ObjectiveTo investigate the effects of pipecolic acid oxidase (PIPOX) on the proliferation, apoptosis, migration and invasion of primary liver cancer cells. MethodsImmunohistochemical staining and analysis of The Cancer Genome Atlas (TCGA) database were used to examine the PIPOX expression levels in liver cancer tissues and paired adjacent normal tissues, and studied their relationship with patient prognosis. Liver cancer cell lines stably overexpressing or knocking out PIPOX were constructed to explore PIPOX’s impact on liver cancer cell proliferation, apoptosis, migration and invasion by conducting in vitro functional experiments such as CCK-8, EdU, apoptosis detection, and Transwell assays. In vivo, nude mice subcutaneous tumor models and lung metastasis models were used to verify PIPOX’s effect on liver cancer growth and metastasis. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were both employed to detect the expression of epithelial-mesenchymal transition (EMT) markers in liver cancer cells. ResultsImmunohistochemical staining and TCGA database analysis revealed that PIPOX expression was significantly lower in liver cancer tissues compared to paired adjacent normal tissues (P<0.05). Prognostic analysis indicated shorter overall survival and disease-free survival in PIPOX low expression group (P<0.05). In vitro gain- and loss-of-function experiments showed that PIPOX significantly inhibited liver cancer cell migration and invasion (P<0.05), while having no significant effects on their proliferation and apoptosis (P>0.05). Animal experiments also confirmed that PIPOX significantly inhibited liver cancer lung metastasis (P<0.05), but had no significant effects on tumor growth (P>0.05). Finally, RT-qPCR and western blot results revealed that PIPOX promoted the expression of the epithelial marker E-cadherin (P<0.05) and inhibited the expression of mesenchymal markers (N-cadherin, vimentin, Snail) (P<0.05). ConclusionsPIPOX significantly inhibits liver cancer cell migration and invasion, potentially via suppressing the EMT process. However, PIPOX does not significantly affect liver cancer cell proliferation and apoptosis.
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.
ObjectiveTo investigate the influence of heat shock protein A2 (HSPA2) on the biological behavior of pancreatic adenocarcinoma cells and its mechanism. MethodsThe expressions of HSPA2 were determined in the human pancreatic adenocarcinoma cell lines (PANC-1, BxPC-3, and AsPC-1) using the Western blot. Subsequently, the cells with the lowest and highest HSPA2 expressions among these three lines were selected for conducting overexpression and knockdown experiments targeting HSPA2, respectively. The cellular proliferation, cell clonogenesis, migration, and invasion capabilities were assessed using MTT, clonogenic assay, and Transwell assay, respectively. Additionally, the impact of HSPA2 on the expression of key markers of epithelial-mesenchymal transition (EMT) was examined using the Western blot. The potential target molecules of HSPA2 were identified through immunoprecipitation assay and mass spectrometry. The rescue experiments further explored the regulatory relation between the HSPA2 and its target molecules. The influence of HSPA2 on pancreatic adenocarcinoma growth was investigated through establishment of xenograft tumor model in nude mice. ResultsThe HSPA2 exhibited the lowest expression in the PANC-1 cells and the highest expression in the AsPC-1 cells among the three cell lines. Subsequent functional studies demonstrated that the overexpression of HSPA2 in the PANC-1 cells markedly promoted proliferation, cell clonogenesis, migration, and invasion, while the knockdown of HSPA2 expression in the AsPC-1 cells markedly inhibited these processes. The Western blot analysis further showed that the HSPA2 overexpression downregulated E-cadherin expression and upregulated N-cadherin and Vimentin expressiones, whereas the HSPA2 knockdown produced opposite effects. The rescue experiments indicated that the HSPA2 promoted the EMT in pancreatic adenocarcinoma cells by upregulating Yes associated protein (YAP). The subcutaneous xenograft tumor experiments in the nude mice showed that the HSPA2 knockdown inhibited tumor growth. ConclusionThe results of this study suggest that HSPA2 promotes EMT via upregulating YAP, which facilitates proliferation, migration, and invasion of pancreatic adenocarcinoma cells.
ObjectiveTo investigate expressions and biological function of membrane type matrix metallopro-teinase-1 (MT1-MMP) and matrix metalloproteinase-2 (MMP-2) in papillary thyroid carcinoma. MethodsThe expre-ssions of MT1-MMP and MMP-2 in 164 cases of papillary thyroid carcinoma and paracancerous tissues were detected by immunohistochemistry.The association between the expressions of MT1-MMP and MMP-2 and clinicopathological characteristics of papillary thyroid carcinoma was analyzed. ResultsIn paracancerous tissues, the positive expression rate of MT1-MMP was 11.0% (18/164), and the positive expression rate of MMP-2 was 14.0% (23/164).In papillary thyroid carcinoma tissues, the positive expression rates of MT1-MMP and MMP-2 was 61.6% (101/164) and 67.7% (111/164), respectively.The expressions of MT1-MMP and MMP-2 in carcinoma tissues and para carcinoma tissues were statistically significant differences (P < 0.05).The expressions of MT1-MMP and MMP-2 in papillary thyroid carcinoma tissues correlated with the lymph node metastasis (P < 0.05).In addition, the expression of MMP-2 in papillary thyroid carcinoma tissues correlated with capsule invasion (P < 0.05).The positive correlation was found between the expressions of MT1-MMP and MMP-2 in papillary thyroid carcinoma tissues (r=0.256, P < 0.05). ConclusionsMT1-MMP and MMP-2 may be involved the thyroid capsule invasion and lymph node metastasis of papillary thyroid carcinoma.MT1-MMP and MMP-2 may be involved in the progression of papillary thyroid carcinoma.
ObjectiveTo assess the efficiency, safety and long-term prognosis for interventional bronchoscopy in the treatment of trachea invasion by thyroid cancer. MethodsThe clinical data of forty-three patients with trachea invasion by thyroid cancer in Changhai Hospital from January 2006 to September 2015 were retrospectively analyzed. The trachea diameter and dyspnea score were compared before and after interventional treatment to explore the efficiency. The complications during and after therapy were observed. All patients were treated with interventional modalities including electrocautery, argon plasma coagulation, laser, cryotherapy, stent insertion or radioactive seeds implantation according to different invasion types, degree of stenosis and base situation. ResultsThe trachea diameter increased from (3.9±1.5)mm to (10.6±0.6)mm after bronchoscopy therapy (t=-17.314, P < 0.000 1). The dyspnea score decreased from 3.3±0.7 to 2.3±0.7 after bronchoscopy therapy (t=9.274, P < 0.000 1). The complications during therapy included haemorrhage (46.5%), vocal cord paralysis (4.7%) and glottis edema (7.0%). The restenosis rate in the patients with stent insertion was 26.7%. Thirty-seven patients were followed up successfully, and the medium survival time for follow-up patients was 27 months. The univariate and multivariate analysis indicated that the kind of interventional modalities used for therapy was an independent prognostic factor of survival (HR=0.261, P=0.036). The medium survival time for the patients treated with≥3 methods, 2 methods and 1 method was 47 months, 36 months and 13 months, respectively. ConclusionsFor trachea invasion by thyroid cancer, bronchoscopic therapy can effectively relieve airway obstruction and dyspnea symptom. Combination of multiple interventional modalities could have a favorable prognosis after treatment.
Objective To expand the utilization of minimally invasive technologies for parapancreatic abscess, and summarize the application experience of choledochoscope for treatment of parapancreatic abscess. Methods The clinical data and treatment effectiveness of 36 patients with parapancreatic abscess from Dec. 2000 to Dec. 2008 were analyzed retrospectively. These patients had experienced percutaneous puncture and been placed drainage tube under the ultrasound guidance first, then expanded the sinus tract gradually, and performed debridement by choledochoscope. The flexibility of choledochoscope was used to remove the necrotic tissue and pyogenic membrane repeatedly by clamping, netting and vacuum aspiration in every domain. Results Thirty-six patients were performed percutaneous puncture and placed drainage tube, 3 cases were given canalis singularis, 7 cases were double tube, 26 cases were over three tube. The debridement times were 3-14 by choledochoscope, average 5.6 times. There were 6 cases with improving systemic symptoms, blood routine and temperature recovering normal, and drink and food recovering, then discharged from hospital with tube after 1-2 times of debridement. Length of stay was 25-132 d, average 76 d. The curing rate was 91.7% (33/36). Two cases were turned into open surgery because of broad necrotic tissue range combined with many abdominal cavity abscess with good postoperative recovery and cured. One case was dead of severe multiple organ failure combination. There were 2 patients with hemorrhage, 3 patients with external intestinal fistula. Conclusions The debridement of choledochoscope for parapancreatic abscess treatment is a simple, flexible and effective method. It changes the viewpoint that parapancreatic abscess can be cured only by operation drainage, decreases the patients’ trauma and accomplishes the idea of damage control by minimally invasive technologies.
Objective To evaluate the clinical value of ureteroscope in cholelithiasis treated by laparoscopic surgery. Methods The clinical data of 36 patients admitted because of hepatolithus with ureteroscope combination in laparoscopic surgery from February 2007 to September 2009 in Guidong People’s Hospital of Guangxi were analyzed retrospectively. Results In 33 cases, stones were removed once by ureteroscope in laparoscopic surgery with residual stones (in 3 cases residual stone were removed secondarily through T tube) and the other 3 cases were transferred to laparotomy forcedly due to bleeding of biliary duct and vessels of porta hepatis and tearing of bile duct. During operation, blood loss was 30-280 (94.51±54.70) ml; operation time was 110-260 (147.22±48.45) min; recovery time of bowel movement was 1-3 (2.03±0.76) d; postoperative hospitalization time was 6-13 (7.12±1.65) d (some discharged with T tube); the time of patients of T tubes pulled out was 28-45 (38.92±6.52) d. Bile leakage happened in 1 case and infection of biliary tract in 1 case, no complications such as biliary stricture or bile duct bleeding were found after operation. Conclusions Treatment of intrahepatic bile duct or a single extra-hepatic sand-like stones with ureteroscopy usage in laparoscopic surgery is feasible and less invasive. It is a minimally invasive treatment for intra- or extra-hepatic stones due to rapidly postoperative rehabilitation.
ObjectiveTo investigate the regulatory mechanism of thioredoxin binding protein (TXNIP)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) pathway in the occurrence and development of breast cancer.MethodsThe resected 15 cases of breast cancer tissues and their adjacent tissues in our hospital from September 2019 to June 2020 were selected, and the immunohistochemistry was used to detect the expression levels of TXNIP and NLRP3 in breast cancer and its adjacent tissues. Three kinds of breast cancer cell lines (MDA-MB231, MCF-7 and SKBR3) and normal breast epithelial cell line (HMEC) were collected. Western blot was used to detect the relative expression levels of TXNIP and NLRP3 in three kinds of breast cancer cell lines and HMEC cell line. MDA-MB231 cancer cells were divided into blank control group (normal culture without any treatment), TXNIP overexpression group (Ad-TXNIP group, transfected with adenovirus vector carrying TXNIP overexpression sequence), Ad-TXNIP negative control group (Ad-eGFP1 group, transfected of empty adenovirus vector without TXNIP overexpression sequence), NLRP3 overexpression group (Ad-NLRP3 group, transfected with adenovirus vector containing NLRP3 overexpression sequence), TXNIP and NLRP3 overexpression co-transfection group (Ad-TXNIP+Ad-NLRP3 group, co-transfection of adenovirus vector carrying TXNIP and NLRP3 overexpression sequence), TXNIP overexpression and Ad-NLRP3 negative control (Ad-eGFP2) co-transfection group (Ad-TXNIP+Ad-eGFP2 group,co-transfection of adenovirus vector carrying TXNIP overexpression sequence and empty adenovirus without NLRP3 overexpression sequence). After 24 hours of transfection and culture, CCK-8 method was used to detect the MDA-MB231 cells proliferation. Transwell chamber method was used to detect MDA-MB231 cells migration and invasion. Nude mice tumorigenicity test was used to detect the tumorigenicity of the MDA-MB231 cells in vivo. Western blot was used to detect the expressions of TXNIP, NLRP3, proliferation marker protein (Ki-67), caspase-1, vascular endothelial growth factor (VEGF), interleukin (IL)-1β, IL-18 and caspase-1 precursor protein (pro-caspase-1) in the MDA-MB231 cells.ResultsCompared with the adjacent tissues, the relative expression level of TXNIP decreased (P<0.05) and the relative expression level of NLRP3 increased (P<0.05) in breast cancer tissues. Compared with normal breast epithelial cell line (HMEC cell line), the relative expression levels of TXNIP in MDA-MB231, MCF-7 and SKBR3 breast cancer cell lines were decreased (P<0.05), and the relative expression levels of NLRP3 were increased (P<0.05). Compared with the blank control group, the relative expression levels of TXNIP, NLRP3, IL-1β, IL-18, pro-caspase-1 and caspase-1 were increased (P<0.05), the relative expression levels of Ki-67 and VEGF, the proliferation activity, invasion and migration ability of MDA-MB231 cells and tumor weight were decreased (P<0.05) in the Ad-TXNIP group and the Ad-NLRP3 group. Compared with the Ad-TXNIP group and the Ad-NLRP3 group, the relative expression levels of TXNIP, NLRP3, IL-1β, IL-18, pro-caspase-1 and caspase-1 were further increased (P<0.05), the relative expression levels of Ki-67 and VEGF, the proliferation activity, invasion and migration ability of MDA-MB231 cells and tumor weight were further decreased (P<0.05) in the Ad-TXNIP+Ad-NLRP3 group.ConclusionsIn breast cancer tissues and breast cancer cell lines, TXNIP is low expression and NLRP3 is high expression. They can interact with each other to promote pyroptosis and inhibit the proliferation, invasion and migration of breast cancer cells.
ObjectiveTo investigate relationship between galectin-1 (Gal-1) and invasion and metastasis of gastric cancer.MethodThe related literatures on the relationship between the Gal-1 and the invasion and metastasis of gastric cancer in recent years were reviewed.ResultsThe expression of Gal-1 in the gastric cancer tissue was closely related to the invasion, metastasis ability, immunosuppression, and angiogenesis of tumor cells, and played the important role in the progression and evolution of the gastric cancer. The expression of Gal-1 was higher, the malignancy of the tumor was higher, and the prognosis of the patient was worse.ConclusionExpression of Gal-1 can promote invasion and metastasis of gastric cancer, but further clinical trials need to be verified.
ObjectiveTo study the mechanism of the effect on invasion and metastasis of colorectal cancer by down-regulating c-Met gene.Methodsc-Met genes were knocked down in SW480 cells, differential genes were screened by gene chip, functional cluster analysis of differential genes was carried out, and IPA was used to analyze the interaction network of cell signal pathway and related differential genes, as well as the ralationship between related genes and upstream regulatory molecules. The related genes in the suppressed signal pathway were selected for qPCR verification.ResultsAfter knockdown of c-Met, the number of up-regulated genes and down-regulated genes in SW480 cells was 399 and 286, respectively. Cluster analysis showed that c-Met knockdown had a great effect on the gene expression level of SW480 cells, IPA pathway analysis showed that HGF signaling pathway was suppressed, and after c-Met knockdown, IPA interaction network suggested that AKT2, PIK3CA, and MAP2K4 in HGF pathway were down-regulated, and qPCR verified that the above genes were also down-regulated, which was consistent with the results of microarray.Conclusionc-Met may affect the invasion and metastasis of colorectal cancer through the regulation of AKT2, PIK3CA, and MAP2K4 in HGF pathway.