The treatment of intermediate and advanced-stage hepatocellular carcinoma poses significant challenges, prompting a research focus on the effective evaluation and enhancement of therapies. Transarterial chemoembolization (TACE) combined with immunotherapy has exhibited potential, however, there is a need for further validation of its efficacy and the identification of biomarkers that can predict patient prognosis.
To explore the clinical value of serum vascular endothelial growth factor (VEGF) in evaluating the clinical efficacy of TACE alone or combined with target immunotherapy in advanced hepatocellular carcinoma.
The clinical data of 113 newly diagnosed patients with advanced hepatocellular carcinoma were hospitalized in the Third Hospital of Hebei Medical University from January 2021 to July 2022 were analyzed. According to the treatment regimen, the patients were divided into TACE group (n=66), TACE combined targeting group (n=22), and TACE combined with target immunity group (n=25). Tumor markers such as VEGF, alpha fetoprotein (AFP), and protein induced by vitamin K antagonist-Ⅱ (PIVKA-Ⅱ) were detected before and after treatment. According to the modified response evaluation criteria in solid tumors (mRECIST), patients were followed up for 3, 6, and 12 months after treatment to evaluate the clinical efficacy of advanced hepatocellular carcinoma. The objective response rate (ORR) and disease control rate (DCR) of the patients were analyzed. The median progression free survival (PFS) was calculated by the Kaplan-Meier and the survival curve was drawn. The receiver operating characteristic (ROC) curve was used to analyze the predictive value of serum VEGF and related combined serum tumor markers for the clinical efficacy of advanced hepatocellular carcinoma.
The ORRs of the TACE group, TACE combined targeting group and TACE combined target immunity group were after 12 months of follow-up were 17.14% (6/35), 33.33% (4/12), and 54.55% (6/11), respectively, and the DCRs were 28.57% (10/35), 41.67% (5/12), and 72.73% (8/11), respectively. The ORR and DCR of the TACE combined target immunity group after 12 months of treatment were significantly higher than those of the TACE group, with statistically significant differences (P<0.05). The median PFS of TACE combined with target immunity group (15.039 months) was better than that of TACE group (8.757 months) and TACE combined targeting group (9.680 months) (P<0.05). The frequency of TACE in TACE combined with target immunity group after 12 months of follow-up was significantly less than that in TACE groupand TACE combined targeting group (P<0.05). The difference of serum VEGF before and after treatment in TACE combined targeting group and TACE combined target immunity group was significantly higher than that in TACE group, and the difference was statistically significant (P<0.05). The AUC of the difference in serum VEGF before and after treatment for predicting the clinical efficacy of advanced hepatocellular carcinoma was 0.748 (P<0.001), and the sensitivity and specificity were 0.909 and 0.629, respectively. The AUCs of serum VEGF combined with PIVKA-Ⅱ, VEGF combined with AFP, and VEGF combined with AFP and PIVKA-Ⅱ for predicting the clinical efficacy of advanced hepatocellular carcinoma were 0.781, 0.869, and 0.872, respectively (P<0.001) .
TACE combined with targeted immunotherapy can improve the efficacy of patients with advanced hepatocellular carcinoma and prolong the PFS of patients. Serum VEGF in patients can be used as a biological indicator for evaluating clinical efficacy.
Hepatocellular carcinoma (HCC) is the leading cause of cancer-related deaths. The current prevention and treatment situation remains critical. It is of scientific significance to explore new therapeutic agents for HCC.
To analyze the mechanism of wogonin on HCC by network pharmacology and to verify it in vitro.
The drug targets of wogonin were searched in TCMSP database, and the disease targets of HCC were collected from TTD, GenCard, OMIM, DisGent databases. The collected drug targets and disease targets were intersected as potential targets for drug intervention in diseases. R software was used for enrichment analysis of intersection targets, STRING database and Cytoscape software were used to construct protein interaction network and screen core targets. The core targets were further analyzed in GIEPA database. Finally, the preliminary analysis results were verified by in vitro experiments, including cell activity determination using CCK-8 kit, cell proliferation determination using plate clone formation experiment, cell migration determination using scoring test, protein expression level determination using Western-blotting (WB) assay.
The AMDE characteristics of wogonin were found to be in accordance with the rules for small molecule drug formation and the toxicity analysis showed no toxicity. A total of 135 wogonin targets and 8 238 HCC targets were collected, and 113 targets were intersected. Through the analysis of the core genes of TOP10 screened by the constructed protein interaction network, it was found that the mRNA levels of CDK1 and SRC in liver cancer tissues were higher than those in normal liver tissues (P<0.05), and the high expression levels in liver cancer patients were related to poor prognosis (P<0.05). KEGG enrichment analysis showed that the intersection genes were enriched in the PI3K/AKT signaling pathway, and the molecular docking results showed that wogonin had strong binding configuration activity with CDK1 and SRC. The results of CCK-8 kit showed that the activity of HepG2 cells in the 75.0, 150.0, and 300.0 μmol/L wogonin groups was lower than that in the control group (P<0.05). The results of plate clone formation experiment showed that the number of colony formation of HepG2 cells in the 37.5, 75.0, 150.0 μmol/L wogonin groups was lower than that in the control group (P<0.05). The results of scoring test showed that the migration rate of HepG2 cells in the 37.5, 75.0 and 150.0 μmol/L wogonin groups was lower than that in the control group (P<0.05). The results of the WB assay showed that the expression levels of PI3K, P-AKT/AKT, CDK1 and SRC proteins in the 75.0 and 150.0 μmol/L wogonin groups were lower than those in the control group (P<0.05) .
Wogonin inhibits the proliferation and migration of HCC cells and induces apoptosis by down-regulating the expression of CDK1 and SRC proteins and attenuating the PI3K/AKT pathway signaling, to achieve the purpose of interfering with the occurrence and progression of HCC.
Hepatocellular carcinoma (HCC) is the third leading cause of common cancer-related mortality globally, accounting for approximately 90% of all primary liver cancer cases. Its recurrence and mortality rates are high, with the underlying molecular mechanisms remaining unclear.
To explore potential molecular mechanisms of HCC and explore novel biomarkers.
RNA-seq expression data and clinical information were retrieved from TCGA database, differential gene expression analysis was conducted between normal liver tissue and HCC tissue. Enrichment analysis on the differentially expressed genes was performed. Based on the gene expression data profiles of HCC in TCGA, a co-expression network was established using the WGCNA R package, and weighted gene co-expression network analysis (WGCNA) was performed to select clinically significant modules and screen candidate Hub genes; the candidate Hub genes were further analyzed for significant differential expression in HCC tissues and normal liver tissues, and whether they were significantly correlated with the overall survival and disease-free survival of HCC patients. The Hub genes were conclusively identified, and their protein expression was validated through the Human Protein Atlas database.
The genetic expression data in this study were obtained from 50 normal liver tissue samples and 373 HCC tissue samples. Through differential gene expression analysis, a total of 7 230 genes differential expression between HCC and normal hepatic tissue, comprising 3 691 up-regulated genes and 3 539 down-regulated genes in HCC were identified. Enrichment analysis showed that the up-regulated differentially expressed genes were mainly involved in cell cycle regulation and mitotic processes; the down-regulated differentially expressed genes were mainly involved in processes such as small molecule metabolism and organic acid metabolism. WGCNA identified 19 gene modules related to the clinical features of HCC patients, the cyan and purple modules were screened by analyzing the relationship between the modules and the clinical features. The first two genes in the cyan module genes that were strongly associated with both overall survival and disease-free survival of patients were VPS45 and FAM189B. In the purple module genes, first two genes that were strongly associated with both overall survival and disease-free survival of patients were CLEC1B and FCN3, respectively; therefore, VPS45, FAM189B, CLEC1B and FCN3 were identified as the final Hub genes. Immunohistochemical staining in the Human Protein Atlas database showed that VPS45 and FAM189B were expressed higher in HCC tissues than in normal liver tissues. FCN3 was expressed in HCC tissues lower than in normal liver tissues, the difference in the expression of CLEC1B between HCC tissues and normal liver tissues was not obvious.
VPS45, FAM189B, CLEC1B and FCN3 have been preliminary identified as possible novel potential biomarkers for HCC, which may provide a theoretical basis for targeted therapy of HCC.
Hepatocellular carcinoma (HCC) is the most common malignant liver tumor, with an increasing incidence rate. Since alpha-fetoprotein (AFP), the traditional serum marker for HCC, has a low sensitivity, there is a critical need for novel molecular biomarkers to enable early detection of HCC.
To detect the protein expression level of serum polyadenosine diphosphate ribose polymerase 2 (PARP2) in HCC patients in the blood of HCC patients and investigate its potential as a diagnostic marker for HCC.
PARP2 mRNA levels of 50 healthy individuals and 371 HCC patients were analyzed in the TCGA database, and the diagnostic efficacy was analyzed by plotting the receiver operating characteristic (ROC) curve of subjects diagnosed with HCC by PARP2 expression. The levels of PARP2 mRNA and protein expression were assessed in both HCC cells and normal hepatocytes. Serum samples from 38 newly diagnosed HCC patients and 38 healthy individuals undergoing physical examinations at the First Affiliated Hospital of Xinjiang Medical University from March 2021 to July 2022 were collected to measure serum PARP2 protein levels by using the enzyme-linked immunosorbent assay (ELISA) method, and their correlation with HCC clinical characteristics was analyzed. Additionally, the characteristics of serum PARP2 expression levels in diagnosing HCC, particularly in alpha-fetoprotein (AFP) -negative HCC (AFP <20 μg/L) was analyzed, and the efficacy of the combination of serum PARP2 and AFP for the diagnosis of HCC in patients with HCC and healthy individuals was evaluated.
TCGA data showed that PARP2 mRNA expression is higher in malignant tissues compared to paracancerous tissues based on the big data analysis of TCGA (P<0.001). PARP2 mRNA and protein expression levels were higher in HCC cells HepG2 compared to normal hepatocytes WRL68 (P<0.05). HCC patients had higher serum PARP2 protein expression levels compared to healthy individuals (P<0.001). Comparison of serum PARP2 expression levels in those with different lymphatic metastases and tumor counts showed statistically significant differences (P<0.05). The area under the ROC curve (AUC) of serum PARP2 expression level plotted for the diagnosis of HCC was 0.92, with a sensitivity of 76.32%, a specificity of 97.37% and a cut-off value of 19.45 μg/L. Upon serum AFP testing, 21 of the 38 HCC patients were AFP-negative HCC. The AUC of serum PARP2 protein level for diagnosing AFP-negative HCC was 0.95 (95%CI=0.88-1.00), with a sensitivity of 85.71%, a specificity of 97.37%, and a cutoff value of 19.59 μg/L. The diagnostic efficacy of PARP2 in combination with AFP was further evaluated using a "parallel" co-diagnostic approach, the results showed that the diagnostic efficacy of the combined diagnosis of HCC was 92.11%, the specificity was 94.74%, and the AUC was 0.934 2. For AFP-negative HCC patients, the sensitivity of the combined diagnosis was 85.71%, the specificity was 94.74%, and the AUC was 0.902 3.
PARP2 is highly expressed in HCC and can be used as a biological marker for HCC screening, especially in AFP-negative HCC.
