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.
Objective To investigate effect of hepatocyte growth factor (HGF) after lentivirus-mediated RNA interference (RNAi) targeting c-Met on invasion of colonic carcinoma cell line SW480. Methods The experiment was assigned into 3 groups: NC group, the normal cells were infected by the shRNA negative control virus (the NC-20 andNC-40 represented the negative group which were added 20 ng/mL and 40 ng/mL respectively HGF after being infected); KD group, the normal cells were infected by the shRNA-c-Met target virus (the KD-20 and KD-40 represented the interfered group which were added 20 ng/mL and 40 ng/mL HGF respectively after being infected; KD1, KD2, KD3, and KD4 represented the different RNAi targets for the purpose gene); CON group, the normal cells were not infected by any virus. The lentiviral vector shRNA-c-Met was constructed and verified by polymerase chain reaction (PCR) and DNA sequencing. The SW480 cells were infected with the shRNA-c-Met after packed with lentivirus plasmid. Fourty-eight hours transfection later, the c-Met mRNA of the transfected SW480 cell was detected by real time PCR and the c-Met protein was examined by Western blot. Seventy-two hours after transfection, the cell apoptosis was detected by flow cytometry and the invasions in the different cells with stable transfection were detected by Transwell test. Results The RNAi sequence targeting c-Met gene was successfully inserted into the lentiviral vector. The shRNA-c-Met transfection resulted in an obviously reduced expression of c-Met mRNA in the SW480 cells. The efficency of gene knock down of the KD4 (the cells with No.4 target spot knocked down) was 81.4%. The shRNA-c-Met tansfection resulted in an obviously reduced expression of c-Met protein in the SW480 cells. After transfection, the apoptosis rate of the KD group was significantly higher than that in the NC group (P<0.001) or the CON group (P<0.001). The invasion ratios in the NC group, NC-20 group, and NC-40 group were significantly higher than those in the KD group (P<0.001), KD-20 group (P=0.015), and KD-40 group (P=0.017), respectively; which in the NC-20 group and NC-40 group were increased as compared with the NC group (P<0.001,P<0.001), and in the NC-40 group was increased as compared with the NC-20 group (P=0.005). The invasion ratios in the KD-20 group and KD-40 group were increased as compared with the KD group (P<0.001,P<0.001), and in the KD-40 group was increased as compared with the KD-20 group (P=0.014). Conclusion Lentivirus-mediated RNAi targeting c-Met could effectively suppress expression of c-Met in SW480 cells and could reduce invasion of HGF on SW480 cells with knocked down c-Met.
Objective To investigate the relationship between skin/pectoral muscle invasion and the prognosis of male breast cancer. Methods Clinical data and follow-up information of 79 male breast cancer patients who received treatment between September 2008 to April 2020 in West China Hospital were retrospectively reviewed, to analyze the clinicopathological features of male breast cancer and prognostic value of skin/pectoral muscle invasion. Results Among 79 male breast cancer patients, a total of 23 patients (29.1%) were with skin/pectoral muscle invasion at diagnosis. All the patients were followed up, with a median follow-up period of 63.3 months (1.0–204.5 months). Within follow-up period, 8 patients (10.1%) suffered from relapse, 19 patients (24.7%, 19/77) suffered from metastasis, and 4 patients (5.1%) died. Multivariate Cox proportional risk regression model suggested that patients with skin/pectoral muscle invaded had poor disease free survival [RR=4.48, 95%CI (1.08, 18.52), P=0.038]. Conclusions Skinor pectoral muscle invasion might be a valuable prognostic factor for male breast cancer patients. However, limited by sample size, the conclusion should be proved by further high-level studies.
ObjectiveTo establish a model for predicting microvascular invasion (MVI) of hepatocellular carcinoma based on magnetic resonance imaging (MRI) radiomics features.MethodsThe clinical and pathological datas of 190 patients with hepatocellular carcinoma who received surgical treatment in our hospital from September 2017 to May 2020 were prospectively collected. The patients were randomly divided into training group (n=158) and test group (n=32) with a ratio of 5∶1. Gadoxetate disodium (Gd-EOB-DTPA) -enhanced MR images of arterial phase and hepatobiliary phase were used to select radiomics features through the region of interest (ROI). The ROI included the tumor lesions and the area dilating to 2 cm from the margin of the tumor. Based on a machine learning algorithm logistic, a radiomics model for predicting MVI of hepatocellular carcinoma was established in the training group, and the model was evaluated in the test group.ResultsSeven radiomics features were obtained. The area under the receiver operating characteristic curve (AUC) of the training group and the test group were 0.830 [95%CI (0.669, 0.811)] and 0.734 [95%CI (0.600, 0.936)], respectively.ConclusionThe model based on MRI radiomics features seems to be a promising approach for predicting the microvascular invasion of hepatocellular carcinoma, which is of clinical significance for the management of hepatocellular carcinoma treatment.
Objective To explore the impact of microvascular invasion (MVI) on the survival prognosis of patients after radical hepatectomy for hepatocellular carcinoma, to analyze its related risk factors and preoperative prediction methods, and to provide reference and support for the treatment of early postoperative recurrence. MethodsBy searching domestic and international medical literature databases, we screened studies related to MVI in hepatocellular carcinoma, focusing on the definition, grading, risk factors, preoperative prediction methods, and postoperative treatment strategies of MVI, and summarized the results of the existing studies. ResultsMVI was a well-established risk factor for the intrahepatic metastasis and early postoperative recurrence of hepatocellular carcinoma. Currently, various methods were employed to predict MVI, including laboratory indicators, imaging genomics, and genomics. The laboratory indicators used for prediction included alpha-fetoprotein, protein induced by vitamin K absence or antagonist-Ⅱ, hepatitis B virus, tumor diameter, vascular endothelial growth factor A, and circulating tumor cells. Imaging genomics involved preoperative MRI with irregular tumor shape and intra-voxel incoherent motion diffusion-weighted imaging D value < 1.16 × 10-3 mm2/S, CT enhancement imaging features with irregular tumor margins, multiple foci, and contrast-enhanced ultrasound portal venous and delayed phase scores. Genomics included the maximum variant allele frequency of circulating tumor DNA. In cases where MVI was detected after surgery, adjuvant therapy options had gained attention, such as transcatheter arterial chemoembolization, hepatic arterial infusion chemotherapy, targeted therapy, immunotherapy, radiation therapy, antiviral therapy, and local treatment combined with systemic treatment. ConclusionsThe study of MVI and its targeted treatment strategies are important for reducing the postoperative recurrence rate of hepatocellular carcinoma and improving patient survival. The preoperative prediction model and postoperative treatment plan should be optimized in the future to provide more effective treatment reference for patients.
Objective To investigate the role of chemokine receptor CXCR7 in the development and progression of pancreatic carcinoma. Methods The short hairpin RNA (shRNA) targeting CXCR7 was designed and delivered into AsPC-1 pancreatic carcinoma cells to knock down CXCR7 expression. The cell proliferation, cell cycle, and apoptosis after CXCR7 knockdown was determined by MTT and flow cytometry, respectively. The invasive ability of pancreatic carcinoma cells was evaluated by using the Transwell system. Results Compared with the blank control group (BC group), transfection of AsPC-1 cells with CXCR7-shRNA resulted in a significantly decreased expression of CXCR7 at both mRNA and protein levels (P<0.05), and the ability of proliferation and invasion significantly decreased (P<0.05). Knockdown of CXCR7 also significantly increase apoptosis (P<0.05), induce cell cycle arrest at G0/G1 phase (P<0.05). Conclusions Taken together, the present study showes that the knockdown of CXCR7 expression may play an important role in pancreatic carcinoma development, invasion, and metastasis, CXCR7 may be a potential therapeutic target for the treatment of pancreatic carcinoma.
Objective To introduce the inflammatory microenvironment and epithelial-mesenchymal transition process of hepatocellular carcinoma, and review the relationship between them. Methods Domestic and international literatures were collected to summary the relationship between epithelial-mesenchymal transition and the inflammatory microenvironment of hepatocellular carcinoma. Result Many inflammatory factors and viral gene encoding proteins in the inflammatory microenvironment play an important role in the process of epithelial-mesenchymal transition in hepatocellular carcinoma. Conclusions The inflammatory microenvironment of hepatocellular carcinoma is an indispensable role in the process of epithelial-mesenchymal transition. The inhibition and treatment of inflammatory microenvironment may play a more active role in the control of tumor invasion and metastasis.
Objective To investigate the effect s of T lymphoma invasion and metastasis inducing factor 1 ( Tiam 1) antisense oligonucleotides (ASODN) on morphological remodeling of gast ric cancer cells. Methods The high-invasive and metastastic subgroup (MH ) was separated f rom human gast ric cancer cell line MKN245 (M0 ) by laminin adhesion method in vi t ro. And they were divided into four group s according to different further t reatment s : no t ransfection group (cont rol group ) , liposome t ransfection group , sense oligonucleotides2liposome t ransfection group ( SODN t ransfection with liposome group ) and antisense oligonucleotides2liposome t ransfection group (ASODN t ransfection with liposome group) . Then the expressions of Tiam 1 mRNA and protein were detected by RT-PCR and flowcytomet ry , respectively. The morphology changes between Tima 1 ASODN t ransfected MH cells and no t ransfected cells were observed by using HE stain , cytoskeletal protein stain and scanning elect ronic microscope (SEM) . Results Compared with the other group s , the expressions of Tiam 1 mRNA and protein in MH cells were significantly decreased af ter the cells were t ransfected with 0. 43 μmol/ L Tiam 1 ASODN ( P lt; 0. 01) . Additionally , it was observed that the t ransfected MH cells had less membrane surface projections , fewer or shortener pseudopodia , less irregular cytoskeletal network and less spotted-like actin bodys than no t ransfected MH cells did. Conclusion ASODN t ransfection could effectively suppress the expression of Tiam 1 and the remodeling in gast ric cancer cells , which may play an important role in the invasion and metastasis of gast ric cancer cells.
ObjectiveTo establish a model of chronic rotator cuff injury by inducing subacromial impingement syndrome in rats, so as to lay a foundation for further study on the mechanism and treatment of chronic rotator cuff injury. MethodsThe polyether-ether-ketone implants were designed and made with three-dimensional printing technique. In 48 male Sprague Dawley rats[weighing, (277.25±22.03) g], one shoulder joint was selected randomly as the experimental group; in the experimental group, the implant was pierced into shoulder joint close tothe acromion medial side and was pierced out under acromion close to the deltoid trailing edge; the penetratingpartof implant was cut off after the implanting part was fixed with 4# braided silk suture. No any treatment was performed on the contralateral shoulder of 24 rats as control group; and the implants were immediately removed after they were pierced into the contralateral shoulder joint of the other 24 rats as sham-operation group. All the rats were exposed to 30 minutes of downhill running at 17 m/minute (-13.5°) every day at 4 days after operation and the general condition of rats was observed. At 2, 4, 6, and 8 weeks after operation, 12 rats were sacrificed to collect shoulder joint samples. The hardtissueslices was used for VanGieson staining and histological observation of the supraspinatus tendon. ResultsAll ratssurvived to the end of experiment, without infection.The rats suffered limp at 2-3 daysafter operation, and the gait of most rats returned to normal at 4 days after operation.The histological results showed that the supraspinatus tendon had smooth edge, without split layers or breakage in the control group and sham-operation group. In the experimental group, the implants were positioned accurately without dislocation; 4 kinds of typical pathological changes were observed. Partial-thickness tear on bursal side appeared at 2 weeks (5 rats) and 4 weeks (2 rats), showing no significant difference between at 2 and 4 weeks (P > 0.05); intratendinous gap formed mainly at 4 weeks (10 rats) and 6 weeks (11 rats), showing significant differences when compared with that at 2 weeks (2 rats) (P < 0.05) and 8 weeks (2 rats) (P < 0.05); partial-thickness tear on articular side mainly appeared at 6 weeks (8 rats), showing significant difference when compared with that at other time points (P < 0.05); full-thickness tear was found mainly at 8 weeks (10 rats), showing significant differences when compared with that at other time points (P < 0.05). ConclusionA model of chronic rotator cuff injury is successfully established in rats through microinvasive implantation of subacromial impingement syndrome inducing implants and the pathological changes in this model are highly similar to the clinical pathological progress.
ObjectiveTo evaluate the prognostic significance of visceral pleural invasion in diameter 3-5 cm nonsmall cell lung cancer(NSCLC). MethodsA total of 112 patients who underwent lobectomy and pathologically diagnosed with NSCLC(3-5 cm) were included in our hospital between January 2006 and December 2010.There were 72 males and 40 females at average age of 61(28-72) years. There were 62 patients diagnosed as adenocarcinoma and 44 as squamous cell lung cancer. Viceral pleural invasion(VPI) was identified in 63 patients as a VPI group. The other 49 patients without VPI were as a NVPI group. All patients were performed with lobectomy and mediastinal lymph node dissection. ResultsThere was no perioperative mortality. More smokers were included in the VPI group when compared with the NVPI group(53.9% vs. 28.6%, P=0.007). More squamous cell cancers were included in the VPI group, while more adenocarcinoma were included in the NVPI group with a statistical difference(P=0.003). The average follow-up duration was 52 months. A total of 32 death occurred at the endpoint. The overall survival(OS) of all included patients was 71.4%. The average follow-up duration was 51 months in the VPI group and 54 months in the NVPI group(P=0.441). There was no statistical difference in OS between the VPI group and the NVPI group(61.7% vs. 83.7%, P=0.017). Cox regression showed age less than 65 years(P=0.007), TNM stage(P=0.013), and VPI(P=0.035) were significant prognostic factors for NSCLC. ConclusionWe identified the presence of VPI as an independent poor prognostic factor in NSCLC patients with diameter at 3-5 cm.