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Transarterial chemoembolization （TACE） is currently the primary treatment method for advanced liver cancer. This article elaborates on the current status of application of TACE in hepatocellular carcinoma from the aspects of existing techniques， patient selection， and efficacy assessment and summarizes the research advances and prospects of TACE combined with local treatment and systemic therapy， so as to provide new ideas for clinical practice and experimental studies.
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Hepatocellular carcinoma (HCC) is malignant tumor with the fourth incidence rate and the second mortality rate in China, and patients with advanced stage have lost the chance of surgical treatment, short survival period and extremely poor prognosis. Histopathological biopsy is the gold standard for clinical diagnosis of malignant tumors, but histopathological biopsy is not only invasive, but also obtains fewer tissue samples, which does not reflect the heterogeneity of tumors, and makes it difficult to dynamically monitor the progression of tumors or the efficacy of treatment. Therefore, it is clinically important to find new non-invasive strategies for early detection of HCC and to monitor the efficacy of HCC. Circulating tumor DNA is a non-invasive liquid biopsy method with simple sampling and can dynamically monitor the genomic changes of tumors, which has great application value in early diagnosis, therapeutic efficacy monitoring, and prognostic evaluation of HCC.
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Objective To study whether emodin,a natural product,can affect the level of histone acetylation in HpG2 hepatocellular carcinoma cells(HCC),and then accelerate the occurrence of pyroptosis and apoptosis in hepatocellular carcinoma cells,so as to provide a new target for the treatment of liver cancer.Methods CCK-8 method was used to detect the effect of different concentrations of emodin on the viability of HpG2 cells.Bioinformatics was used to analyze histone acetylation-related genes in patients with liver cancer in TCGA database.The correlation between the candidate gene lysine acetyltransferase 2A(KAT2A)and the apoptosis pathway was verified.qPCR method was used to detect the mRNA level of KAT2A in HepG2 cells and L02 cells.The effects of emodin on histone acetyltransferase(HAT)and histone deacetyltransferase(HDAC),interleukin 1β(IL-1β)and interleukin 18(IL-18)in HpG2 cells were detected by ELISA.The effect of emodin on the apoptosis of liver cancer cells was detected by flow cytometry.The expression level of cell apoptosis,pyroptosis-associated protein B lymphocytoma-2(Bcl-2),Bcl-2-related X protein(Bax),NOD-like receptor thermal protein domain-related protein 3(NLRP3),Caspase-1,Gasdermin family member DN terminal(GSDMD-N)and KAT2A were detected by Western blot assay.Results Emodin could reduce the activity of HpG2 cells,and the confidence interval of IC5095%was 58.12-66.52μmol/L.Compared with normal liver tissue,the expression of histone acetylation related gene mRNA was increased in HCC tissue,and the change of KAT2A was the highest[log2(Fold Change)=2.010,P＜0.01].In HCC tissue,the expression of KAT2A mRNA was negatively correlated with apoptosis pathway(rs=-0.230,P＜0.01).Compared with L02 cells,the expression of KAT2A mRNA in HepG2 was higher(P＜0.05).Compared with the control group,expression levels of HAT and HDAC decreased in the 60μmol/L emodin intervention group,expression levels of IL-18 and IL-1β increased,the apoptosis rate increased,expression levels of KAT2A and BAX decreased,and expression levels of Bcl-2,NLRP3,GSDMD-N and Caspase-1 increased(P＜0.05).Conclusion Emodin could inhibit the viability of hepatoma cells,accelerate apoptosis and pyroptosis,and its mechanism may be related to the regulation of KAT2A expression.
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Objective:To explore the application value of laparoscopic contrast enhanced ultrasound in laparoscopic surgery for patients with hepatocellular carcinoma combined with cirrhosis.Methods:The clinical data of 71 patients with hepatocellular carcinoma combined cirrhosis from February 2018 to February 2020 in Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology were retrospectively analyzed. The patients underwent preoperative enhanced CT and multi-parameter MRI examination, followed by laparoscopic partial hepatectomy, and intraoperative laparoscopic contrast enhanced ultrasound examination. Based on histological examination and follow-up results, the diagnostic efficacy of preoperative imaging and preoperative imaging combined with intraoperative laparoscopic contrast enhanced ultrasound in patients with hepatocellular carcinoma combined with cirrhosis was compared.Results:Among the 71 patients, 69 completed laparoscopic surgery and 2 converted to open surgery. One hundred and ten HCC lesions were diagnosed by preoperative imaging examination, 105 lesions were detected by intraoperative ultrasound among them, of which 98 lesions were diagnosed as HCC by intraoperative laparoscopic contrast enhanced ultrasound. There were no statistically significant difference in sensitivity, specificity, accuracy, positive predictive value and negative predictive value between preoperative imaging and preoperative imaging combined with intraoperative laparoscopic contrast enhanced ultrasound in the diagnosis of malignant liver lesions: 94.4% (102/108) vs. 99.1% (107/108), 81.0% (34/42) vs. 66.7% (28/42), 90.6% (136/150) vs. 90.0% (135/150), 92.7% (102/110) vs. 88.4% (107/121) and 85.0% (34/40) vs. 96.6% (28/29), P>0.05. Laparoscopic contrast enhanced ultrasound revealed an additional 11 suspected malignant lesions, of which 5 lesions were histologically confirmed as HCC. Seven patients underwent surgical strategy changes. Conclusions:Laparoscopic contrast enhanced ultrasound in patients with HCC combined with cirrhosis during laparoscopic surgery can be used to detect, identify, accurately locate of the lesions and modify the surgical plan.
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Objective:Transcriptomics combined with proteomics was used to analyze the potential signaling pathways of epidermal growth factor-like domain 9 (EGFL9) affecting the proliferation, invasion and migration of hepatocellular carcinoma.Methods:RNA interference technique was used to build hepatocellular carcinoma cell line with EGFL9 Huh-7 gene knockdown, the control group (NC group) and experimental group (KD group), each group of three samples, were performed the transcriptome and proteomics analysis, screening differences genes and proteins, to express the correlation analysis, cluster analysis, and subsequently gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) were used for gene function and pathway annotation enrichment analysis, respectively.Results:Based on omics analysis, there were 8 335 different genes in KD group compared with NC group, among which 4 207 were up-regulated and 4 128 were down-regulated. There were 298 different proteins, of which 188 were up-regulated and 110 down-regulated. Based on the combined analysis of the two omics, 213 differentially expressed genes were found. Among them, the top three common differentially expressed genes at the level of transcription and translation were transferrin receptor 2 (TFR2), annexin A1 (ANXA1) and solute carrier family 38 member 2(SLC38A2). The common differentially expressed genes were significantly enriched in cell cycle signaling pathway, amino acid biosynthesis pathway, p53 signaling pathway and glycolysis/gluconeogenesis signaling pathway.Conclusion:EGFL9 may participate in the regulation of cell function of hepatocellular carcinoma cells by regulating the expression of TFR2, ANXA1, LC38A2 and other genes, and may play a role through the regulation of cell cycle and other molecular signaling pathways.
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Primary liver cancer is a malignant tumor with high morbidity and mortality. It also ranks in the forefront in the incidence and mortality of malignant tumors in China, which seriously threatens the lives and health of Chinese people. Most patients have already been in the intermediate and late stage when they are diagnosed, thus the chance of surgery is lost, and the prognosis is poor. In recent years, with the advancement of vascular interventional therapy technologies such as hepatic arterial chemoembolization and hepatic arterial infusion chemotherapy, the emergence of new tyrosine kinase inhibitors, immune checkpoint inhibitors, and especially the development of multimodal combination therapy, the treatment effect of unresectable hepatocellular carcinoma has been continuously improved, and it also provides a potential possibility for sequential surgical treatment. This article reviews the research progress of vascular interventional therapy combined with systemic therapy in unresectable hepatocellular carcinoma, in order to provide a reference for the clinical treatment of unresectable hepatocellular carcinoma.
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Objective:To evaluate the reoperation of abdominal metastasis after liver resection for hepatocellular carcinoma (HCC).Methods:The data of 2748 patients with liver cancer undergoing surgical resection at the Department of Hepatobiliary and Pancreatic Surgery of Li Huili Hospital of Ningbo Medical Center from January 2010 to January 2022 were retrospectively screened. A total of 19 patients with abdominal metastases after liver resection undergoing reoperation were enrolled, which were all males with a median age of 53 years (27 to 68). The surgical procedures and diagnosis for abdominal metastases were recorded, and the recurrence and survival of patients were followed up.Results:During the follow-ups of initial resection of HCC, 10 patients were diagnosed with postoperative abdominal metastasis by enhanced CT, and seven patients were diagnosed by MRI. MRI and PET/CT were negative in two patients. Abdominal metastasis was found during reoperation in one case and liver transplantation in the other case due to postoperative liver recurrence. All 19 patients successfully underwent radical resection of abdominal metastases. Eighteen patients underwent open surgery and one underwent laparoscopic surgery. Among them, nine cases underwent simple metastases resection, six combined liver resection, one combined liver resection and right hemicolectomy, one combined partial rectal resection, one combined partial small bowel resection, and one combined liver transplantation. The 1-year, 3-year, and 5-year disease-free survival rates were 26.3%, 15.8%, 10.5%, respectively. The 1-year, 3-year, and 5-year overall survival rates were 94.7%, 26.3%, 15.8%, respectively. Three patients are currently surviving disease-free for 154.3 months, 67.3 months, and 33.4 months, respectively. These three patients all had single abdominal metastase and did not receive any targeted or immune treatments after surgery.Conclusion:For patients with localized or single abdominal metastases after HCC surgery, reoperation for metastases can bring survival benefits.
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Objective:To evaluate the relationship between red blood cell distribution width (RDW) and prognosis of patients with hepatocellular carcinoma (HCC) andergoing transcatheter arterial chemoembolization (TACE).Methods:Clinical data of 212 patients with HCC andergoing TACE for the first time in Department of Interventional Radiology, the Affiliated Hospital of Xuzhou Medical University from January 2011 to May 2018 were retrospectively analyzed, including 184 males and 28 females, aged (56.8±11.2) years. Follow-up for survival. X-tile software was used to determine 13.1% as the optimal threshold for preoperative RDW prediction of prognosis, and enrolled patients were divided into a low level group (RDW<13.1%, n=70) and a high level group (RDW≥13.1%, n=142). Aspartate aminotransferase, total bilirubin, albumin, hemoglobin and lipoprotein a, Barcelona clinical liver cancer (BCLC) stage and other indexes were compared between the two groups. Survival analysis was performed by Kaplan-Meier method, survival rate was compared by log-rank test, and the effect of RDW on prognosis was analyzed by Cox regression. Results:The 1-year, 2-year and 3-year cumulative survival rates in RDW high level group were 34.5%, 14.1% and 6.3%, respectively, while those in RDW low level group were 64.3%, 38.6% and 21.4%, respectively, with significant difference ( χ2=23.09, P<0.001). Compared with the low level group, the levels of aspartate aminotransferase and total bilirubin were higher, the levels of albumin, hemoglobin and lipoprotein a were lower, the proportion of portal vein cancer thrombin was higher, and the stage of BCLC was later, with statistical significance (all P<0.05). Cox regression analysis showed that HCC patients with RDW≥13.1%( HR=1.732, 95% CI: 1.223-2.452, P=0.002) had poor survival prognosis after TACE. Conclusion:Preoperative RDW≥13.1% is an independent risk factor for survival after TACE in patients with HCC. RDW has potential predictive value for prognosis of patients with HCC.
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Objective:To investigate the expression, prognostic value and mechanism of MCM10 in hepatocellular carcinoma (LIHC).Methods:The transcriptome and clinical data of hepatocellular carcinoma were obtained from The Cancer Genome Atlas database, and the rank sum test was used to analyze the expression level of MCM10 in tumor tissues and adjacent tissues. Cox regression analysis was used to analyze the relationship between MCM10 expression level and the survival prognosis of patients with hepatocellular carcinoma. Gene set enrichment analysis (GSEA) was utilized for pathway enrichment analysis between MCM10 high and low group gene expression profiles. The effect of MCM10 knockdown on the proliferation of HepG2 cells was determined by cell counting kit-8 (CCK-8) cell viability assay. The effect of MCM10 knockdown on the expression of G1/S-specific cyclin D1 was detected by Western blot.Results:The expression value of MCM10 in hepatocellular carcinoma was 0.709±0.595, and that in adjacent tissues was 0.077±0.094 ( P<0.0 001). Cox regression analysis showed that high expression of MCM10 was a risk factor and prognostic predictor of overall survival ( HR=1.32, 95% CI: 1.19~1.48) and disease-specific survival ( HR=1.40, 95% CI: 1.22~1.61) in LIHC. GSEA analysis showed that the differentially expressed genes were mainly enriched in cell cycle, p53 signaling pathway and positive regulation of G1-S phase transition, et al. CCK-8 assay showed that MCM10 knockdown could inhibit the proliferation of HepG2 cells. Western blot analysis further confirmed that knockdown of MCM10 expression inhibited the expression of cyclin D1 in HepG2 cells. Conclusions:MCM10 is a risk factor for the prognosis of patients with hepatocellular carcinoma, which can promote the proliferation of hepatoma cells through cyclin D1.
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Objective:To analyze the differences between trans-radial access (TRA) and trans-femoral access (TFA) in hepatic arterial perfusion chemotherapy (HAIC) in terms of patient experience, postoperative complications, and patient preferences; explore whether TRA in HAIC is associated with better patient experience and compliance; and determine whether it is safer than TFA.Methods:The study was a retrospective cohort study of patients with advanced hepatocellular carcinoma and liver metastases from colorectal cancer treated with HAIC. We enrolled a total of 91 patients with advanced liver malignancies treated with HAIC from November 2022 to May 2023 in the Department of Interventional Therapy and Hepatobiliary Medicine at Tianjin Medical University Cancer Hospital. The patients were divided into three groups: group TRA ( n=20, receiving TRA HAIC only), group TFA ( n=33, receiving TFA HAIC only), and crossover group [ n=19, receiving TFA HAIC (Cross-TFA group) first, followed by TRA HAIC (Cross-TRA group)]. Meanwhile, to facilitate the expression of partial results, all patients receiving TRA HAIC were defined as the TRA-HAIC group ( n=39, TRA+Cross-TRA group), and all patients receiving TFA HAIC were defined as the TFA-HAIC group ( n=52, TFA+Cross-TFA group). The primary research index was the Quality of Life (QOL) visualization scale score. The secondary research index included approach-related and catheter-related adverse events, duration of surgery, and mean length of patient stay. We used various statistical methods such as Mann-Whitney U test, t-test, Chi-square test, Fisher′s exact test, univariate logistic regression analysis, and multi-factor analysis. Results:TRA patients had significantly lower QOL scores than TFA patients (all P<0.001). The QOL scores of the Cross-TRA group were significantly lower than those of the Cross-TFA group (pain at the puncture site Z=-3.24, P=0.001, others P<0.001). The QOL scores of the Cross-TRA group were compared with those of the TRA group, which showed that the scores of the Cross-TRA group in overall discomfort ( Z=-3.07, P=0.002), postoperative toilet difficulty ( Z=-2.12, P=0.034), and walking difficulty ( Z=-2.58, P=0.010) were significantly lower than those of the TRA group. Satisfaction scores were significantly higher in the Cross-TRA group than in the Cross-TFA group ( Z=-3.78, P<0.001), and patients were more likely to receive TRA HAIC as the next procedure ( χ2=30.42, P<0.001). In terms of mean length of stay, patients receiving TRA HAIC had a significantly lower mean length of stay than those receiving TFA HAIC (50.1±3.2 h vs. 58.4±6.4 h, t=7.98, P<0.001). The incidence of radial artery occlusion (RAO) as an approach-related adverse event was 15.4% (6/39) in the TRA-HAIC group, which was significantly higher than that in the TFA-HAIC group (15.4% vs. 0, χ2=8.56, P=0.005). Notably, multifactorial analysis of RAO-related factors showed that intraoperative enoxaparin use and patency of radial artery flow during pressure were significantly associated with a reduced risk of postoperative RAO ( P=0.037 for enoxaparin use and P=0.049 for pressure). Conclusions:With respect to procedure approach, TRA was significantly better than TFA in terms of patient satisfaction and mean length of stay. Through further process optimization and prevention of adverse reactions, the incidence of adverse reactions can be maintained at a relatively low level, so that patients can benefit from TRA in future operations in terms of cost-effectiveness and medical efficiency.
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Objective:To observe the expression of CD36 in hepatocellular carcinoma tissues and cell lines, and to investigate the effects of CD36 on the proliferation and migration abilities of human hepatocellular carcinoma cell lines and human hepatocellular carcinoma cell xenograft models in nude mice.Methods:Differences in the expression levels of CD36 transcripts in 371 hepatocellular carcinoma and paracancerous tissues were analyzed based on information from The Cancer Genome Atlas (TCGA) database. Cancer tissues and corresponding paracancerous tissues of 48 hepatocellular carcinoma patients who were diagnosed and underwent surgical treatment at the Affiliated Hospital of Yangzhou University from January 2019 to February 2021 were prospectively collected, and the levels of CD36 mRNA in the tissues were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) method. Western blotting was used to detect CD36 protein levels in human hepatocellular carcinoma cell lines Huh7 and HCCLM3 and human normal liver cell line LO2. Plasmids containing CD36 interfering sequences and empty plasmids were transfected into Huh7 cells or HCCLM3 cells for sh-CD36 group and control group, respectively. The CCK-8 assay was used to detect the proliferation ability (expressed as absorbance value) of cells in each group at 0, 12, 24, 36, 48 and 60 h of culture, and the scratch healing assay and Transwell assay were used to detect the migration ability of cells in each group. The Huh7 cells of sh-CD36 group or control group were injected into the axillary subcutis of BALB/c nude mice, with 4 mice in each group, to construct nude mice models of human hepatocellular carcinoma xenografts; the long and short diameters of tumor were measured weekly after 1 week of inoculation, and the tumor volume was calculated. The nude mice were put to death after 5 weeks of inoculation, and the tumor specimens were collected and weighed; the tumor cell morphology was observed under the microscope, and the expressions of CD36 and Ki-67 proteins in the tumor tissues was detected by immunohistochemistry (IHC).Results:Analysis of the data from the TCGA database showed that the level of CD36 transcripts was higher in hepatocellular carcinoma tissues compared with that in paracancerous tissues (4.2±1.8 vs. 3.2±1.5, t = 2.28, P = 0.035). Tissues detection using qRT-PCR in 48 patients with hepatocellular carcinoma showed that the relative expression of CD36 mRNA in hepatocellular carcinoma tissues was higher than that in paracancerous tissues (0.76±0.26 vs. 0.48±0.23, t = 3.52, P < 0.001). Western blotting assay showed that CD36 protein level in Huh7 and HCCLM3 cells was higher than that in LO2 cells, which were (1.42±0.11) times and (1.68±0.16) times higher than LO2 cells, respectively (both P < 0.001). At the mRNA and protein levels, the CD36 of Huh7 and HCCLM3 cells in the sh-CD36 group was lower than that in the corresponding control group (both P < 0.001). CCK-8 assay showed that the proliferative ability of Huh7 cells and HCCLM3 cells in the sh-CD36 group was lower than that in the corresponding control group after 36 and 24 h of culture (both P < 0.01). Scratch healing assay showed that the scratch healing rates of Huh7 cells [(12±3)% vs. (30±5)%, t = 4.01, P < 0.001] and HCCLM3 cells [(15±4)% vs. (29±5)%, t = 4.16, P < 0.001] in the sh-CD36 group were lower than those in the corresponding control group at 48 h of culture; Transwell assay showed that the number of Huh7 cells [(46±6) cells/field of view vs. (88± 6) cells/field of view, t = 5.56, P < 0.001] and HCCLM3 cells [(42±5) cells/field of view vs. (82±7) cells/field of view, t = 5.34, P < 0.001] penetrating into the membrane in 24 h in the sh-CD36 group was less than that in the corresponding control group. Five weeks after subcutaneous injection, the tumor volume [(682±268) mm 3vs. (1 375±512) mm 3, t = 4.73, P = 0.006] and tumor mass [(432±95) mg/mouse vs. (871±109) mg/mouse, t = 6.57, P < 0.001] of nude mice injected with Huh7 cells of the sh-CD36 group were lower than those of nude mice injected with Huh7 cells of the control group; under the microscope, the density of tumor cells in transplanted tumor specimens of nude mice injected with Huh7 cells of the sh-CD36 group was lower than that in nude mice injected with Huh7 cells of the control group, and the expression levels of both CD36 and Ki-67 proteins were also low. Conclusions:CD36 expression is up-regulated in cancer tissues of hepatocellular carcinoma patients and human hepatocellular carcinoma cell lines Huh7 and HCCLM3, and it may associate with cell proliferation and migration of hepatocellular carcinoma. Knockdown of CD36 expression significantly inhibits the proliferation and migration abilities of hepatocellular carcinoma cells in vitro, and inhibits the tumors of human hepatocellular carcinoma cell xenograft models in nude mice.
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ObjectiveTo investigate the safety and efficacy of tyrosine kinase inhibitors combined with immune checkpoint inhibitors in the treatment of patients with Child-Pugh class B unresectable hepatocellular carcinoma （uHCC）. MethodsA total of 96 patients with Child-Pugh class B uHCC who were admitted to Beijing Ditan Hospital， Capital Medical University， from December 31， 2020 to March 30， 2023 were enrolled as subjects， among whom 63 patients receiving lenvatinib combined with programmed death-1 （PD-1） inhibitor were enrolled as L group and 33 patients receiving sorafenib combined with PD-1inhibitor were enrolled as S group. The primary endpoint was objective response rate （ORR）， and secondary endpoints included time to progression （TTP）， overall survival （OS）， toxicity， drug withdrawal rate， and dose adjustment rate. The The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups； the chi-square test was used for comparison of categorical data between two groups. Survival curves were plotted， and the Kaplan-Meier method was used to calculate the survival rate of patients in both groups， while the Log-rank test was used for comparison between the two groups. The Cox regression model was used to calculate hazard ratio （HR） and its 95% confidence interval （CI） and perform the multivariate analysis of influencing factors for prognosis. ResultsAmong the 96 patients with uHCC， 55 （57.3%） had Child-Pugh class B （7 points） uHCC and 41 （42.7%） had Child-Pugh class B （8—9 points） uHCC. The L group had a significantly higher ORR than the S group （46.0% vs 15.2%， P=0.003）， and there were no significant differences between the L group and the S group in median TTP （6.6 months vs 3.5 months， P=0.48） or OS （13.8 months vs 13.2 months， P=0.95）. There was no significant difference in median TTP between the patients with Child-Pugh class B （7 points） uHCC and those with Child-Pugh class B （8—9 points） uHCC （6.6 months vs 4.8 months， P=0.35）， while there was a significant difference in OS between these two groups of patients （14.5 months vs 8.8 months， P=0.045）. The multivariate analysis showed that ORR was a protective factor for both TTP （HR=0.18， 95%CI： 0.09 — 0.36， P<0.001） and OS （HR=0.20， 95%CI： 0.09 — 0.43， P<0.001）. There were no significant differences between the L group and the S group in the overall incidence rate of adverse reactions （98.4% vs 97.0%） and the incidence rate of grade≥3 adverse reactions （68.3% vs 63.6%）， and there were also no significant differences between the two groups in dose adjustment rate （84.8% vs 70.2%） and drug withdrawal rate （56.1% vs 72.7%）. ConclusionCompared with the regimen of sorafenib combined with PD-1 inhibitor， the regimen of lenvatinib combined with PD-1 inhibitor can improve the ORR of patients with Child-Pugh class B uHCC， with similar prognosis and safety profile between the two groups.
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ObjectiveTo investigate the expression of essential meiotic endonuclease 1 （EME1） in liver cancer tissue and its effect on the biological behavior of hepatoma cells. MethodsThe TCGA database was used to identify the differentially expressed genes between liver cancer tissue and paracancerous tissue. Immunohistochemistry and Western Blot were used to measure the expression abundance of EME1 in liver cancer tissue. A lentivirus was constructed by short hairpin RNA， and BEL-7404 cells were transfected with the lentivirus to interfere with the expression of the EME1 gene； the cells were divided into silencing group （shEME1 group） and control group （shCtrl group）. Quantitative real-time PCR and Western Blot were used to measure the mRNA and protein expression levels of EME1； Celigo Image Cytometer and MTT assay were used to measure cell proliferation rate； flow cytometry was used to observe cell cycle； Caspase 3/7 activity was used to measure cell apoptosis. The independent-samples t-test was used for comparison between two groups. ResultsTCGA results showed that the mRNA expression level of EME1 in liver cancer tissue was 18.9 times that in paracancerous tissue （t=5.00， P<0.001）， and the protein expression level of EME1 in liver cancer tissue was 7.0 times （based on immunohistochemistry： 8.4±2.6 vs 1.2±0.4， t=7.55， P<0.001） or 2.5 times （based on Western Blot： 249.0%±35.5% vs 100.0%±77.8%， t=3.02， P<0.05） that in paracancerous tissue. After lentivirus infection， compared with the shCtrl group， the shEME1 group had an mRNA expression level of EME1 reduced by 29.9% （29.9%±0.9% vs 100.0%±3.6%， t=32.82， P<0.001）， a protein expression level of EME1 reduced by 35.7% （35.7%±14.9% vs 100.0%±28.9%， t=3.42， P<0.05）， and a level of cell counting reduced by 45.1% （4 053±167 vs 8 988±477， t=16.91， P<0.001）， as well as a level of cell activity reduced to 66.9% （0.518±0.046 vs 0.774±0.022， t=8.74， P<0.001） and a level of colony forming ability reduced to 29.0% （75±6 vs 260±9， t=28.92， P<0.001）. Compared with the shCtrl group， the shEME1 group had a significant increase in the proportion of cells in G1 phase （49.9% vs 44.0%， t=8.96， P<0.001） and significant reductions in the proportion of cells in G2/M phase （15.9% vs 17.9%， t=9.13， P<0.001） and S phase （34.2% vs 38.1%， t=6.91， P<0.001）， while Caspase 3/7 activity was enhanced by 1.5 times （145.8%±5.9% vs 100.0%±2.3%， t=12.50， P<0.001）. ConclusionEME1 is highly expressed in liver cancer tissue， and silencing the EME1 gene can inhibit the proliferation of hepatoma cells and promote cell apoptosis.
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ObjectiveTo investigate the effect of ankyrin-repeat domain-containing protein 22 （ANKRD22） on the proliferation， invasion， and migration of human hepatoma cells and its molecular mechanism. MethodsThe TCGA database was used to analyze the expression level of ANKRD22 in normal liver tissue and hepatocellular carcinoma tissue and its association with prognosis. Western Blot and qRT-PCR were used to measure the expression of ANKRD22 in human normal liver cells （L-02） and human hepatoma cells （Huh7， HepG2， MHCC-97H， SK-HEP-1， and SMMC-7721）； CCK-8 assay， EdU， wound healing assay， and Transwell assay were used to observe the effect of ANKRD22 on the proliferation， invasion， and migration of hepatoma cells； Western Blot was used to investigate the association of ANKRD22 with cyclins and EMT-related proteins； KEGG and ssGSEA analyses were performed to investigate the mechanism of action of ANKRD22 in hepatoma cells， and related experiments were conducted for validation. The independent-samples t-test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsIn the TCGA database， the expression level of ANKRD22 in hepatoma tissue was significantly higher than that in normal liver tissue （t=5.083， P<0.05）， and the patients with a high expression level of ANKRD22 had longer overall survival and disease-related survival than those with a low expression level of ANKRD22 （P<0.05）. The expression level of ANKRD22 in various human hepatoma cell lines was higher than that in human normal liver cells （all P<0.05）. Cell proliferation assay showed that the ANKRD22 overexpression group had significantly higher EdU positive rate and proliferation rate than the Vector group （t=19.60 and 6.72， both P<0.001）， and compared with the si-NC group， the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower EdU positive rate and proliferation rate （all P<0.001）. Compared with the Vector group， the overexpression group had significantly higher expression levels of Cyclin E1， Cyclin D1， CDK7， and CDK4 （t=3.54， 4.95， 6.34， and 5.19， all P<0.01）， and the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower expression levels than the si-NC group （all P<0.001）. The overexpression group had a significantly lower expression level of P27 than the Vector group （t=6.12， P<0.001）， and the si-ANKRD22#2 group and the si-ANKRD22#3 group had a significantly higher expression level than the si-NC group （both P<0.001）. Invasion and migration experiments showed that compared with the Vector group， the ANKRD22 overexpression group had significantly higher migration rate and number of crossings through the membrane （migration group and invasion group） （t=5.01， 25.60， and 3.67， all P<0.05）， and compared with the si-NC group， thesi-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower migration rate and number of crossings through the membrane （migration group and invasion group） （all P<0.01）. The overexpression group had significantly higher expression levels of N-cadherin， Vimentin， and Snail than the Vector group （t=12.13， 8.85， and 13.97， all P<0.001）， and the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower expression levels than the si-NC group （all P<0.001）； the overexpression group had a significantly lower expression level of E-cadherin than the Vector group （t=4.98， P<0.01）， and the si-ANKRD22#2 group and the si-ANKRD22#3 group had a significantly higher expression level than the si-NC group （both P<0.001）. The KEGG enrichment analysis and the ssGSEA analysis showed that ANKRD22 was associated with the PI3K/AKT/mTOR signaling pathway in hepatocellular carcinoma， and the overexpression group had significantly higher expression levels of p-AKT/AKT， p-PI3K/PI3K， and p-mTOR/mTOR than the Vector group （t=12.21， 3.43， and 9.75， all P<0.01）， and the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower expression levels than the si-NC group （all P<0.001）. ConclusionANKRD22 is highly expressed in hepatoma cells and can promote the proliferation， invasion， and migration of hepatoma cells and the activation of the PI3K/AKT/mTOR signaling pathway.
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Liver macrophages are important immune cells in the liver， and they express proinflammatory factors and anti-inflammatory factors through polarization into M1 type and M2 type， respectively， thereby playing a role in regulating inflammatory damage response. The malignant transformation of hepatic progenitor cells is the core mechanism of the malignant progression of hepatic precancerous lesions， and its key factor is the continuous stimulation of inflammatory microenvironment， which is closely associated with M1/M2 macrophage polarization. This review mainly focuses on the association between macrophage polarization， chronic inflammation， and malignant transformation of hepatic progenitor cells， so as to provide a theoretical basis for the prevention and treatment of hepatic precancerous lesions.
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ObjectiveTo investigate the efficacy and safety of cryoablation combined with Camrelizumab monoclonal antibody in the treatment of hepatocellular carcinoma （HCC）. MethodsA total of 103 HCC patients who were admitted to our hospital from June 2020 to June 2023 were enrolled and randomly divided into combined treatment group with 53 patients and control group with 50 patients. The patients in the control group received percutaneous argon-helium cryoablation， and those in the combined treatment group received percutaneous argon-helium cryoablation combined with Camrelizumab monoclonal antibody. The two groups were compared in terms of short-term response， changes in T lymphocyte subsets after treatment， changes in liver function and alpha-fetoprotein （AFP） after treatment， and progression-free survival and overall survival during follow-up. The t-test was used for comparison of normally distributed continuous data between groups， and the chi-square test was used for comparison of categorical data between groups. The Kaplan-Meier method was used to plot survival curves， and the log-rank test was used for comparison of survival time between the two groups. ResultsThe combined treatment group had significantly higher overall response rate and disease control rate than the control group （χ2=4.156 and 4.348， P=0.042 and 0.037）. After treatment， the combined treatment group had significant increases in the percentages of CD3+ and CD4+ T lymphocytes and CD4+/CD8+ ratio （P<0.05） and a significant reduction in the percentage of CD8+ T lymphocytes （P<0.05）， while the control group had no significant changes in T lymphocyte subsets after treatment （P>0.05）， and compared with the control group after treatment， the combined treatment group had significantly higher percentages of CD3+ and CD4+ T lymphocytes and CD4+/CD8+ ratio （all P<0.05） and a significantly lower percentage of CD8+ T lymphocytes （P<0.05）. After treatment， both groups had significant reductions in the levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and AFP （all P<0.05） and a significant increase in the level of albumin （Alb） （P>0.05）， and compared with the control group after treatment， the combined treatment group had significantly lower levels of ALT， AST， and AFP （all P<0.05） and a significantly higher level of Alb （P<0.05）. There were no significant differences in the incidence rates of grade Ⅲ—Ⅳ （moderate to severe） adverse reactions between the two groups （P>0.05）. Compared with the control group， the combined treatment group had significantly better median progression-free survival （21.32 months vs 15.31 months， χ2=4.689， P=0.030） and median overall survival （28.36 months vs 20.75 months， χ2=5.030， P=0.025）. ConclusionArgon-helium cryoablation combined with Camrelizumab monoclonal antibody can effectively improve short-term response， enhance immune function， and prolong survival time， with a favorable safety profile.
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Hepatocellular carcinoma （HCC） is the most common type of primary liver cancer and the third leading cause of cancer-related deaths， and it is a serious threat to human health and has become a clinical problem that needs to be solved urgently. Extracellular vesicles （EV） are membrane vesicles containing multiple components and play an important role in the development and progression of HCC. This article summarizes the effect of EVs of different origins on HCC and analyzes the mechanism of action of EV on HCC， so as to provide new perspectives for the diagnosis and treatment of HCC.
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ObjectiveTo investigate whether anti-PD-1 monoclonal antibody can improve the efficacy and safety of cryoablation combined with lenvatinib in the treatment of unresectable hepatocellular carcinoma （HCC）. MethodsA retrospective analysis was performed for 232 patients with unresectable HCC who were treated at The Fifth Medical Center of Chinese PLA General Hospital from January 2018 to December 2022， among whom 128 received cryoablation combined with lenvatinib （double combination） and 104 received cryoablation combined with lenvatinib and anti-PD-1 monoclonal antibody （triple combination）. Propensity score matching was performed at a ratio of 1∶1， and finally there were 86 patients in each group. The two groups were evaluated in terms of objective response rate （ORR）， disease control rate （DCR）， overall survival （OS）， progression-free survival （PFS）， and adverse events （AEs）. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups； the chi-square test was used for comparison of categorical data between two groups. Survival curves were plotted， and the Kaplan-Meier method was used to calculate the survival rate of patients in both groups， while the log-rank test was used for comparison between the two groups. The Cox regression model was used to calculate hazard ratio （HR） and 95% confidence interval （CI） and perform the univariate and multivariate analyses of influencing factors for prognosis. ResultsThe median follow-up time was 28 months， and there were 33 deaths （38.0%） in the triple combination group and 40 deaths （46.0%） in the double combination group. Compared with the double combination group， the triple combination group had significantly higher ORR （35.6% vs 14.5%， P=0.008） and DCR （86.1% vs 64.1%， P=0.003）. OS and PFS in the triple combination group were significantly higher than those in the double combination group （P=0.045 and 0.026）. The univariate and multivariate Cox proportional-hazards regression model analyses showed that treatment regimen （HR=0.60， P=0.038） and alpha-fetoprotein level （HR=2.37， P=0.001） were independent risk factors for OS， and treatment regimen （HR=0.65， P=0.025）， diabetes mellitus （HR=1.94， P=0.005）， whether or not to have received local treatment （HR=0.63， P=0.014）， and distant metastasis （HR=0.58， P=0.009） were independent risk factors for PFS. There was no significant difference in the incidence rate of AEs between the two groups （P>0.05）. ConclusionFor patients with unresectable HCC， the triple combination of cryoablation， lenvatinib， and anti-PD-1 monoclonal antibody significantly improves the treatment outcome and survival of patients compared with the double combination of cryoablation and lenvatinib， without increasing AEs， which provides a clinical basis for optimizing the treatment regimen for unresectable HCC.
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ObjectiveTo evaluate the efficacy and safety of first-line transcatheter arterial chemoembolization （TACE） combined with targeted therapy and immunotherapy in the treatment of patients with stage Ⅱb/Ⅲa hepatocellular carcinoma （HCC） based on China Liver Cancer Staging （CNLC）. MethodsA total of 198 patients who received first-line TACE combined with targeted therapy and immunotherapy or received TACE alone from January 2015 to December 2022 in the First Affiliated Hospital of Soochow University were enrolled in this study， and after propensity score matching， there were 50 patients in combination group and 50 patients in TACE group. The Kaplan-Meier method was used to calculate median overall survival （mOS） and median progression-free survival （mPFS）. Modified Response Evaluation Criteria in Solid Tumors was used to evaluate objective response rate （ORR） and disease control rate （DCR）， and Common Terminology Criteria for Adverse Events v5.0 was used to evaluate adverse events. The chi-square test was used for comparison of categorical data between two groups； the t-test was used for comparison of normally distributed continuous data between two groups， and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. The Kaplan-Meier method was used to estimate survival time and calculate 95% confidence interval （CI）， and the Log-rank test was used for comparison of mOS and mPFS between two groups. ResultsThe combination group had an mOS of 30.1 months （95%CI： 21.9 — 38.3）， and the TACE group had an mOS of 14.5 months （95%CI： 11.0 — 18.0）， with a significant difference between the two groups （χ2=17.8， P<0.001）； the combination group had an mPFS of 10.3 months （95%CI： 8.8 — 11.8）， and the TACE group had an mPFS of 7.1 months （95%CI： 5.8 — 8.4）， with a significant difference between the two groups （χ2=10.4， P<0.001）. There were significant differences between the combination group and the TACE group in ORR （84% vs 58%， P<0.05） and DCR （94% vs 80%， P<0.05）. There was no significant difference between the combination group and the TACE group in the incidence rate of adverse events （24% vs 16%， P=0.317）， and no adverse event-related deaths were observed in either group. ConclusionCompared with TACE alone， TACE combined with targeted therapy and immunotherapy has a better efficacy in the treatment of patients with CNLC stage Ⅱb/Ⅲa HCC， without increasing the incidence rate of severe adverse events.
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In recent years， transcatheter arterial chemoembolization （TACE） has emerged as a common treatment modality for the treatment of hepatocellular carcinoma （HCC）. However， with the ongoing development of embolic agent techniques， the new advances in microspheres and nanoparticles have brought new hope for improving the efficacy and safety of TACE. This article reviews the latest advances and applications of microspheres and nanoparticles in TACE for HCC. First， this article introduces the background of TACE as a therapeutic approach and the emergence of microsphere and nanoparticle techniques， and then it describes the application of various types of microspheres and nanoparticles in TACE and discusses the requisite attributes of an ideal embolic agents. The article focuses on the advances in material science and engineering， as well as the clinical efficacy of drug-eluting microspheres and nanoparticles versus conventional TACE. Furthermore， it discusses the importance of radiological examination in TACE and summarizes the research advances in the radiopaque and magnetic resonance-visible embolic agents. This article also explores the future development directions and challenges of TACE. It also points out the combination of microspheres and nanoparticles with other treatment modalities， the application of personalized and precision medicine in TACE， and the potential regimen of TACE in clinical translation， and meanwhile， it raises the issues of ethics and regulation that need to be further discussed. It is believed that microspheres and nanoparticles have a potential effect in TACE， which provides a theoretical basis and technical support for innovating HCC treatment regimens and improving the prognosis of patients through TACE interventions.