摘要
Introducción. La enfermedad hepática esteatósica asociada a disfunción metabólica (MASLD) se ha convertido en la enfermedad hepática crónica más frecuente en los países occidentales, causando un aumento en los costos y en la ocupación hospitalaria. La caracterización integral previa al trasplante hepático en pacientes con MASLD es una gran interrogante, especialmente en nuestro medio. El objetivo del presente estudio fue realizar la caracterización clínico-epidemiológica de pacientes trasplantados por cirrosis hepática (CH) descompensada o carcinoma hepatocelular (CHC) asociado a MASLD. Metodología. Se desarrolló un estudio observacional retrospectivo, descriptivo, de corte transversal en el Servicio de Hepatología del Hospital Pablo Tobón Uribe en Medellín, Colombia. Se incluyeron pacientes mayores de 17 años, con diagnóstico de CH o de CHC asociado a MASLD que fueron trasplantados entre los años 2004 a 2017. Resultados. Se encontraron 84 pacientes que fueron trasplantados con esas características. La edad promedio de los pacientes fue de 59±10,5 con una mayor proporción significativa de hombres sobre mujeres, llegando casi al 70 %. Con relación a las comorbilidades, se encontró que el sobrepeso/obesidad, la hipertensión arterial y la diabetes mellitus tipo 2 fueron un hallazgo en el 44,1 %, 33,3 % y 33,3 %, respectivamente. Por otro lado, el 14,5 %, el 33,7 % y el 51,8 % presentaron un Child-Pugh A, B y C, respectivamente. La media del puntaje MELD fue de 18,9±6,26. Con respecto a las complicaciones de la cirrosis, el 77,4 % de los pacientes presentó ascitis, el 61,9 % encefalopatía hepática, el 36,9 % hemorragia del tracto digestivo superior y el 29,8 % peritonitis bacteriana espontánea. Conclusión. Los resultados expuestos mostraron nuestra experiencia en trasplante hepático en pacientes con CH y CHC asociado a MASLD. Se debe realizar una evaluación multidisciplinaria antes y después del trasplante en estos pacientes, haciendo especial énfasis en el manejo de la disfunción metabólica y sus componentes, entre los que se destacan la obesidad y la diabetes mellitus.
Introduction. Metabolic dysfunction-associated steatotic liver disease (MASLD) has become the most frequent chronic liver disease in Western countries, causing increased costs and hospital occupancy. The comprehensive pre-transplant characterization in patients with MASLD is a major question, especially in our setting. The aim of the present study was to perform the clinical-epidemiological characterization of transplanted patients with decompensated liver cirrhosis (LC) or hepatocellular carcinoma (HCC) associated with MASLD. Methodology. A retrospective, descriptive, cross-sectional observational study was carried out in the Hepatology Department of the Pablo Tobón Uribe Hospital in Medellin, Colombia. Patients over 17 years of age, with a diagnosis of LC or HCC associated with MASLD who were transplanted between 2004 and 2017 were included. Results. We found 84 patients who were transplanted with these characteristics. The mean age of the patients was 59±10.5 with a significantly higher proportion of men over women, reaching almost 70%. Regarding comorbidities, overweight/obesity, arterial hypertension, and type 2 diabetes mellitus were found in 44.1%, 33.3%, and 33.3%, respectively. On the other hand, 14.5%, 33.7%, and 51.8% had Child-Pugh A, B, and C, respectively. The mean MELD score was 18.9±6.26. Regarding complications of cirrhosis, 77.4% of patients developed ascites, 61.9% hepatic encephalopathy, 36.9% upper gastrointestinal tract hemorrhage, and 29.8% spontaneous bacterial peritonitis. Conclusion. The above results showed our experience of liver transplantation in patients with LC and HCC associated with MASLD. A multidisciplinary evaluation should be performed before and after transplantation in these patients, with special emphasis on the management of metabolic dysfunction and its components, including obesity and diabetes mellitus.
Subject(s)
Non-alcoholic Fatty Liver Disease摘要
Small nucleolar RNAs (snoRNAs) are non-coding RNAs in the nucleolus and are mainly responsible for the 2'-O-methylation and pseudouridine modification of rRNA. snoRNAs regulate various biological processes, such as tRNA modification, spliceosome snRNA modification, maintenance of the telomere structure, and alternative splicing of mRNA. Aberrant expression of snoRNA is related to cancer progression, and it may become a new target for cancer treatment. snoRNAs are stable and easy to detect in body fluids, so they can be used as a biomarker for clinical diagnosis and prognostic. This article reviews the biogenesis, classification, structure, and function of snoRNAs and introduces their potential role in the occurrence and development of hepatocellular carcinoma.
摘要
Liver transplantation is the optimal treatment for end-stage liver disease and hepatocellular carcinoma, which can significantly improve clinical prognosis and quality of life of patients. However, multiple challenges, such as rejection, immune tolerance, shortage of donor liver, preservation of donor liver, ischemia-reperfusion injury and postoperative complications, <i>etc.</i>, limit the efficacy of liver transplantation in clinical practice. Research teams in China have made significant contributions to the basic research related to liver transplantation by making continuous efforts and combining the development of emerging technologies, interdisciplinary integration and other emerging fields. In this article, the frontier progress in the basic research of liver transplantation in 2023 was reviewed, highlighting the progress made by Chinese research teams in the basic research of liver transplantation, aiming to provide reference for promoting the integration of Chinese characteristics into the research of liver transplantation, accelerate the integration of Chinese liver transplantation research with international community, and promote further advancement of liver transplantation in China.
摘要
As a mature organ transplantation, liver transplantation has become the optimal treatment for end-stage liver disease, which can improve the quality of life of recipients. However, liver transplantation still faces multiple challenges, such as rejection, infection, biliary complications, delayed graft function, ischemia-reperfusion injury, recurrence of hepatocellular carcinoma after liver transplantation, kidney-related diseases after transplantation, and donor shortage, <i>etc.</i>, which remain to be improved and urgently resolved. With persistent attempts and experience accumulated by Chinese experts and scholars, these problems related to liver transplantation have been gradually resolved year by year. In 2023, liver transplantation teams in China have achieved a series of significant progresses in the field of clinical research. In this article, clinical frontiers and novel technological progresses in the field of liver transplantation in 2023 were reviewed, and the achievements of clinical liver transplantation in China in 2023 were summarized, aiming to provide novel ideas for promoting further development of liver transplantation in China.
摘要
ObjectiveTo explore the effects of modified Danggui Beimu Kushen pills on tumor growth and T-cell subsets in H22 hepatocellular carcinoma-bearing mice and to provide an experimental basis for the treatment of hepatocellular carcinoma with modified Danggui Beimu Kushen pills combined with immune checkpoint antibodies. MethodA H22 hepatocellular carcinoma-bearing mouse model was established. The modeled mice were randomized into model, cisplatin, low- (4 g·kg-1·d-1), medium- (8 g·kg-1·d-1), and high-dose (16 g·kg-1·d-1) modified Danggui Beimu Kushen pills groups. After continuous administration for 14 days, the mice were sacrificed on day 15. The tumor volume was measured on days 0, 4, 8, 12, 15 of drug administration. Tumors were weighed and thymus index and spleen index were calculated. Spleen lymphocytes were co-cultured with H22 hepatoma cells, and the tumor cell-killing rate was detected by the cell counting kit-8 (CCK-8). Real-time polymerase chain reaction was carried to determine the mRNA levels of programmed cell death protein-1 (PD-1) and lymphocyte activation gene-3 (LAG-3) in spleen and tumor tissues. The number of CD4+ and CD8+ T cells and the expression of PD-1 and LAG-3 were detected by immunohistochemistry (IHC). ResultOn day 8 of drug administration, tumor volumes in all treatment groups decreased compared with that in the model group. On day 15, both tumor volume and tumor weight were significantly lower in the treatment groups than in the model group (P<0.01), with the cisplatin group showing the most pronounced reduction. Compared with the model and cisplatin groups, medium- and high-dose modified Danggui Beimu Kushen pills increased the thymus index (P<0.01). Compared with the model group, all treatment groups showed increased spleen index (P<0.05, P<0.01), with the cisplatin group showing the most significant increase. Compared with the model and cisplatin groups, all the groups of modified Danggui Beimu Kushen pills demonstrated increased number of CD4+ and CD8+ T cells and tumor cell-killing rate in the spleen and tumor tissues (P<0.01) and down-regulated mRNA and protein levels of LAG-3 (P<0.05, P<0.01). The high-dose group of modified Danggui Beimu Kushen pills had lower mRNA level of PD-1 in the tumor tissue than the model and cisplatin groups (P<0.01). ConclusionModified Danggui Beimu Kushen pills may promote the proliferation and tumor microenvironment infiltration of CD4+ and CD8+ T cells in H22 tumor-bearing mice by down-regulating LAG-3 expression, thereby improving T-cell immune activity and inhibiting tumor growth. This study provides an experimental basis for the combination of modified Danggui Beimu Kushen pills and immune checkpoint antibodies in the treatment of hepatocellular carcinoma.
摘要
Objective:To screen the interacting protein of ubiquitin-conjugating enzyme E2S(UBE2S)and construct the hepatocellular carcinoma(HCC)based on UBE2S interacting protein prognosis model(UIPM),and to discuss the value of UIPM in assessing the prognosis of the HCC patients.Methods:Co-immunoprecipitation(Co-IP)was used to screen the protein complexes binding to Flag-UBE2S.After validation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis(SDS-PAGE)and Western blotting methods;liquid chromatography-mass spectrometer(LC-MS)was used to identify the UBE2S interacting proteins;Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were conducted on these proteins;the prognosis-related proteins from The Cancer Genome Atlas(TCGA)were cross-referenced with UBE2S interacting proteins by survival package of R software;the key proteins were extracted through LASSO regression analysis to build the UIPM;the prognostic model risk scoring formula was established.The HCC patients in TCGA were divided into high risk group and low risk group based on median value of the risk scores.The predictive accuracy of UIPM was evaluated by receiver operating characteristic curve(ROC),and the predictive accuracy was further validated by International Cancer Genome Consortium(ICGC)Database;univariate regression analysis and multivariate Cox regression analysis were used to detect whether the UIPM risk score was an independent prognostic factor for HCC.Furthermore,the nomogram model was built.Results:A total of 97 UBE2S interacting proteins were identified through Co-IP combined with LC-MS analysis.The GO functional enrichment analysis and KEGG signaling pathway enrichment analysis results showed that the interacting proteins were closely associated with cysteine-type endopeptidase activity,oxidative stress,and cell death.The TCGA revealed 5 163 HCC prognosis-related proteins;after intersecting with UBE2S interacting proteins,40 prognosis-related interacting proteins were found.Seven key proteins were determined through LASSO regression analysis,including UBE2S,heat shock protein family A member 8(HSPA8),heterogeneous nuclear ribonucleoprotein H1(HNRNPH1),chaperonin containing TCP1 subunit 3(CCT3),eukaryotic translation initiation factor 2 subunit 1(EIF2S1),receptor for activated C kinase 1(RACK1),and actin related protein 2/3 complex subunit 4(ARPC4),and the UIPM was constructed.There was significant difference in survival rate of the patients between high risk group and low risk group(P<0.05).The ROC curve analysis results showed the area under ROC curve(AUC)values of UIPM for predicting 1-year,2-year,and 3-year survival risk scores of the HCC patients were all greater than 0.7,indicating the model had high predictive accuracy.This was also confirmed by ICGC Database data.The univariate and multivariate Cox regression analysis results showed that the UIPM risk score was an independent prognostic risk factor for the HCC patients(P<0.05).The nomogram results showed good consistency between predicted survival rate and actual survival rate of the patient.Conclusion:A total of 97 interacting proteins that interact with UBE2S may promote the occurence and devolopment of HCC through oxidative stress and dysregulation of ferroptosis pathways.The UIPM risk score is an independent risk factor for the prognosis of HCC and can be used to predict the outcomes of the patients.UBE2S,HSPA8,HNRNPH1,CCT3,EIF2S1,RACK1,and ARPC4 could be regarded as the new biomarkers and therapeutic targets for HCC.
摘要
Objective:To discuss the factors related to the prognosis in the alpha fetoprotein(AFP)negative hepatocellular carcinoma(HCC)patients,and to construct the nomogram for predicting the survival time of the patients.Methods:The retrospective analysis on data of 2 064 cases of AFP negative HCC patients extracted from the Surveillance,Epidemiology,and End Results(SEER)Database was conducted,and all the patients were divided into training cohort and internal validation cohort at a ratio of 7∶3,and 101 AFP negative HCC patients from the Integrated Traditional Chinese and Western Medicine Hospital in Hunan Province were regarded as the external validation cohort.The univariate Cox regression analysis results were incorporated into the multivariate analysis,and the independent risk factors for the AFP negative HCC patients were obtained by multivariate Cox analysis to build a cancer specific survival(CSS)prognosis nomogram for the AFP negative HCC patients.The predictive efficacy and clinical utility of the nomogram were evaluated by time-dependent receiver operating characteristic curve(ROC),calibration plots,and decision curve analysis(DCA).The total score obtained from the nomogram was used for the risk stratification to compare the degree of risk discrimination between the nomogram and the American Joint Committee on Cancer(AJCC)staging system.Results:Ten independent risk factors were selected by multivariate Cox regression analysis to construct 3-year,4-year,and 5-year CSS prognostic nomograms for the AFP negative HCC patients,including the patient's age,pathological grade,surgical status,radiotherapy status,chemotherapy status,lung metastasis status,tumor size,tumor T stage,tumor M stage,and marital status.The area under curve(AUC)for the 3-year,4-year,and 5-year time-dependent ROC in the training cohort were 0.807(95%CI:0.786-0.828),0.804(95%CI:0.782-0.826),and 0.813(95%CI:0.790-0.835),respectively.In the internal validation cohort,they were 0.776(95%CI:0.743-0.810),0.772(95%CI:0.737-0.808),and 0.789(95%CI:0.752-0.826),and in the external validation cohort,they were 0.773(95%CI:0.677-0.868),0.746(95%CI:0.620-0.872),and 0.736(95%CI:0.577-0.895).The calibration plots verified that the nomogram fitted well with the perfect line.The DCA curve revealed that the net benefit of the nomogram was significatly higer than that of the AJCC staging system at certain probability thresholds compared with AJCC staging,the nomogram had a better ability to identify high-risk individuals.Conclusion:The serum AFP expression is one of the prognostic markers for the HCC patients.For those patients with AFP negative expression in serum,different considerations should be taken.The nomogram model based on multiple risk factors is a promising clinical tool for assessing the CSS in the AFP negative HCC patients.
摘要
ObjectiveTo investigate the effect of Shugan Quyu Jiedu prescription (SGQYJDF) on inducing ferroptosis in hepatocellular carcinoma cells based on the tumor protein 53 (p53)/solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) pathway. MethodMHCC97H cells were divided into the blank serum group (10% blank serum medium), SGQYJDF-containing serum low concentration group (5% SGQYJDF-containing serum and 5% blank serum medium), SGQYJDF-containing serum medium concentration group (7.5% SGQYJDF-containing serum and 2.5% blank serum medium), SGQYJDF-containing serum high concentration group (10% SGQYJDF-containing serum medium) and sorafenib group (sorafenib concentration of 10 μmol·L-1 in 10% blank serum medium). After 24 hours of intervention, the cell survival rate was detected by cell counting kit-8 (CCK-8) assay. The cell proliferation ability was detected by 5-ethynyl-2′-deoxyuridine (EdU) staining. The intracellular ferrous ion (Fe2+) level was detected by ferrous ion fluorescent probe (FerroOrange) staining. The intracellular malondialdehyde (MDA) and glutathione (GSH) levels were detected by colorimetric assays. The ultrastructure of mitochondria was observed by transmission electron microscopy. The expression levels of ferroptosis-related proteins p53, SLC7A11 and GPX4 were detected by Western blot. ResultIn terms of cell viability, compared with the blank serum group, the SGQYJDF group showed a dose-dependent decrease in the survival rate of MHCC97H cells. Effect of the medium and high concentrations of SGQYJDF on the survival rate of MHCC97H cells were significantly decreased (P<0.01). Additionally, the results of the EdU assay showed that both the medium and high concentrations of SGQYJDF were able to inhibit the proliferation ability of MHCC97H cells (P<0.05, P<0.01). Regarding the biochemical indicators of ferroptosis, compared to the blank serum group, the medium and high concentrations of SGQYJDF were able to dose-dependently increase the intracellular Fe2+ level (P<0.01). The low, medium, and high concentrations of SGQYJDF were able to dose-dependently decrease the level of GSH in MHCC97H cells (P<0.01) and increase the level of MDA in the cells (P<0.05, P<0.01). In terms of pathway-related protein expression, compared to the blank serum group, the medium and high concentrations of SGQYJDF could significantly increase the expression of p53 (P<0.01). The low, medium, and high concentrations of SGQYJDF could significantly decrease the expression of GPX4 (P<0.01). The high concentration of SGQYJDF could decrease the expression of SLC7A11 (P<0.01). In terms of the cell morphology of ferroptosis, compared with the blank serum group, transmission electron microscopy revealed that the low concentration of SGQYJDF caused mitochondrial deformation, while the medium and high concentrations of SGQYJDF resulted in reduced mitochondrial volume, increased double-layer membrane density, and decreased mitochondrial cristae. These features were similar to those of sorafenib-induced ferroptosis. Furthermore, compared with the sorafenib group, the high concentration of SGQYJDF showed no statistically significant differences in cell survival rate, proliferation ability, Fe2+ level, MDA level, and GSH level. ConclusionThe results suggest that SGQYJDF may induce ferroptosis and inhibit proliferation in hepatocellular carcinoma MHCC97H cells by upregulating the expression of p53, suppressing the expressions of GPX4 and SLC7A11, downregulating the level of GSH, and leading to the accumulation of intracellular Fe2+ and MDA.
摘要
Aim To explore the new mechanism of triptolide (TRI) inhibiting the progression of hepatocellular carcinoma (HCC) . Methods Different concentrations (0, 0 . 5, 2, and 8 jjunol • L~) of TRI were administered to act on liver cancer cells, and then the cell phenotypes and possible mechanisms were explored using experimental methods such as CCK-8, cell cloning, Transwell, and protein immunoblotting; siRNA was used to interfere with the target gene GSDME and its role was determined. Finally, the mechanism of TRI inhibiting the growth of HCC cells in vivo was validated using a transplanted tumor model. Results TRI could inhibit the proliferation, cloning, and invasion of HCC cells, and promote cell apoptosis. Immunoblotting results showed that the expression of GSDME was significantly upregulated in HepG2 or He-pal-6 hepatocellular carcinoma after TRI treatment, while the expression of cleaved caspase-3 and PARP also significantly increased. Knocking out GSDME could partially reverse TRI-induced cell apoptosis. At the same time, cells knocked down by GSDME had stronger cloning and migration abilities, and the apoptosis rate was reduced compared to the TRI treatment group alone. In vivo experiments showed that TRI inhibited HCC tumor growth, and the TRI + siGSDME group had a faster tumor growth rate than the TRI treatment group alone did. In addition, after TRI stimulation, p-eIF2a and ATF4 in HepG2 and Hepal-6 cells significantly increased. The immunofluorescence results showed a dose-dependent increase in the number of ATF4 positive cells in HepG2 and Hepal-6 cells after TRI stimulation. Conclusion The inhibitory effect of TRI on the growth and invasion of liver cancer cells may be related to its regulation of the ATF4/caspase-3/GSDME signaling pathway and promotion of liver cancer cell apoptosis.
摘要
ObjectiveSorafenib is a first-line only drug approved for the treatment of advanced hepatocellular carcinoma (HCC). Resistance to sorafenib means that treatment outcomes are often unsatisfactory. Although the mechanism underlying sorafenib resistance remains unclear, resistance may occur through Akt signaling pathway activation in HCC. Dihydrotanshinone (DHT), a lipophilic component of traditional Chinese medicine Salvia miltiorrhiza Bunge, has multiple anti-tumor activities and inhibits Akt activation. The effect and mechanism of DHT combined with sorafenib on HCC have not been investigated. In this study, we investigate whether DHT potentiates the anti-cancer activities of sorafenib against HCC. MethodsIn this study, the effects of sorafenib and DHT on the viability, apoptosis and drug sensitivity of Huh7 and HepG2 cells were verified by Cell Counting Kit-8 (CCK-8) and flow cytometry. Akt, P-Akt, Caspase3, GSK-3β, P-GSK-3β, ribosomal protein S6 kinase (S6K), P-S6K, Cyclin D1, Bcl-xl, Bcl-2, and Bax expression levels were analyzed via Western blot. All data were statistically compared using one-way analysis of variance (ANOVA) and Dunnett test. Statistical analysis using SPSS 20.0 statistical software. ResultsDHT inhibit proliferation and promote apoptosis in HCC cells by reducing Akt activation. DHT inhibits the expression and activation of Akt downstream factors, including GSK-3β and S6K, which regulate the apoptotic response and are activated and upregulated by sorafenib treatment. Both sorafenib and DHT downregulate cyclin D1 expression and DHT upregulates Bax expression and downregulates Bcl-2 and Bcl-xl expression. However, sorafenib had little influence on Bcl-2 family protein expression. ConclusionDHT may enhance the proliferation inhibition and apoptosis induction of sorafenib in HCC cells by inhibiting the activation of Akt signaling pathway, thus enhancing the anticancer effect of sorafenib.
摘要
Hepatocellular carcinoma (HCC) is one of the cancers with the highest incidence and mortality rates in China and often presents with insidious early clinical manifestations. This frequency results in the majority of patients being diagnosed at middle and advanced stage of the disease, thereby missing the opportunity for potentially curative surgical interventions. For patients who are ineligible for radical surgical resection, a variety of therapeutic approaches, including systemic antitumor therapy, local radiotherapy, interventional treatment, and liver transplantation, have been employed. Moreover, neoadjuvant therapies have transformed a subset of initially unresectable HCC cases into operable ones. Nevertheless, many patients fail to benefit from these treatments, underscoring the urgent need for novel therapeutic targets. Cancer-associated fibroblasts (CAFs), a principal component of the solid tumor microenvironment, play a pivotal role in the proliferation, migration, invasion, and treatment resistance of cancer cells. This review delineates the origins of CAFs and their mechanisms of action in the pathogenesis and progression of HCC and discusses potential therapeutic strategies targeting CAFs.
摘要
Objective @#To study the molecular mechanism of fat mass and obesity⁃associated protein (FTO) regulating hepatocellular carcinoma (HCC) . @*Methods@#HepG2 cells of knock⁃down FTO were constructed , HepG2 cells of knock⁃down FTO and HepG2 cells were collected , and high⁃throughput sequencing was performed using Illumina Hiseq platform to screen the gene expression differences between the two groups . Through GO and KEGG enrichment analysis of these differential genes , FTO regulatory pathways were studied and downstream target genes of FTO were screened . The role of FTO downstream target gene in HCC was revealed by bioinformatic analysis and cell experiments .@*Results@#Transcriptome sequencing showed that 386 genes were differentially expressed between HepG2 cells of knock⁃down FTO and HepG2 cells , and they were involved in biological processes such as response to interferon⁃gamma . The expression of IFIT2 , one of the most responsive interferon⁃stimulating genes , was up⁃regulated after FTO knockdown . Potential m6 A methylation occurred at multiple sites of IFIT2 . The survival of HCC patients with high expression of IFIT2 was significantly prolonged , and knock⁃down of IFIT2 promoted the growth and migration of HepG2 cells . @*Conclusion @#FTO may regulate IFIT2 by mediating m6 A , and further promote the occurrence and development of HCC .
摘要
ObjectiveTo study the effect of Qizhu Kang'ai prescription (QZAP) on the gluconeogenesis enzyme phosphoenolpyruvate carboxykinase 1 (PCK1) in the liver of mouse model of liver cancer induced by diethylnitrosamine (DEN) combined with carbon tetrachloride (CCl4) and Huh7 cells of human liver cancer, so as to explore the mechanism on regulating metabolic reprogramming and inhibiting cell proliferation of liver cancer cells. MethodDEN combined with CCl4 was used to construct a mouse model of liver cancer via intraperitoneal injection. A normal group, a model group, and a QZAP group were set up, in which QZAP (3.51 g·kg-1) or an equal volume of normal saline was administered daily by gavage, respectively. Serum and liver samples were collected after eight weeks of intervention. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyltransferase (γ-GT), and alpha-fetoprotein (AFP) in mice were detected to evaluate liver function changes of mice in each group. Hematoxylin-eosin (HE) staining and Sirius red staining were used to observe pathological changes in liver tissue. In the cell experiment, Huh7 cells were divided into blank group, QZAP low, medium, and high dose groups and/or PCK1 inhibitor (SKF-34288 hydrochloride) group, and Sorafenib group. The corresponding drug-containing serum and drug treatment were given, respectively. Cell counting kit-8 (CCK-8) method, colony formation experiment, Edu fluorescent labeling detection, intracellular adenosine triphosphate (ATP) content detection, and cell cycle flow cytometry detection were used to evaluate the proliferation ability, energy metabolism changes, and change in the cell cycle of Huh7 cells in each group. Western blot was used to detect the protein expression levels of PCK1, serine/threonine kinase (Akt), phosphorylated Akt (p-Akt), and cell cycle-dependent protein kinase inhibitor 1A (p21). ResultCompared with the model group, the pathological changes such as cell atypia, necrosis, and collagen fiber deposition in liver cancer tissue of mice in the QZAP group were alleviated, and the number of liver tumors was reduced (P<0.01). The serum ALT, AST, γ-GT, and AFP levels were reduced (P<0.01). At the cell level, compared with the blank group, low, medium, and high-dose groups of QZAP-containing serum and the Sorafenib group could significantly reduce the survival rate of Huh7 cells (P<0.01) and the number of positive cells with Edu labeling (P<0.01) and inhibit clonal proliferation ability (P<0.01). The QZAP groups could also reduce the intracellular ATP content (P<0.05) and increase the distribution ratio of the G0/G1 phase of the cell cycle (P<0.05) in a dose-dependent manner. Compared with the model group and blank group, PCK1 and p21 protein levels of mouse liver cancer tissue and Huh7 cells in the QZAP groups were significantly reduced (P<0.05,P<0.01), and the p-Akt protein level was significantly increased (P<0.01). Compared with the blank group, the ATP content and cell survival rate of Huh7 cells in the SKF-34288 hydrochloride group were significantly increased (P<0.05), but there was no statistical difference in the ratio of Edu-positive cells and the proportion of G0/G1 phase distribution. Compared with the SKF-34288 hydrochloride group, the QZAP combined with the SKF-34288 hydrochloride group significantly reduced the ATP content, cell survival rate, and Edu-positive cell ratio of Huh7 cells (P<0.05) and significantly increased the G0/G1 phase distribution proportion (P<0.05). ConclusionQZAP may induce the metabolic reprogramming of liver cancer cells by activating PCK1 to promote Akt/p21-mediated tumor suppression, thereby exerting an anti-hepatocellular carcinoma proliferation mechanism.
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@#Objective: To examine the inhibitory effect of Hydrangea serrata extract against hepatocellular carcinoma HepG2 cells and its underlying mechanisms. Methods: The effects of Hydrangea serrata extract on growth inhibition of tumor cells and spheroids were assessed using MTT and 3D culture assays. Quantitative real-time PCR and Western blot analyses were employed to investigate the changes in mRNA and protein expression levels of molecules related to cell cycle and apoptosis. Results: Hydrangea serrata extract effectively inhibited the growth of both tumor cells and spheroids. The extract also significantly upregulated p27 mRNA expression and downregulated CDK2 mRNA expression, leading to cell cycle arrest. Moreover, increased BAX/ Bcl-2 ratio as well as caspase-9 and -3 were observed after treatment with Hydrangea serrata extract, indicating the induction of tumor cell apoptosis. Conclusions: Hydrangea serrata extract has the potential to alleviate tumors by effectively modulating cell-cycle-related gene expressions and inducing apoptosis, thereby inhibiting tumor growth.
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Objective @# To investigate the impact of dexmedetomidine on the oncological behavior of hepatocellular carcinoma and explore the role of NF-E2-related factor 2 (Nrf2) at both in vitro and in vivo levels.@*Methods @# In vivo experiment,Male C57BL/6J mice were randomly divided into a control group ( Ctrl group) ,a hepatocellular carcinoma group ( HCC group) ,and a hepatocellular carcinoma + dexmedetomidine group ( HCC + Dex group) . Hepatocellular carcinoma was induced in mice by combining N-Nitrosodiethylamine ( DEN) / carbon tetrachloride ( CCl4 ) ,followed by daily intraperitoneal injection of 10% dexmedetomidine for two weeks.After feeding the mice for one month,the mice were assessed for the quantity and size of liver tumors.The proliferation ability of liver cancer was evaluated using Ki67 immunohistochemistry.Additionally,the expression level of Nrf2 protein in tumor tissue was measured through immunofluorescence.In vitro experiment,Hepa1-6 cells were incubated with different concentrations of dexmedetomidine (0. 1,1,5 nmol /L) for 48 hours to examine their effects.The proliferation, migration and invasion abilities of Hepa1-6 cells were evaluated using the MTT and Transwell methods.The expres- sion level of Nrf2 protein in the Hepa1-6 cells was measured using Western blot and immunofluorescence.Addition- ally,the proliferation ,migration and invasion abilities of cells were assessed after Nrf2 knockdown via si-RNA transfection,in combination with incubation with 1 nmol /L dexmedetomidine for 48 hours. @*Results @#ompared to the HCC group,the anatomical examination results revealed an increase in the number of liver tumors and the lon- gest diameter in the HCC + Dex group (P <0. 05) . Ki67 immunohistochemistry results indicated the number of Ki67 positive cells in liver cancer tissue increased in the HCC + Dex group (P<0. 01) .The immunofluorescence assay demonstrated an upregulation of Nrf2 expression level in the HCC + Dex group (P <0. 05 ) . MTT results showed that 1 nmol /L of dexmedetomidine increased the cell viability of Hepa1-6 cells (P<0. 05) .Transwell re- sults indicated that 0. 1 ,1 ,and 5 nmol /L of dexmedetomidine enhanced the invasive ability of Hepa1-6 cells, while 0. 1 and 1 nmol /L of dexmedetomidine enhanced the migration ability (P<0. 05) .Western blot and immu- nofluorescence results showed an upregulation of Nrf2 expression level in cells after treatment with 1 nmol /L dexme- detomidine (P<0. 01) .The Nrf2 expression level of cells was reduced using si-RNA,followed by treatment with 1 nmol /L dexmedetomidine.The results from MTT and Transwell assays revealed a decrease in the viability,invasion and migration ability of Hepa1-6 cells (P<0. 01) .@*Conclusion @# Dexmedetomidine may enhance the proliferation, invasion and migration capacity of hepatocellular carcinoma by upregulating the expression of Nrf2 .
摘要
Objective @#To investigate the expression , synergistic relationship and clinical significance of alcohol dehydrogenase (ADH1A) and vascular endothelial growth factor-A (VEGFA) in hepatocellular carcinoma (HCC) . @*Methods @#The expression and correlation of ADH1A and VEGFA in HCC and adjacent normal tissues were ana lyzed by GEPIA . TCGA and GSEA were used to analyze the pathway of ADH1A in HCC . The clinical and patho logical data of 84 patients with HCC were collected , and 54 patients with paracancer normal tissue samples were se lected as controls to analyze the correlation between ADH1A and VEGFA and clinicopathological parameters of HCC . Immunohistochemistry was used to detect the protein expression of ADH1A and VEGFA in cases and con trols , and the correlation between the expression of ADH1A and VEGFA and the clinical progression and prognosis of patients with HCC was analyzed based on clinical pathological parameters and Kaplan Meier.@*Results @#Bioinfor matics analysis found that ADH1A was low expressed in HCC and VEGFA was highly expressed in HCC , and there was a negative correlation between the two ( P < 0.001) ; immunohistochemical detection results showed that the expression of ADH1A in HCC tissue was lower than that in normal tissue adjacent to cancer (P < 0.01) while the expression rate of VEGFA in HCC tissue was significantly higher than that of normal tissue adjacent to cancer (P < 0.01) ; The recurrence rate of vascular thrombus and HCC patients in HCC group with high expression of ADH1A was lower (P < 0.05) . The proportion of tumor diameter > 5 cm , high TNM stage , microsatellite and G2 G3 dif ferentiation in HCC tissues in VEGFA high expression group was higher (P < 0.05) . Kaplan Meier survival analy sis showed that patients with high ADH1A expression and low VEGFA expression had a higher five year survival rate .@*Conclusion @#Low expression of ADH1A and high expression of VEGFA in tumor tissues of patients with HCC indicate tumor progression and can be used as one of the prognostic evaluation indicators for patients with HCC .
摘要
ObjectiveTo investigate the molecular mechanism by which Si Junzitang in intervening in the development of hepatocellular carcinoma (HCC) by regulating the O-linked β-N-acetylglucosamine modification (O-GlcNAcylation) of nuclear factor kappa-B (NF-κB) p65 in the paracancerous tissues. MethodThe orthotopic liver cancer mouse model was established. Twenty-four C57BL/6 mice were randomly divided into four groups: Normal group, model group, Si Junzitang low-dose group (10 g·kg-1), and Si Junzitang high-dose group (25 g·kg-1), with 6 mice in each group. The O-GlcNAcylation level and phosphorylation modification level of p65 in the paracancerous tissues were detected using Western blot. The O-GlcNAcylation of p65 was assessed using immunoprecipitation (IP). The mRNA expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), vascular endothelial growth factor A (VEGFA), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) in the paracancerous tissues was detected using real-time quantitative polymerase chain reaction (Real-time PCR). The tumor number, liver weight, locomotor activity, grip strength, and Qi status of the mice were observed and analyzed. ResultCompared with the normal group, the model group showed a significant decrease in O-GlcNAcylation in the paracancerous tissues (P<0.01), a significant decrease in p65 O-GlcNAcylation (P<0.01), a significant increase in p65 phosphorylation (P<0.01), significantly elevated mRNA levels of cytokines IL-6, TNF-α, TGF-β1, VEGFA, MMP-2, and MMP-9 (P<0.01), significantly increased liver weight (P<0.01), significantly declined grip strength, number of grid crossings, and number of vertical stand-ups (P<0.01), and significantly dwindled Qi status (P<0.01). Compared with model group, the Si Junzitang low-dose and high-dose groups showed significantly increased levels of O-GlcNAcylation in the paracancerous tissues (P<0.05, P<0.01), significantly upregulated p65 O-GlcNAcylation levels (P<0.05, P<0.01), and significantly decreased p65 phosphorylation levels (P<0.01). In the Si Junzitang low-dose group, the mRNA levels of IL-6, TGF-β1, and VEGFA significantly decreased (P<0.05, P<0.01). In the Si Junzitang high-dose group, the mRNA levels of IL-6, TNF-α, TGF-β1, VEGFA, MMP-2, and MMP-9 significantly decreased (P<0.01), the number of tumors larger than 3 mm in diameter significantly decreased (P<0.01), and liver weight significantly decreased (P<0.05). Additionally, grip strength, number of grid crossings, and number of vertical stand-ups significantly increased (P<0.05, P<0.01), along with a significant increase in qi status (P<0.01). ConclusionSi Junzitang can inhibit the progression of orthotopic HCC in mice, which may be achieved by increasing the O-GlcNAcylation level in the paracancerous tissues, enhancing the O-GlcNAcylation of p65, inhibiting the phosphorylation modification of p65, and ultimately suppressing the expression of downstream IL-6, TNF-α, TGF-β1, VEGFA, MMP-2, and MMP-9.
摘要
Hepatocellular carcinoma (HCC) is a common tumor in the digestive tract, the formation mechanism of which remains to be fully elucidated. Although surgery, radiation, chemotherapy, targeted therapy, and immunotherapy have achieved significant results in the treatment of HCC, these methods are accompanied by a considerable number of adverse reactions and complications. In recent years, Chinese medicine has shown remarkable efficacy in the treatment of HCC, and both basic experiments and clinical studies have confirmed the effectiveness of Chinese medicine, which exerts therapeutic effects via multiple components and multiple targets. However, the pathogenesis of HCC is exceptionally complex and not fully understood, which means that studies remain to be carried out regarding the specific mechanism of Chinese medicine in preventing and treating HCC. Network pharmacology and molecular biology can be employed to decipher the mechanism of Chinese medicine in the treatment of diseases. Studies have shown that Chinese medicine can regulate various pathways such as the mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), Hedgehog, Wnt/β-catenin, nuclear factor-κB (NF-κB), Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3), and transforming growth factor-β (TGF-β)/Smad signaling pathways. Chinese medicine can exhibit its anti-HCC effects by inducing cell apoptosis, inhibiting cell proliferation and migration, and blocking the cell cycle via the above pathways. However, the specific mechanisms remain to be systematically studied. This study comprehensively reviews the regulatory effects of Chinese medicine on HCC-related signaling pathways to reveal the molecular mechanisms of Chinese medicine in the treatment of HCC. This view holds the promise of providing new targets, new perspectives, and new therapies for HCC treatment and advancing the modernization and development of Chinese medicine.
摘要
Objective To analyze the current status,hotspots,and trends in the research field of traditional Chinese medicine and pharmacy combined with transcatheter arterial chemoembolization(TACE)for the treatment of hepatocellular carcinoma.Methods The relevant literatures on traditional Chinese medicine and pharmacy combined with TACE treatment for hepatocellular carcinoma collected by China National Knowledge Infrastructure from January 1994 to December 2022,and use CiteSpace 6.1.R6 software to analyze its publication volume,authors,research institutions,and key words.Results This study included a total of 686 articles.The publication volume showed a fluctuating upward trend from 1994 to 2015,but it had decreased since 2016,indicating that further breakthrough research was needed in this field.The communication between authors and research institutions needed to be strengthened,with low levels of cross institutional and cross regional cooperation and uneven geographical distribution.Cooperation should be strengthened.High frequency key words included hepatocellular carcinoma,Chinese materia medica,interventional therapy,therapeutic,etc.,resulting in 9 key word clustering labels.Conclusion Currently,there is a lack of experimental research and related mechanism research,the future research hotspots of traditional Chinese medicine and pharmacy combined with TACE treatment for hepatocellular carcinoma will focus more on traditional Chinese medicine and pharmacy,interventional therapy,clinical observation,and cell apoptosis,which provides new ideas for researchers conducting related research in the future.
摘要
Objective Microvascular invasion(MVI)risk scoring model was established based on the preoperative data of hepatocellular carcinoma(HCC)patients.Methods The clinical data of 1153 HCC patients who underwent hepatectomy in Hangzhou First People's Hospital from January 2000 to December 2021 were retrospectively analyzed.Random sampling method was used to divide the samples into modeling group(n=864)and verification group(n=289)at a ratio of 3:1.The modeling group used Logistic regression analysis model to explore the independent risk factors of MVI and established a prediction model accordingly.Receiver operating characteristic(ROC)curve and correction curve were drawn to evaluate the predictive ability and performance of the risk model.Results The incidence of MVI was 24.1%(208/864)in modeling group.Multivariate Logistic regression analysis showed that alpha-fetoprotein(AFP)>160ng/ml and total tumor volume(TTV)>30cm3 were independent risk factors for MVI(P<0.05).The total score of risk scoring model was 6 points,0-1 was classified as low risk,2-3 was classified as medium risk,and 4-6 was classified as high risk.The model predicted that the area under the curve(AUC)of MVI was 0.714 in modeling group and 0.731 in verification group.The calibration diagram showed that the prediction model had good performance.Conclusion The MVI risk prediction model for HCC patients based on TTV and AFP is simple and easy to use,which is conducive to preoperative treatment decision-making and doctor-patient communication.