for postoperative liver metastasis of colon cancer, and has predictive value for patients with postoperative liver metastasis 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.