【 Abstract 】 Objective To probe into the role of inositol 1, 4, 5-trisphosphate (IP3) and bax gene expression in apoptosis of HepG2 cells induced by genistein (Gen). Methods HepG2 cells were treated with different concentrations including 20, 40, 60 and 80 μ mol/L Gen as HepG2 cells cultured with 0 μmol/L Gen for 72 h was control; HepG2 cells were treated with 60 μmol/L Gen for 6, 12, 24, 48 and 72 h as HepG2 cells treated with 60 μmol/L Gen for 0 h was control. IP3 content, bax mRNA expression and apoptosis rate were assayed by IP3- ［ 3H ］ Birtrak assay, RT-PCR and flow cytometry, respectively. ResultsHepG2 cells incubated with each concentration of Gen for 72 h ， IP3 content was lower than that of control 〔 (17.7 ± 1.3), (11.2 ± 0.9), (4.9 ± 0.5), (4.8 ± 0.3) pmol/106 cells vs (29.4 ± 0.5) pmol/106 cells 〕 , P ＜ 0.01 ； bax mRNA expression (RI which was the gray degree multiply area of bax/the gray degree multiply area of β -actin) was higher than that of control (0.26 ± 0.02, 0.33 ± 0.05, 0.35 ± 0.06, 0.38 ± 0.05 vs 0.09 ± 0.01), P ＜ 0.01 ； The apoptosis rate was higher than that of control 〔 (10.1 ± 0.9)%, (18.7 ± 1.6)%, (28.7 ± 2.5)%, (27.9 ± 2.0)% vs (2.6 ± 0.1)% 〕 , P ＜ 0.01. HepG2 cells were incubated with 60 μ mol/L Gen for 6, 12, 24, 48 and 72 h ， IP3 content was lower than that of control 〔 (22.6 ± 0.9), (12.0 ± 1.4), (7.5 ± 0.8), (5.6 ± 0.5), (4.3 ± 0.6) pmol/106 cells vs (29.2 ± 0.6) pmol/106 cells 〕 , P ＜ 0.01 ； bax mRNA expression was higher than that of control incubated with 60 μ mol/L Gen for above 12 h (0.25 ± 0.06, 0.29 ± 0.02, 0.30 ± 0.02, 0.35 ± 0.04 vs 0.09 ± 0.01), P ＜ 0.01 ； The apoptosis rate in groups incubated with 60 μ mol/L Gen for 24, 48 and 72 h was significantly higher than that in control 〔 (7.4 ± 0.5)%, (20.5 ± 2.0)%, (30.7 ± 1.6)% vs (2.6 ± 0.1)% 〕 , P ＜ 0.01. ConclusionGen induces apoptosis of HepG2 cells by reducing IP3 production and increasing bax gene expression.
ObjectiveTo investigate the expression and clinical significance of cytochromes b561 (CYB561) in hepatocellular carcinoma (HCC). MethodsThe expression of CYB561 mRNA in HCC tissues and its relationship with prognosis were analyzed by database data. Immunohistochemistry (IHC) was used to detect the expression of CYB561 protein in 61 matched HCC tissues and their adjacent tissues, and the relationship between CYB561 protein expression and clinicopathological features and prognosis of HCC was analyzed. Kaplan-Meier method was used to draw the survival curve and Cox proportional hazard regression model was used to analyze the correlation between the expression of CYB561 protein and the prognosis of HCC. ResultsThe analysis of database data showed that the relative expression of CYB561 mRNA in HCC tissues was higher than that in adjacent tissues (P<0.001). Compared with HCC patients with negative expression of CYB561 mRNA, HCC patients with positive expression of CYB561 mRNA had worse overall survival (OS), relapse-free survival, progression-free survival and disease-free survival (all P<0.05). The results of IHC showed that the positive rates of CYB561 protein in HCC tissues and adjacent tissues were 57.38% (35/61) and 21.31%(13/61), respectively. The former was higher than the latter, with statistical significance (χ2=16.624, P<0.001). Survival analysis showed that the OS of patients with positive expression of CYB561 protein was worse than that of patients with negative expression (P<0.05). Multivariate Cox proportional hazard regression analysis showed that the positive expression of CYB561 protein was a risk factor for postoperative OS in HCC patients [HR=3.308, 95%CI (1.344, 8.144), P=0.009]. ConclusionCYB561 is positively expressed in HCC and suggests a worse survival, and may serve as a potential prognostic biomarker for HCC.
Objective To explore the aberrantly expressed genes in hepatocellular carcinoma (HCC) and their relationship with prognosis of HCC through bioinformatics analysis. Methods Five datasets related to HCC were selected from the GeneExpression Omnibus database to explore differentially expressed genes (DEGs), followed by further gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The co-upregulated genes CNIH4 and TOMM40 were selected to explore the differences in their expressions in HCC tissues and normal tissues, and to explore the relationship between their expressions and the 5-year survival of patients by using TCGA database. Tissues and paraneoplastic tissues of eight cases of HCC who underwent surgery at the Guangdong Second Provincial General Hospital were collected to verify the expression differences of CNIH4 and TOMM40L mRNA. Results A total of 25 up-regulated genes and 21 down-regulated genes were identified in this study. The results of GO analysis and KEGG analysis indicated that DEGs were mainly related to catabolism, cell division, DNA replication and repair. The results of TCGA database analysis showed that the expression of up-regulated genes CNIH4 mRNA and TOMM40L mRNA were up-regulated in HCC tissues as compared with normal tissues (P<0.05) and that the 5-year survival of patients in the high expression group was worse than that in the low expression group (P<0.05). The results of clinical samples showed that CNIH4 mRNA and TOMM40L mRNA were up-regulated in HCC tissues as compared with paraneoplastic tissues. Conclusion CNIH4 and TOMM40L genes are up-regulated in HCC tissues, and their high expressions are associated with poor prognosis, and may be potential biomarkers and prognostic indicators for HCC.
ObjectiveTo investigate the value of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid enhanced magnetic resonance imaging (Gd-EOB-DTPA-MRI) in gross morphological classification of hepatocellular carcinoma (HCC). MethodsThe clinicopathologic data of patients with HCC who received surgical treatment in the Affiliated Huai’an Hospital of Xuzhou Medical University from January 2017 to December 2022 were retrospectively gathered. The Gd-EOB-DTPA-MRI was performed before operation. Two radiologists independently assessed the gross morphological classification of HCC according to the imaging performance. The tumors were cut into sections in a coronal plane and were taken pictures for recording pathological features after operation. The tumors were assigned into 4 types according to the references and clinical experiences: single nodular type (SN), single nodular with extranodular growth type (SN-EG), confluent multi-nodular type (CMN), and infiltration type (IF). Matching degree of morphological classification was analyzed between by the Gd-EOB-DTPA-MRI and resected specimen. The pathological features of 4 types of HCC were also analyzed. ResultsA total of 87 patients with HCC were included. The gross morphological classification by the Gd-EOB-DTPA-MRI was 28 (32.2%) patients with SN, 28 (32.2%) patients with SN-EG, 21 (24.1%) patients with CMN, 10 (11.5%) patients with IF, which by the resected specimen was 33 (37.9%) patients with SN, 24 (27.6%) patients with SN-EG, 21 (24.1%) patients with CMN, and 9 (10.4%) patients with IF in the 87 patients with HCC. The Kappa’s coefficient of agreement between the results of Gd-EOB-DTPA-MRI and postoperative resection specimens was 0.776 (P=0.199). There were statistical differences in the tumor diameter and microvascular invasion (MVI) among the 4 types of gross morphology classification (F=2.937, P=0.038; χ2=16.852, P=0.001), the MVI rate was highest and tumor diameter was biggest in the patients with IF among the 4 types of gross morphology classification (P<0.05). ConclusionsFrom the results of this study, the gross morphological classification of HCC is closely related to the tumor diameter and MVI. Results of Gd-EOB-DTPA-MRI and postoperative resection specimens in assessing the gross morphological classification are good agreement. Therefore, an accurate preoperative planning and better therapy strategy for the patients with HCC can be provided according to gross morphological classification by preoperative Gd-EOB-DTPA-MRI.
ObjectiveTo introduce the basic principles of commonly used assessment methods for liver function reserve, and compare the advantages and disadvantages of various assessment methods, so as to provide a reference for hepatectomy of patients with hepatocellular carcinoma (HCC). MethodThe literature on evaluation methods of liver reserve function in patients with HCC at home and abroad in recent years was searched and summarized. ResultsFrom the results of literature review, the Child‐Pugh score and indocyanine green discharge test were the most commonly used to assess preoperative liver function reserve for patients with HCC. The application value of other examinations such as albumin-bilirubin score, gadolinium-ethoxybenzyl diethylenetriamine pentaacetic acid-enhanced magnetic resonance imaging (Gd-EOB-DTPA-enhanced MRI), nuclear medical imaging in predicting post-hepatectomy liver failure was gradually being explored. ConclusionsThe combination of clinical parameters and volumetric studies is used to assess preoperative liver function reserve for patients with HCC. The clinical applications of nuclear medical imaging and Gd-EOB-DTPA-enhanced MRI make up for the deficiency of local liver function reserve evaluation, which are important examinations to assess liver function reserve after conversion therapy in the future. However, more domestic studies are still needed to confirm their values.
With the publication of a vast amount of clinical research on hepatocellular carcinoma (HCC), the American Association for the Study of Liver Diseases (AASLD), the National Comprehensive Cancer Network (NCCN), the American Society of Clinical Oncology (ASCO), and the National Health Commission of China have all updated their diagnostic and treatment guidelines for HCC. There are no differences in the definition of HCC risk populations among the AASLD 2023, NCCN 2024, and China Liver Cancer Staging and Treatment Guideline (CNLC) 2024. Notably, CNLC 2024 has updated its guidance on high-risk factors and prospective surveillance for HCC based on the characteristics of HCC patients in China. The four guidelines have seen significant updates in the areas of neoadjuvant and adjuvant therapies, local treatments, and systemic treatments for HCC. CNLC 2024 refines the indications for local treatment, improves systemic treatment, and introduces new first-line therapy, including camrelizumab combined with rivoceranib or tislelizumab. The second-line therapy nivolumab plus ipilimumab for advanced HCC are recommended by AASLD 2023, NCCN 2024, and ASCO 2024, which may become a new first-line therapeutic option for patients with advanced HCC. We compare and interpret these four guidelines in this paper.
ObjectiveTo exclusively compare the short-and long-term outcomes of hepatic resection (HR) patients with multifocal tumors meeting the Milan criteria between locating in same and different sections. MethodsA total of 219 consecutive HR patients with multifocal tumors meeting the Milan criteria were divided into group SS (n=97, same section) and group DS (n=122, different sections) according to their anatomical location (Couinaud's segmentation). ResultsThe 1-, 3-, and 5-year overall survival (OS) and recurrence-free survival (RFS) rates were significantly higher in the group SS than those in the group DS (P < 0.05). The subgroup analysis showed that patients with 2 tumors and those undergoing en bloc resection were associated with better OS and RFS (P < 0.05). ConclusionsFor HCC patients with multifocal tumors meeting the Milan criteria, those with tumors locating in same hepatic section may have better longterm survival and lower HCC recurrence rates than those locating in different sections after HR.
ObjectiveTo summarize the application of circulating free DNA (cfDNA) in the diagnosis and treatment of hepatocellular carcinoma (HCC). MethodThe relevant literature on the application of cfDNA in the diagnosis and treatment of HCC both domestic and international was reviewed and summarized. ResultsThe cfDNA is an emerging biomarker in recent years. At present, the different detection methods had been reported in a large number of studies to detect abnormal methylation, hot spot mutation, gene copy number variation, quantitative detection of cfDNA concentration, etc. It was found that the cfDNA could be used in the management process of early diagnosis, treatment guidance, and efficacy evaluation of HCC patients. ConclusionscfDNA detection is a good tool in the diagnosis and treatment of HCC, which can help clinicians make-decisions and bring more possibilities for the diagnosis and treatment of HCC, which is of great significance for changing the current diagnosis and treatment of HCC. However, there are still many challenges in cost control, technology optimization, and standardization of evaluation indicators. With the continuous progress of molecular biology technology and artificial intelligence, the application of cfDNA in diagnosis and treatment of HCC will be further expanded, its advantages will be better played, and the related shortcomings will be gradually solved.
Objective To dynamically study the formation of multidrug resistance(MDR) of human hepatocellular carcinoma cell SMMC-7721 induced by Adriamycin (ADM) and the role of multidrug resistance-associated protein(MRP) in its mechanisms.Methods Hepatocellular carcinoma cell SMMC-7721 was cultured in RPMI-1640 medium containing ADM with progressively increased concentration or directly cultured in medium containing different concentrations of ADM. Resistant index of drug-resistant variants of SMMC-7721 cell was determined by drawing cell dosage-reaction curves.Levels of MRP mRNA expression were detected by reverse transcription-polymerase chain reaction(RTPCR). Intracellular rubidomycin(DNR) concentration was examined by flow cytometry(FCM).Results With progressive increasing of ADM concentration in medium resistant index and levels of MRP mRNA expression were correspondingly increased but intracellular DNR concentration was markly reduced. When parental cells were directly cultured in medium containing different concentrations of ADM, the higher the ADM concentration, the higher the level of MRP mRNA expression, but intracellular DNR concentration was kept at the similar high level and most cells died. Conclusion ADM may progressively induce SMMC-7721 cell resistant to multiple chemotherapeutic drugs with reduced intracellular DNR accumulation associated with the overexpression of MRP gene.