Ginsenoside Rg_3 inhibit hepatocellular carcinoma growth via intrinsic apoptotic pathway

AIM:To investigate the anti-tumor function of ginsenoside Rg3 on hepatocellular carcinoma(HCC) in vitro and in vivo,and its mechanism.METHODS:Hep1-6 and HepG2 cells were treated by Rg3 in different concentrations(0,50,100 and 200 μg/mL) in vitro.After incubation for 0,6,12,24 and 48 h,cell viability was measured by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide assay.Apoptosis was identified by terminal deoxynucleotidyl transferasemediated dUTP-biotin nick end labeling.Caspase-3 activity was measured by chromophore p-nitroanilide and flow cytometry.Bcl-2 family proteins were ascertained by Western-blotting.Mitochondria membrane potentialwas detected by 5,5',6' 6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide.Forty liver tumor-bearing C57Bl6 mice were divided randomly into 4 groups for intra-tumor injection of saline,ginsenoside Rg3,cyclophosphamide(CTX) and ginsenoside Rg3 + CTX combination.RESULTS:The survival time was followed up to 102 d.The mice in the Rg3 + CTX group showed significant increased survival time compared with those in the control group(P < 0.05).Rg3 could inhibit HCC cell proliferation and induce cell apoptosis in vitro in the concentration and time dependent manner.It also induced mitochondria membrane potential to decrease.Caspase-3 activation can be blocked by the inhibitor z-DEVD-FMK.Bax was up-regulated while Bcl-2 and Bcl-XL were down-regulated after Rg3 treatment.CONCLUSION:Our data suggested that Rg3 alone or combined with CTX inhibited tumor growth in vivo and prolonged mouse survival time by inducing HCC cell apoptosis via intrinsic pathway by expression alterations of Bcl-2 family proteins.

Ginsenoside Rk3 is a novel PI3K/AKT-targeting therapeutics agent that regulates autophagy and apoptosis in hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is the third leading cause of cancer death worldwide.Ginsenoside Rk3,an important and rare saponin in heat-treated ginseng,is generated from Rg1 and has a smaller molecular weight.However,the anti-HCC efficacy and mechanisms of ginsenoside Rk3 have not yet been characterized.Here,we investigated the mechanism by which ginsenoside Rk3,a tetracyclic triterpenoid rare ginsenoside,inhibits the growth of HCC.We first explored the possible potential targets of Rk3 through network pharmacology.Both in vitro(HepG2 and HCC-LM3 cells)and in vivo(primary liver cancer mice and HCC-LM3 subcutaneous tumor-bearing mice)studies revealed that Rk3 significantly inhibits the proliferation of HCC.Meanwhile,Rk3 blocked the cell cycle in HCC at the G1 phase and induced autophagy and apoptosis in HCC.Further proteomics and siRNA experiments showed that Rk3 regulates the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway to inhibit HCC growth,which was validated by molecular docking and surface plasmon resonance.In conclusion,we report the discovery that ginsenoside Rk3 binds to PI3K/AKT and promotes autophagy and apoptosis in HCC.Our data strongly support the translation of ginsenoside Rk3 into novel PI3K/AKT-targeting therapeutics for HCC treatment with low toxic side effects.

Modified albumin-bilirubin predicted survival of unresectable hepatocellular carcinoma patients treated with immunotherapy

BACKGROUND Modified albumin-bilirubin(mALBI)grade has been established as a survival determinant in hepatocellular carcinoma(HCC)patients who receive locoregional and targeted therapies.AIM To investigate whether mALBI could predict survival in unresectable HCC(uHCC)patients who were treated with atezolizumab plus bevacizumab(AB).METHODS A single-center,retrospective cohort study enrolled uHCC patients who received AB treatment between September 2020 and April 2023 and were followed up until June 2023.An association between mALBI and patient survival was determined using Cox proportional hazards analysis.RESULTS Of the 83 patients,67 patients(80.7%)were male with the mean age of 60.6 years.Among them,22 patients(26.5%)were classified as Barcelona Clinic Liver Cancer B,and 61 patients(73.5%)were classified as Barcelona Clinic Liver Cancer C.Cirrhosis was present in 76 patients(91.6%),with 58 patients classified as Child-Turcotte-Pugh(CTP)A and 18 as CTP B.The median overall survival(OS)and progression-free survival were 13.0 mo[95%confidence interval(CI):5.2-20.8]and 9.0 mo(95%CI:5.0-13.0),respectively.The patients were divided into two groups based on mALBI grades:42 patients(50.6%)in the mALBI 1+2a group;and 41 patients(49.4%)in the mALBI 2b+3 group.During the median follow-up period of 7.0 mo,the mALBI 1+2a group exhibited significantly better survival compared to the mALBI 2b+3 group,with a median OS that was not reached vs 3.0 mo(95%CI:0.1-6.0,P<0.001).In a subgroup of patients with CTP A,the mALBI 1+2a group also showed significantly longer survival compared to the mALBI 2b+3 group,with a median OS that was not reached vs 6.0 mo(95%CI:3.4-8.6,P<0.001).In the multivariate analysis,both CTP class and mALBI grade were independently associated with survival,with adjusted hazard ratios(95%CI)of 2.63(1.19-5.78,P=0.020)and 3.90(1.71-8.90,P=0.001),respectively.CONCLUSION mALBI grades can determine survival of uHCC patients receiving AB treatment,particularly those who have mildly impaired liver function.This highlights the importance of assessing mALBI before initiating AB treatment to optimize therapeutic efficacy in clinical practice.

challenges of advanced hepatocellular carcinoma

Hepatocellular carcinoma(HCC) is an aggressive malignancy,resulting as the third cause of death by cancer each year. The management of patients with HCC is complex,as both the tumour stage and any underlying liver disease must be considered conjointly. Although surveillance by imaging,clinical and biochemical parameters is routinely performed,a lot of patients suffering from cirrhosis have an advanced stage HCC at the first diagnosis. Advanced stage HCC includes heterogeneous groups of patients with different clinical condition and radiological features and sorafenib is the only approved treatment according to Barcelona Clinic Liver Cancer. Since the introduction of sorafenib in clinical practice,several phase Ⅲ clinical trials have failed to demonstrate any superiority over sorafenib in the frontline setting. Locoregional therapies have also been tested as first line treatment,but their role in advanced HCC is still matter of debate. No single agent or combination therapies have been shown to impact outcomes after sorafenib failure. Therefore this review will focus on the range of experimental therapeutics for patients with advanced HCC and highlights the successes and failures of these treatments as well as areas for future development. Specifics such as dose limiting toxicity and safety profile in patients with liver dysfunction related to the underlying chronic liver disease should be considered when developing therapies in HCC. Finally,robust validated and reproducible surrogate end-points as well as predictive biomarkers should be defined in future randomized trials.

Advanced hepatocellular carcinoma and sorafenib: Diagnosis, indications, clinical and radiological follow-up

Advanced stage hepatocellular carcinoma(HCC) is a category of disease defined by radiological, clinical and hepatic function parameters, comprehending a wide range of patients with different general conditions. The main therapeutic option is represented by sorafenibtreatment, a multi-kinase inhibitor with anti-proliferative and anti-angiogenic effect. Trans-arterial Radio Embolization also represents a promising new approach to intermediate/advanced HCC. Post-marketing clinical studies showed that only a portion of patients actually benefits from sorafenib treatment, and an even smaller percentage of patients treated shows partial/complete response on follow-up examinations, up against relevant costs and an incidence of drug related adverse effects. Although the treatment with sorafenib has shown a significant increase in mean overall survival in different studies, only a part of patients actually shows real benefits, while the incidence of drug related significant adverse effects and the economic costs are relatively high. Moreover, only a small percentage of patients also shows a response in terms of lesion dimensions reduction. Being able to properly differentiate patients who are responding to the therapy from non-responders as early as possible is then still difficult and could be a pivotal challenge for the future; in fact it could spare several patients a therapy often difficult to bear, directing them to other second line treatments(many of which are at the moment still under investigation). For this reason, some supplemental criteria to be added to the standard modified Response Evaluation Criteria in Solid Tumors evaluation are being searched for. In particular, finding some parameters(cellular density, perfusion grade and enhancement rate) able to predict the sensitivity of the lesions to anti-angiogenic agents could help in stratifying patients in terms of treatment responsiveness before the beginning of the therapy itself, or in the first weeks of sorafenib treatment. This would bring a strongly desirable help in clinical managements of these patients.

Evaluation of the relationship between hepatocellular carcinoma location and transarterial chemoembolization efficacy

AIM To evaluate the relationship between the location of hepatocellular carcinoma(HCC) and the efficacy of transarterial chemoembolization(TACE).METHODS We evaluated 115 patients(127 nodules), excluding recurrent nodules, treated with TACE between January 2011 and June 2014. TACE efficacy was evaluated according to m RECIST. The HCC location coefficient was calculated as the distance from the central portal portion to the HCC center(mm)/liver diameter(mm) on multiplanar reconstruction images rendered(MPR) to visualize bifurcation of the right and left branches of the portal vein and HCC center. The HCC location coefficient was compared between complete response(CR) and non-CR groups in Child-Pugh grade A and B patients.RESULTS The median location coefficient of HCC among all nodules, the right lobe, and the medial segment was significantly higher in the CR group than in the non-CR group in the Child-Pugh grade A patients(0.82 vs 0.62, P < 0.001; 0.71 vs 0.59, P < 0.01; 0.81 vs 0.49, P < 0.05, respectively). However, there was no significant difference in the median location coefficient of the HCC in the lateral segment between in the CR and in the non-CR groups(0.67 vs 0.65, P > 0.05). On the other hand, in the Child-Pugh grade B patients, the HCC median location coefficient in each lobe and segment was not significantly different between in the CR and in the non-CR groups.CONCLUSION Improved TACE efficacy may be obtained for HCC in the peripheral zone of the right lobe and the medial segment in Child-Pugh grade A patients.

Transarterial radioembolization for hepatocellular carcinoma:An update and perspectives

In the last decade trans-arterial radioembolization has given promising results in the treatment of patients with intermediate or advanced stage hepatocellular carcinoma(HCC),both in terms of disease control and tolerability profile.This technique consists of the selective intra-arterial administration of microspheres loaded with a radioactive compound(usually Yttrium90),and exerts its therapeutic effect through the radiation carried by these microspheres.A careful and meticulous selection of patients is crucial before performing the radioembolization to correctly perform the procedure and reduce the incidence of complications.Radioembolization is a technically complex and expensive technique,which has only recently entered clinical practice and is supported by scant results from phase Ⅲ clinical trials.Nevertheless,it may represent a valid alternative to transarterial chemoembolization(TACE) in the treatment of intermediate-stage HCC patients,as shown by a comparative retrospective assessment that reported a longer time to progression,but not of overall survival,and a more favorable safety profile for radioembolization.In addition,this treatment has reported a higher percentage of tumor shrinkage,if compared to TACE,for pre-transplant downsizing and it represents a promising therapeutic option in patients with large extent of disease and insufficient residual liver volume who are not immediately eligible for surgery.Radioembolization might also be a suitable companion to sorafenib in advanced HCC or it can be used as a potential alternative to this treatment in patients who are not responding or do not tolerate sorafenib.

Modified Child-Pugh grade vs albumin-bilirubin grade for predicting prognosis of hepatocellular carcinoma patients after hepatectomy

BACKGROUND Hepatectomy is the main treatment for patients with hepatocellular carcinoma(HCC)and it has a high possibility for long-term cure potential.But the postoperative mortality and recurrence rates remain high.Since the long-term prognosis of HCC patients is strongly linked to liver function,preoperative assessment of liver function is very important for HCC patients.AIM To compare the predictive power of the modified Child-Pugh(MCP)and albumin-bilirubin(ALBI)grades for the long-term outcome of HCC.METHODS From January 2010 to June 2017,a total of 204 patients with HCC who underwent surgery at the Second Affiliated Hospital of Chongqing Medical University were enrolled in this retrospective study.Multivariate Cox regression analysis was used to determine the independent predictive factors of survival and relapse.The area under the curve(AUC)was used to evaluate the discriminative performance of the MCP grade and ALBI grade to predict the postoperative overall survival(OS)time and recurrence-free survival(RFS)time.RESULTS The median OS and RFS times were 44.0 mo(range:22.0-74.0 mo)and 22.0 mo(range:5.0-45.0 mo),respectively.The median OS and RFS times of MCP grades 1,2,and 3 patients were 60.0,39.0,and 18.0 mo(P<0.001)and 36.0,15.0,and 7.0 mo(P<0.001),respectively.The median OS and RFS times of ALBI grades 1,2,and 3 patients were 56.0,26.0,and 6.0 mo(P<0.001)and 25.0,10.0,and 3.0 mo(P=0.003),respectively.Both the MCP and ALBI grades were more accurate than the Child-Pugh grade for predicting long-term prognosis.Further analysis demonstrated that for both predicting OS and RFS,the MCP grade performed better than the ALBI grade(AUC:0.642 vs 0.605 for OS;0.659 vs 0.594 for RFS).CONCLUSION The MCP grade is more accurate than the ALBI grade for predicting long-term outcome of patients with HCC.

Relationship between microvessel count and post-hepatectomy survival in patients with hepatocellular carcinoma

AIM: To elucidate the relationship between the microvessel count (MVC) by CD34 analyzed by immunohistochemical method and prognosis in hepatocellular carcinoma (HCC) patients who underwent hepatectomy based on our preliminary study. METHODS: We examined relationships between MVC and clinicopathological factors in 128 HCC patients. The modifi ed Japan Integrated Staging score (mJIS) was applied to examine subsets of HCC patients. RESULTS: Median MVC was 178/mm^2, which was used as a cut-off value. MVC was not signif icantly associated with any clinicopathologic factors or postoperative recurrent rate. Lower MVC was associated with poor disease-free and overall survivals by univariate analysis (P = 0.039 and P = 0.087, respectively) and lower MVC represented an independent poor prognostic factor in disease-free survival by Cox’s multivariateanalysis (risk ratio, 1.64; P = 0.024), in addition to tumor size, vascular invasion, macroscopic fi nding and hepatic dysfunction. Signifi cant differences in disease-free and overall survivals by MVC were observed in HCC patients with mJIS 2 (P = 0.046 and P = 0.0014, respectively), but not in those with other scores. CONCLUSION: Tumor MVC appears to offer a useful prognostic marker of HCC patient survival, particularly in HCC patients with mJIS 2.

How to assess the efficacy or failure of targeted therapy:deciding when to stop sorafenib in hepatocellular carcinoma

Sorafenib is thus far the only systemic treatment for hepatocellular carcinoma(HCC) based on the results of two randomized controlled trials performed in Western and in Eastern countries, despite a poor response rate(from 2% to 3.3%) following conventional evaluation criteria. It is now recognized that the criteria(European Association of the Study of the Liver criteria, modified response evaluation criteria in solid tumors) based on contrast enhanced techniques(computed tomography scan, magnetic resonance imaging) aimed to assess the evolution of the viable part of the tumor(hypervascularized on arterial phase) are of major interest to determine the efficacy of sorafenib and of most antiangiogenic drugs in patients with HCC. The role of alphafetoprotein serum levels remains unclear. In 2016, in accordance with the SHARP and the Asia-Pacific trials, sorafenib must be stopped when tolerance is poor despite dose adaptation or in cases of radiological and symptomatic progression. This approach will be different in cases of available second-line therapy trials. Some recent data(in renal cell carcinoma) revealed that despite progression in patients who received sorafenib, this drug can still decrease tumor progression compared to drug cessation. Then, before deciding to continue sorafenib post-progression or shift to another drug, knowing other parameters of post-progression survival(Child-Pugh class, Barcelona Clinic Liver Cancer, alphafetoprotein, post-progression patterns in particular, the development of extrahepatic metastases and of portal vein thrombosis) will be of major importance.

Selection of treatment modality for hepatocellular carcinoma according to the modified Japan Integrated Staging score

AIM： To compare the prognosis of patients who underwent hepatectomy and ablation using the modified Japan Integrated Staging score （mJIS）. METHODS： We examined the clinicopathologic records and patient outcomes in 278 HCC patients including 226 undergoing hepatectomy and 52 undergoing ablation therapy. RESULTS： Cirrhosis was more frequent in the ablation group. Tumor size, number and presence of vascular invasion were significantly higher in the operation group compared to the ablation group. The local recurrence rate adjacent to treated lesions was significantly higher in the ablation group compared to the operation group （P 〈 0.05）. The 3- and 5-year survival rates in the ablation and the operation group were 66% and 78%, and 50% and 63%, respectively, but not significantly different. Over 50% survival rates were observed in patients with a m.lIS score of 0-2 in both groups. However, survival rates with a score of 3-5 in both groups were significantly lower. CONCLUSION： According to the mJIS system, both local treatments could be selected for patients with a score of 0-2. However, for patients with a score more than 3, liver transplantation might be a better option in patients with HCC.

Assessment of circulating levels of microRNA-326,microRNA-424,and microRNA-511 as biomarkers for hepatocellular carcinoma in Egyptians

BACKGROUND Hepatocellular carcinoma(HCC)is the fifth most common cancer.Differential expression of microRNAs(miRNAs)-326,miRNA-424,and miRNA-511 has been associated with the diagnosis and prognosis of HCC in different populations.However,limited information is available regarding their expression in Egyptian HCC patients.AIM To assess the role of circulating miRNAs-326,miRNA-424,and miRNA-511 in Egyptian HCC patients.METHODS This prospective observational study included 70 HCC patients and 25 healthy controls.The circulating levels of these three miRNAs were evaluated by real-time PCR.Receiver operating characteristic curve analysis was used to test the diagnostic accuracy of micro RNA expression levels.RESULTS All miRNAs were differentially expressed in HCC patients;miRNAs326 and miRNA-424 were upregulated,while miRNA-511 was downregulated.Both miRNA-326 and miRNA-424 showed sensitivity and specificity of 97%,71.4%,and 52%,60%,respectively,to differentiate HCC from controls.Moreover,miRNA-326 was associated with survival and could differentiate between Child grades(A vs B);miRNA-424 significantly differentiated early vs intermediate stages of HCC;while miRNA-511 was significantly correlated with response to modified Response Evaluation Criteria in Solid Tumors(m RECIST).CONCLUSION We conclude that miRNA-326,miRNA-424,and miRNA-511 have diagnostic and prognostic roles in Egyptian patients with hepatitis C virus-related HCC and should be considered for better disease management.

Lenvatinib for large hepatocellular carcinomas with portal trunk invasion:Two case reports

BACKGROUND In a phase III trial of lenvatinib as first-line treatment for advanced unresectable hepatocellular carcinoma(uHCC),the drug proved non-inferior to sorafenib in terms of the overall survival,but offered better progression-free survival.However,the effects of lenvatinib in uHCC patients with a tumor thrombus in the main portal vein and/or a high tumor burden(tumor occupancy more than 50%of the total liver volume),remain unclear,because these were set as exclusion criteria in the aforementioned trial.CASE SUMMARY A 53-year-old man(case 1)and 66-year-old woman(case 2)with uHCC presented to us with a tumor thrombus in both the main portal vein and inferior vena cava,a high tumor burden accompanied by a tumor diameter greater than>100 mm,and distant metastasis,with the residual liver function classified as grade 2A according to the modified Albumin–Bilirubin grading.We started both patients on lenvatinib.The therapeutic effect,as evaluated by the modified Response Evaluation Criteria in Solid Tumors,was rated as partial response in both case 1 and case 2(at 8 wk and 4 wk after the start of lenvatinib administration,respectively).The therapeutic effect was sustained for 6 mo in case 1 and 20 mo in case 2.Fever occurred as an adverse event in both case 1 and 2,and hyperthyroidism and thrombocytopenia in only case 2,neither of which,however,necessitated treatment discontinuation.CONCLUSION Even in hepatocellular carcinoma patients with poor prognostic factors,if the liver function is well-preserved,lenvatinib is effective and safe.

Ginsenoside Rh1 regulates the immune microenvironment of hepatocellular carcinoma via the glucocorticoid receptor

Objective:Ginsenoside Rh1(G-Rh1)has been confirmed to inhibit the growth of breast cancer and colon cancer,but its therapeutic effect on hepatocellular carcinoma(HCC)is unclear.This study investigates the therapeutic effect of G-Rh1 on HCC as well as the underlying mechanism.Methods:Bioinformatics methods were used to analyze glucocorticoid receptor(GR)expression and the tumor microenvironment in HCC tissues from HCC patients.The effect of G-Rh1 on HCC cells was investigated in vitro using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.The therapeutic effect of G-Rh1 was investigated in vivo using subcutaneous transplantation models in C57BL/6J and nude mice.Additionally,the proportion of infiltrating immune cells in tumors was analyzed using flow cytometry,the GR and major histocompatibility complex class-I(MHC-I)expression of HCC cells after G-Rh1 treatment was analyzed using Western blotting,and G-Rh1-treated Hepa1-6 cells were cocultured with bone marrow-derived dendritic cells and B3Z T cells to further analyze the ability of G-Rh1 to induce dendritic cell(DC)maturation and CD8+T cell activation.Results:GR expression was upregulated in HCC tissues,and high GR expression was associated with a worsened immune microenvironment.In vitro studies showed that G-Rh1 had no significant effect on the proliferation of HCC cells,while in vivo studies showed that G-Rh1 exerted antitumor effects in C57BL/6J mice but not in nude mice.Further research revealed that G-Rh1 ameliorated the immunosuppressive tumor microenvironment,thereby enhancing the antitumor effects of lenvatinib by increasing the infiltration of CD8+T cells,mature DCs,and MHC-I-positive cells.MHC-I was upregulated by G-Rh1 via GR suppression.Moreover,overexpression of GR abolished the G-Rh1-mediated promotion of MHC-I expression in Huh7 cells,as well as the maturation of DCs and the activation of CD8+T cells.Conclusion:G-Rh1 can regulate the immune microenvironment of HCC by targeting GR,thus increasing the antitumor effect of lenvatinib.

Short-term effects of modified ultrasonography in laparoscopic anatomical hepatectomy for hepatocellular carcinoma

Background and aims:Laparoscopic hepatectomy is challenging,and ultrasound guidance is an effective aid but lacks standardization.This study aimed to evaluate a modified approach for laparoscopic ultrasonography to enhance surgical outcomes.Methods:Between January 2020 and August 2023,122 patients who underwent real-time ultrasound-guided laparoscopic anatomical hepatectomy for hepatocellular carcinoma were enrolled and divided into modified and traditional ultrasonography groups.The modified ultrasound application comprised intraoperative protocol-based laparoscopic ultrasonography comprising application scenarios;standardized positions for the surgeon,trocar,and probe;and the resulting standardized sections for various laparoscopic liver resections.Clinical characteristics and perioperative outcomes were compared between the two groups.Subgroup analysis was performed and comprised techniques for modified duct structure identification and portal vein branch puncture;both techniques were used in fluorescence probe-mounted laparoscopic liver resection using negative and positive staining procedures,respectively.Results:The traditional and modified groups comprised 64 and 58 patients,respectively.The patients’background characteristics were not significantly different between the groups.Surgical duration(283.4 min vs.225.1 min;p<0.001),Pringle maneuver duration(47.4 min vs.39.5 min;p?0.014),bleeding volume(258.6 mL vs.174.8 mL;p?0.005),overall complication rate(31.3%vs.15.5%;p?0.041),and postoperative stay were significantly greater in the traditional vs.modified ultrasonography groups,respectively.The modified method positively affected the number of punctures,success rate of staining,intraoperative bleeding volume,and operation duration.Conclusions:Modified ultrasonography improves the safety and effectiveness of laparoscopic hepatectomy.Ultrasonography is pivotal,especially in fluorescence probe-assisted laparoscopic liver resection.

Role of Pittsburgh Modified TNM Criteria in prognosis prediction of liver transplantation for hepatocellular carcinoma

Background Pittsburgh modified TNM criteria is one of the prognostic models of orthotopic liver transplantation （OLT） for hepatocellular carcinoma （HCC）. In this study, we applied this prognostic system in a series of HCC patients receiving OLT to verify its reliability in the clinical prognostic prediction. Methods The clinical record and follow-up data of 102 patients with HCC underwent OLT was collected. The patients were classified by 3 staging systems： the Pittsburgh Modified TNM Criteria, International Union Against Cancer （UICC） pTNM Staging System, and Milan Criteria. Survival rates of the patients were analyzed using the Kaplan-Meier method and the Log-Rank test, and then the prognostic values of the 3 staging systems were compared. Results Among the 3 staging systems, the Pittsburgh Modified TNM Criteria showed the best stratification of patients with different prognosis. The overall survival rates of the patients at the Pittsburgh modified TNM stage Ⅰ, Ⅱ, Ⅲ, and Ⅳ were 94.4%, 83.3%, 58.2%, and 36.8% at 1 year, and 79.4%, 62.5%, 26.2%, and 10.5% at 3 years, respectively. For those patients exceeding the Milan Criteria, the patients at Pittsburgh stages I and II had a significant higher survival rate than those at Pittsburgh stages III and IV （P〈0.001）. Conclusions The Pittsburgh Modified TNM Criteria is a more reliable postoperative staging system than the UICC pTNM staging system for HCC patients receiving OLT. As providing more accurate prognostic classification, it could be reasonable to combine the Milan Criteria for recipient selection.

Sorafenib delivery nanoplatform based on superpara- magnetic iron oxide nanoparticles magnetically targets hepatocellular carcinoma

Currently, sorafenib is the only systemic therapy capable of increasing overall survival of hepatocellular carcinoma patients. Unfortunately, its side effects, particularly its overall toxicity, limit the therapeutic response that can be achieved. Superparamagnetic iron oxide nanoparticles （SPIONs） are very attractive for drug delivery because they can be targeted to specific sites in the body through application of a magnetic field, thus improving intratumoral accumulation and reducing adverse effects. Here, nanoformulations based on polyethylene glycol modified phospholipid micelles, loaded with both SPIONs and sorafenib, were successfully prepared and thoroughly investigated by complementary techniques. This nanovector system provided effective drug delivery, had an average hydrodynamic diameter of about 125 nm, had good stability in aqueous medium, and allowed controlled drug loading. Magnetic analysis allowed accurate determination of the amount of SPIONs embedded in each micelle. An in vitro system was designed to test whether the SPION micelles can be efficiently held using a magnetic field under typical flow conditions found in the human liver. Human hepatocellular carcinoma （HepG2） cells were selected as an in vitro system to evaluate tumor cell targeting efficacy of the superparamagnetic micelles loaded with sorafenib. These experiments demonstrated that this delivery platform is able to enhance sorafenib＇s antitumor effectiveness by magnetic targeting. The magnetic nanovectors described here represent promising candidates for targeting specific hepatic tumor sites, where selective release of sorafenib can improve its efficacy and safety profile.

Diagnostic imaging for hepatocellular carcinoma

Hepatocellular carcinoma(HCC)occurs mostly in individuals with cirrhosis,which is why the guidelines of the most important scientific societies indicate that these patients are included in surveillance programs through the repetition of an ultrasound examination every 6 months.The aim is to achieve early identification of the neoplasia in order to increase the possibility of curative therapies(liver transplantation,surgery or local ablative therapies)and to increase patient survival.HCC nodules arising in cirrhotic livers show characteristic angiographic behavior that can be evaluated with dynamic multidetector computed tomography and dynamic magnetic resonance imaging(MRI).However,the use of these techniques in real life is often hindered by the lack of uniform terminology in reporting and in the interpretation of the exams reflected in the impossibility of comparing examinations performed in different centers and/or at different times.Liver Imaging Reporting and Data System?was created to standardize reporting and data collection of computed tomography and MRI for HCC.In some cases HCC arises in patients with healthy livers and,although there is evidence that angiographic behavior is not different from cirrhotic patients in this clinical situation,the guidelines still indicate the execution of a biopsy.Frequent use of palliative therapeutic techniques such as transarterial chemoembolization,transarterial radioembolization or administration of antiangiogenic drugs(sorafenib)poses problems of interpretation of the therapeutic response with repercussions on the subsequent choices that have been attempted to resolve with the use of stringent criteria such as Modified Response Evaluation Criteria In Solid Tumors.

IL-7的诱导表达增强靶向GPC3 CAR-T细胞的增殖及体外抗肿瘤活性

目的:探讨IL-7的诱导表达对靶向磷脂酰肌醇蛋白聚糖3(GPC3)嵌合抗原受体基因修饰T淋巴细胞(CAR-T细胞)的增殖和体外抗肿瘤活性的影响。方法:通过无缝克隆将GPC3 CAR序列片段插入GV400载体的Bam HⅠ/Eco RⅠ位置,构建第二代CAR慢病毒载体GPC3-BBZ及GPC3-BBZ-NFAT-IL-7,以293T细胞包装相应的慢病毒载体后,感染人T细胞制备CAR-T细胞。实验分为未转导T细胞(NT)组、GPC3-BBZ CAR-T细胞组、GPC3-BBZ-NFAT-IL-7 CAR-T细胞组。采用流式细胞术检测各组CAR-T细胞中CAR的表达水平,qPCR法检测经GPC3蛋白激活的CAR-T细胞中IL-7 m RNA的表达水平,细胞计数法检测CAR-T细胞在GPC3抗原刺激下的增殖能力,ELISA检测CAR-T细胞在受到肿瘤细胞刺激后IL-7、IFN-γ和TNF-α的分泌水平。应用实时细胞分析(RTCA)技术检测CAR-T细胞对人肝癌Huh-7细胞的杀伤作用。结果:成功构建慢病毒载体GPC3-BBZ和GPC3-BBZ-NFAT-IL-7,制备出靶向GPC3的CAR-T细胞。经GPC3抗原激活后,GPC3-BBZ-NFAT-IL-7 CAR-T细胞可有效表达IL-7 mRNA(P<0.01),其表现出更强的增殖能力(P<0.05)。与GPC3-BBZ CAR-T细胞相比,GPC3-BBZ-NFAT-IL-7 CAR-T细胞与GPC3阳性靶细胞Huh-7细胞共培养后,分泌更高水平的IL-7、IFN-γ和TNF-α(P<0.01或P<0.001)。RTCA结果显示,GPC3-BBZ-NFAT-IL-7 CAR-T细胞对GPC3阳性Huh-7细胞的杀伤活性显著高于GPC3-BBZ CAR-T细胞(P<0.05)。结论:成功制备可诱导表达IL-7的靶向GPC3的CAR-T细胞,IL-7的诱导表达增强靶向GPC3 CAR-T细胞的免疫活性,在体外展现出较强的肿瘤细胞杀伤能力。

人参皂苷Rb1通过KEAP1/PGAM5/AIFM1通路促进肝细胞癌HepG2细胞发生氧死亡

目的:探讨人参皂苷Rb1(Gn-Rb1)对肝细胞癌(HCC)HepG2细胞氧死亡的影响及其可能的分子机制。方法:采用生物信息学方法分析氧死亡的关键基因PGAM5表达对HCC患者生存期的影响。选取辽宁省肿瘤医院收治的8例HCC患者的HCC组织与癌旁组织,通过WB法及qPCR检测氧死亡相关基因蛋白与mRNA的表达情况。将HepG2细胞随机分为对照组与Gn-Rb1组(予以200μmol/L Gn-Rb1干预),采用细胞克隆形成实验、划痕愈合实验分别检测Gn-Rb1对HepG2细胞的集落形成能力、迁移能力的影响,ELISA检测对细胞ROS生成水平的影响,微板法检测对细胞LDH释放水平的影响;WB法、qPCR法检测Gn-Rb1对HepG2氧死亡关键基因蛋白质与mRNA水平表达的影响。结果:生物信息学分析发现,PGAM5高表达肝癌患者总生存时间较低表达患者更长(P<0.05)。在临床HCC组织与癌旁组织样本中发现,相较于癌旁组织,在蛋白质与mRNA水平上,肿瘤组织KEAP1与PGAM5表达显著降低,NRF2表达显著升高(均P<0.01),p-AIFM1蛋白水平显著升高(P<0.05)。对HepG2细胞予以200μmol/L Gn-Rb1干预后,相较于对照组,Gn-Rb1组HepG2细胞的迁移能力与集落形成能力显著降低(均P<0.01),而LDH水平显著升高(P<0.05);相比于对照组,在mRNA和蛋白质水平上,Gn-Rb1组细胞中KEAP1、PGAM5表达均显著升高而NRF2表达均显著降低(均P<0.05),p-AIFM1蛋白表达显著降低(P<0.01)。结论:HCC组织中氧死亡被抑制,而Gn-Rb1能够通过调控KEAP1/PGAM5/AIFM1通路促进HepG2细胞氧死亡的发生,抑制细胞增殖和迁移能力。