Overexpression of anillin is related to poor prognosis in patients with hepatocellular carcinoma.

BACKGROUND: Anillin (ANLN) is required for tumor growth. It has been proven that knockdown of ANLN effectively reduces the occurrence of hepatocellular carcinoma (HCC) in transgenic mice. However, the functional role of ANLN in HCC patients remains to be elucidated. METHODS: Both microarray and TCGA project were used for the analyses of ANLN expression and regulation in HCC. The effect of ANLN on proliferation and cell cycle was detected by CCK-8, colony formation assay and flow cytometry. ANLN expression was measured by immunohistochemistry. Correlation between ANLN expression and clinicopathological features was assessed by Pearson Chi-square test and 5-year overall survival after liver resection was evaluated by Kaplan-Meier method. RESULTS: Increased copy number, decreased methylation levels in the CpG island and upregulated histone hypermethylation of ANLN were found in HCC. Knockdown of ANLN inhibited proliferation and induced G2/M phase arrest in SMMC-7721 cells. ANLN was mainly expressed in the nucleus and showed significantly higher expression levels in cancerous tissues than those in paired adjacent tissues. Moreover, nuclear ANLN expression levels in HCC metastases were significantly higher than those in primary HCC. The results of Cox proportional hazards regression model suggested that ANLN nuclear expression in HCC was an independent risk factor for poor 5-year overall survival of patients after liver resection. CONCLUSIONS: ANLN is a potential therapeutic target for HCC. Patients with nuclear ANLN overexpression in HCC tissue may need adjuvant therapy after liver resection.

Postoperative Complications and Survival Analysis of Surgical Resection for Hilar Cholangiocarcinoma: A Retrospective Study of Fifty-Nine Consecutive Patients.

Objective Hilar cholangiocarcinoma (HC) is invariably fatal without surgical resection. The primary aim of the current study was to determine the safety of variable surgical resections for patient with HC and their survival after surgical resection. In addition, prognostic factor for the overall survival was also evaluated. Methods The study included 59 consecutive patients who were newly diagnosed with HC and underwent surgical resections with curative intend between February 2009 and February 2017. Patients were followed up at 3-6 months intervals after hospital discharge. Postoperative complications and overall survival were determined. Associations of clinicopathologic and surgeon-related factors with overall survival were evaluated through univariate analysis and Cox regression analysis. Results Of patients with Bismuth and Corlette (B & C) type Ⅲ (n=19) and Ⅳ (n=25) HC lesions, 33 (55.9%) were treated with hilar resection combined with major liver resection (MLR), while the other 11 patients with type Ⅲ and Ⅳ, and those with type Ⅰ (n=8) and Ⅱ (n=7) HC lesions were treated with hilar resection. The overall surgical mortality was 5.1% and surgical morbidity was 35.6%. There was no statistical difference in the mortality between MLR group and hilar resection group (6.1% vs. 3.8%; X2=0.703, P=0.145). The median follow-up period was 18 months (range, 1-94 months). The 1-, 3-, 5-year survival rate was 59.3%, 36.5%, and 17.7%, respectively. The overall survival after resections was 18 months. In HC patients with B & C type Ⅲ and Ⅳ lesions, the median survival was 23 months for hilar resection with MLR and 8 months for hilar resection alone; the 1-, 3-, 5-year cumulative survival rate was 63.9%, 23.3%, and 15.5%, respectively for hilar resection with MLR, and 11.1%, 0, and 0, respectively for hilar resection alone, with significant differene observed (HR, 9.902; 95% CI, 2.636-19.571, P=0.001). Four factors were independently associated with overall survival: preoperative serum Ca19-9 (HR, 7.039; 95% CI, 2.803-17.678, P<0.001), histopathologic grade (HR, 4.964; 95% CI, 1.046-23.552, P=0.044), surgical margins (P=0.031), and AJCC staging (P=0.015). Conclusions R0 resection is efficacious in surgical treatment of HC. MLR in combination with caudate lobe resection may increase the chance of R0 resection and improve survival of HC patients with B & C type Ⅲ and Ⅳ lesions. Preoperatively prepared for biliary drainage may ensure the safety of MLR in most HC patients. Novel adjuvant therapies are needed to improve the survival of HC patients with poor prognostic factors.

Prognostic value of glypican-3 in patients with HBV-associated hepatocellular carcinoma after liver transplantation.

BACKGROUND: Glypican-3 (GPC-3) is frequently overexpressed in hepatocellular carcinoma (HCC). Recent studies have shown that GPC-3 is a highly efficient diagnostic biomarker of HCC and an indicator of poor prognosis in HCC patients who have undergone hepatectomy. However, its prognostic value in patients with HBV-associated HCC after liver transplantation (LT) is not clear. The present study is to evaluate the prognostic value of GPC-3 in patients with HBV-associated HCC after LT. METHODS: A cohort of 104 HCC patients with HBV-associated cirrhosis who had undergone LT at our hospital between 2002 and 2011 were enrolled in this study. Samples of HCC were taken from these patients. GPC-3 protein expression was detected in paraffin-embedded specimens using immunohistochemistry. All related clinical data were obtained from the China Liver Transplant Registry. The relationship between GPC-3 expression and clinicopathological parameters was analyzed. Univariate and multivariate Cox-regression analyses were used to identify risk factors for poor prognosis. RESULTS: GPC-3 was expressed in samples from 74 (71.2%) of the 104 patients. GPC-3 was expressed only in HCC cells. Positive staining was correlated with tumor size (P=0.004), encapsulation (P=0.018), pathological stage (P=0.027), portal vein invasion (P=0.043), tumor differentiation (P=0.002) and the Milan criteria (P=0.016). The 5-year survival rate and disease-free survival rate of patients with GPC-3-positive were lower than those (38.2% vs 75.4%, P<0.001; 30.8% vs 69.7%, P=0.001) of patients with GPC-3-negative. Multivariate Cox-regression analysis revealed that GPC-3 was an independent risk factor for 5-year survival rate (P=0.031) and disease-free survival rate (P=0.047), together with tumor differentiation, Milan criteria and pre-operative alpha-fetoprotein. CONCLUSION: GPC-3 is a potential biomarker for poor prognosis after LT in HCC patients with HBV-associated cirrhosis.

Esophagogastric devascularization without splenectomy in portal hypertension: safe and effective?

BACKGROUND: Esophagogastric variceal hemorrhage is a life-threatening complication of portal hypertension. In this study, we compared the therapeutic effect of a novel surgical procedure, esophagogastric devascularization without splenectomy (EDWS), with the widely used modified esophagogastric devascularization (MED) with splenectomy for the treatment of portal hypertension. METHODS: Fifty-five patients with portal hypertension were included in this retrospective study. Among them, 27 patients underwent EDWS, and the other 28 patients underwent MED. Patients' characteristics, perioperative parameters and long-term follow-up were analyzed. RESULTS: The portal venous pressure was decreased by 20% postoperatively in both groups. The morbidity rate of portal venous system thrombosis in the EDWS group was significantly lower than that in the MED group (P=0.032). The 1- and 3-year recurrence rates of esophagogastric variceal hemorrhage were 0% and 4.5% in the EDWS group, and 0% and 8.7% in the MED group, respectively (P=0.631). CONCLUSIONS: EDWS is a safe and effective treatment for esophagogastric varices secondary to portal hypertension in selected patients. Patients treated with EDWS had a lower complication rate of portal venous system thrombosis compared with those treated with conventional MED.

Hepatocellular carcinoma recurrence is an independent risk factor for HB recurrence after liver transplantation.

BACKGROUND/AIMS: To confirm the relationship between hepatitis B recurrence and Hepatocellular carcinoma recurrence. METHODOLOGY: Data from 340 patients undergoing liver transplantation for HBV-related liver disease were retrospectively evaluated. Clinically relevant variables were analyzed using univariate models. Significant variables were subjected to multivariate logistic regression analysis to identify the independent predictors for HBV recurrence. Fifteen samples removed from HCC recurrence patients were stained for HBsAg and HBcAg. RESULTS: The analyzed population included 283 male and 57 female patients. The mean age was 48.5±9.33 years and median follow-up was 47 months. Hepatitis B relapsed in 16 patients (4.7%). Univariate analysis indicated that HCC (P=0.022) and HCC recurrence (P=0.000) were associated to post transplantation HB. Multivariate analysis identified HCC recurrence as an independent risk factor for HB recurrence (hazard ratio: 23.262 (95% CI: 3.752, 144.216); P <0.001). Three of 15 metastatic lesions were positive for HBsAg and 1 lesion was positive for HBcAg. Conclusion: HCC recurrence is an independent risk factor for post transplantation recurrence of hepatitis.

Arachidonic acid activates NLRP3 inflammasome in MDSCs via FATP2 to promote post-transplant tumour recurrence in steatotic liver grafts.

Background & Aims: The steatotic grafts have been applied in liver transplantation frequently owing to the high incidence of non-alcoholic fatty liver disease. However, fatty livers are vulnerable to graft injury. Myeloid-derived suppressor cell (MDSC) recruitment during liver graft injury promotes tumour recurrence. Lipid metabolism exerts the immunological influence on MDSCs in tumour progression. Here, we aimed to explore the role and mechanism of inflammasome activation in MDSCs induced by lipid metabolism during fatty liver graft injury and the subsequent effects on tumour recurrence. Methods: MDSC populations and nucleotide-binding oligomerisation domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome levels were investigated in a clinical cohort and a rat liver transplantation model. The mechanism of NLRP3 activation by specific fatty acids was explored in mouse hepatic ischaemia/reperfusion injury (IRI) with tumour recurrence model and in vitro studies. Results: MDSC populations and NLRP3 levels were increased with higher tumour recurrent rate in patients using steatotic grafts. NLRP3 was upregulated in MDSCs with lipid accumulation post mouse fatty liver IRI. Mechanistically, arachidonic acid was discovered to activate NLRP3 inflammasome in MDSCs through fatty acid transport protein 2 (FATP2), which was identified by screening lipid uptake receptors. The mitochondrial dysfunction with enhanced reactive oxygen species bridged arachidonic acid uptake and NLRP3 activation in MDSCs, which subsequently stimulated CD4+ T cells producing more IL-17 in fatty liver IRI. Blockade of FATP2 inhibited NLRP3 activation in MDSCs, IL-17 production in CD4+ T cells, and the tumour recurrence post fatty liver IRI. Conclusions: During fatty liver graft injury, arachidonic acid activated NLRP3 inflammasome in MDSCs through FATP2, which subsequently stimulated CD4+ T cells producing IL-17 to promote tumour recurrence post transplantation. Impact and implications: The high incidence of non-alcoholic fatty liver disease resulted in the frequent application of steatotic donors in liver transplantation. Our data showed that the patients who underwent liver transplantation using fatty grafts experienced higher tumour recurrence. We found that arachidonic acid activated NLRP3 inflammasome in MDSCs through FATP2 during fatty liver graft injury, which led to more IL-17 secretion of CD4+ T cells and promoted tumour recurrence post transplantation. The inflammasome activation by aberrant fatty acid metabolism in MDSCs bridged the acute-phase fatty liver graft injury and liver tumour recurrence.

International experts consensus guidelines on robotic liver resection in 2023.

The robotic liver resection (RLR) has been increasingly applied in recent years and its benefits shown in some aspects owing to the technical advancement of robotic surgical system, however, controversies still exist. Based on the foundation of the previous consensus statement, this new consensus document aimed to update clinical recommendations and provide guidance to improve the outcomes of RLR clinical practice. The guideline steering group and guideline expert group were formed by 29 international experts of liver surgery and evidence-based medicine (EBM). Relevant literature was reviewed and analyzed by the evidence evaluation group. According to the WHO Handbook for Guideline Development, the Guidance Principles of Development and Amendment of the Guidelines for Clinical Diagnosis and Treatment in China 2022, a total of 14 recommendations were generated. Among them were 8 recommendations formulated by the GRADE method, and the remaining 6 recommendations were formulated based on literature review and experts' opinion due to insufficient EBM results. This international experts consensus guideline offered guidance for the safe and effective clinical practice and the research direction of RLR in future.

[Results of long-time follow up of patients who survived more than 5 years after liver transplantation: a single center experience].

OBJECTIVE: To investigate liver transplantation patients who survived for more than 5 years for the occurrences of their various long-term complications, prevention and treatment. METHODS: By May 31, 2010, totally 69 patients who had received liver transplantation from July 2000 to May 2005 in Peking University People's Hospital were still alive. We reviewed the clinical data of these patients and the recent records of their liver and kidney functions, blood pressure, blood sugar and blood fat, etc. The occurrences of their various long-term complications were summarized and the status of treatment was studied. RESULTS: In these 69 patients, 39.1% (27/69) of them were overweight or obese, 33.3% (23/69) had post transplantation diabetes mellitus (PTDM), 26.1% (18/69) had hyperlipemia, 20.3% (14/69) suffered from renal insufficiency, 15.9% (11/69) had hypertension and 23.2% (16/69) had hyperuricemia. Interestingly, the occurrences of PTDM and hyperlipemia in overweight or obese patients were higher than those in normal weight patients (48.2% vs. 23.8% and 40.7% vs. 16.7%, P<0.05). In addition, hepatitis B virus (HBV) infection recurred in 4 patients out of the 61 patients who had HBV related liver disease pre-operation, and liver cancer relapsed in 3 patients out of the 19 patients who had hepatocellular carcinoma (HCC) pre-operation. Totally 4 patients received re-transplant during the follow-up. CONCLUSION: The occurrences of long-term complications in liver transplantation patients who survived for more than 5 years were rather high, so the follow-up should be strengthened and procedures done to avoid the exacerbation of these complications.

Up-Regulation of Donor Dendritic Cell PD-L1 Expression Reduced Recipient Lymphocyte Activation and Proliferation In Vitro.

OBJECTIVE: To observe the effects of dendritic cells (DC) in donor C57BL/6 (H-2b) micetransfected with recombinant adenovirus vector Ad-PD-L1 on proliferation and activation of lymphocytes in recipient DBA/2 (H-2d) mice. METHODS: The pSport 1-mSD274 plasmid containing the full-length PD-L1 cDNA of the mouse was digested and subcloned to the shuttle plasmid pShuttle-GFP-CMV(-), and then the adenovirus skeleton plasmid pAdxsi-GFP-CMV-PD-L1 was constructed by enzymolysis and ligation, transformed into DH5α sensitive bacteria, and screened for positive clones. After enzyme digestion, sequencing, and identification, 293 cells were transfected with liposome after linearization for packaging and amplification, and the virus was purified by cesium chloride density gradient centrifugation. DC of donor C57BL/6 mice were isolated, cultured, and divided into the following 3 groups: group A, adenovirus vector Ad-PD-L1 transfection group; group B, empty vector transfection group; and group C, control group. Western blot was used to detect the expression of PD-L1 in each group of cells after transfection. Isolate lymphocytes from recipient DBA/2 mice were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) and mixed with DC of donor C57BL/6 mice with lymphocytes of recipient DBA/2 mice. Flow cytometry was performed to observe the proliferation of lymphocytes. RESULTS: Digestion and sequencing confirmed that the recombinant adenovirus vector Ad-PD-L1 containing PD-L1 was successfully constructed. After transfection with DC of donor C57BL/6 mice, the expression of PD-L1 increased by 37% (P < .05), and the PD-L1 transfected DC and recipient DBA/2. Mouse lymphocytes were cocultured. Compared with the control group, the increased expression of PD-L1 significantly inhibited the proliferation and activation of lymphocytes. The lymphocyte proliferation of DBA/2 mice decreased by 41% (P < .01). CONCLUSION: The recombinant adenovirus vector Ad-PD-L1 containing the mouse PD-L1 gene was successfully constructed. After transfection with dendritic cells of donor C57BL/6 mice, PD-1/PD-L1 inhibited lymphocytes proliferation and activation of recipient DBA/2 mice through costimulatory pathway.

NKG2D Enhances Double-Negative T Cell Regulation of B Cells.

Numerous studies reported a small subpopulation of TCRαß+CD4-CD8- (double-negative) T cells that exert regulatory functions in the peripheral lymphocyte population. However, the origin of these double-negative T (DNT) cells is controversial. Some researchers reported that DNT cells originated from the thymus, and others argued that these cells are derived from peripheral immune induction. We report a possible mechanism for the induction of nonregulatory CD4+ T cells to become regulatory double-negative T (iDNT) cells in vitro. We found that immature bone marrow dendritic cells (CD86+MHC-II- DCs), rather than mature DCs (CD86+MHC-II+), induced high levels of iDNT cells. The addition of an anti-MHC-II antibody to the CD86+MHC-II+ DC group significantly increased induction. These iDNT cells promoted B cell apoptosis and inhibited B cell proliferation and plasma cell formation. A subgroup of iDNT cells expressed NKG2D. Compared to NKG2D- iDNT cells, NKG2D+ iDNT cells released more granzyme B to enhance B cell regulation. This enhancement may function via NKG2D ligands expressed on B cells following lipopolysaccharide stimulation. These results demonstrate that MHC-II impedes induction, and iDNT cells may be MHC independent. NKG2D expression on iDNT cells enhanced the regulatory function of these cells. Our findings elucidate one possible mechanism of the induction of peripheral immune tolerance and provide a potential treatment for chronic allograft rejection in the future.

[Management of hepatic artery related complications and its outcome during liver transplantation].

OBJECTIVE: To explore the experience of hepatic arterial reconstruction and management of its complications. METHODS: The clinical data of 570 consecutive orthotopic liver transplantation patients performed from May 2001 to May 2009 in Peking University People's Hospital were analyzed retrospectively in order to summarize the key factors of hepatic arterial reconstruction and the experience of management of its complications. RESULTS: Arterial complications developed in 18 (3.1%) of the 570 patients including 11 cases of hepatic artery thrombosis, 5 cases of hepatic artery stenosis and 2 cases of hepatic artery rupture. Of the 11 cases with early complication (within 4 weeks), 7 patients died, including 2 due to the rupture of hepatic artery and 5 due to acute liver failure and sepsis of hepatic artery thrombosis. Of the 7 cases with late complication, 4 patients died, including 1 due to graft failure and 3 due to ischemic-type biliary complications. CONCLUSION: Good quality of donor artery and proper choice of microsurgical anastomosis technique in hepatic artery reconstruction could significantly reduce the incidence of its complication. Early detection and diagnosis with active early interventional therapy can improve prognosis of the patients.

[Liver transplantation for the patients with end stage liver disease and portal vein thrombosis].

OBJECTIVE: To investigate the outcome of liver transplantation(LT) for end stage liver disease with portal vein thrombosis (PVT) in different processes. METHODS: Data from 308 patients who underwent LT from July 2004 to February 2008 were retrospectively assessed. The processes of varies grades of PVT during LT were analyzed and estimated for whether the outcome of LT was different between patients with or without PVT. RESULTS: There were 46 patients with PVT, including 11 of grade 1, 14 of grade 2, 18 of grade 3 and 3 of grade 4. LT performed in grade 1 and 2 PVT patients without special intervention. LT was performed in 16 patients with grade 3 PVT after simple thrombectomy or thrombus-extraction. The other 2 patients with grade 3 PVT received the donor superior mesenteric vein to act as a bridge between the donor portal vein and host superior mesenteric vein. Two cases with grade 4 PVT received a cavo-portal hemitransposition, and the other one anastomosis between graft portal vein and varicose coronary vein. The postoperative 1-year survival rates of patients without PVT and patients with PVT were 91.6%(240/262), 80.5%(211/262)vs 86.9%(40/46), 76.1%(35/46), respectively. The patients with PVT had a recurrence rate of 4.3%(2/46). CONCLUSION: Most patients suffering from end stage liver disease with PVT can be successfully treated by LT. However, the result of the patients with diffused PVT undergoing LT is relatively poor.

[Analysis of 565 cases of liver transplantation at a single transplantation center].

OBJECTIVE: To determine the preventative and curative strategies after liver transplantation by investigating the risk factors on prognosis. METHODS: The data of 565 consecutive patients who underwent orthotopic liver transplantation were retrospectively analyzed with the survival rate and complication morbidity. RESULTS: The follow-up time ranges from 3 to 104 months of all the 565 patients after liver transplantation. From January 2004 to January 2009, the patient survival rates were 91.2% after 1 month, 84.9% after 1 year, 69.2% after 3 years, and 66.1% after 5 years, while they were 87.8%, 73.2%, 60.2%, and 57.7% from May 2000 to December 2003. The patient survival rates were 83.3% after 1 year, 79.8% after 3 years, and 78.5% after 5 years in non-hepatocellular carcimoma (non-HCC) group, while they were 78.4%, 49.1%, and 45.1% in HCC group. In early stage after surgery, the morbidities of re-operation due to intra-abdominal hemorrage, vascular complication, severe infection, acute renal failure and primary graft dysfunction were 1.1%, 1.6%, 13.6%, 7.4%, and 1.2%, while in late stage, the morbidities of HCC recurrence, biliary complications, HBV recurrence, de novo malignancy, chronic graft dysfunction were 40.3%, 6.7%, 2.1%, 0.9%, 0.9%, and 1.1%, respectively. The HCC recurrent rates were 8.1% versus 62.5% in matching Milan criteria group or exceeding Milan criteria group and the median survival time was 19.6 months of all recurrent patients. CONCLUSION: Liver transplantation has been the effective treatment for end stage liver disease. Due to the shortage of graft,we prefer to do operations for the patients without HCC or the patients with HCC but matching Milan Criteria.

Genetic alterations and expression of PTEN and its relationship with cancer stem cell markers to investigate pathogenesis and to evaluate prognosis in hepatocellular carcinoma.

AIMS: To investigate molecular alteration and expression of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene in hepatocellular carcinoma (HCC), and to evaluate the correlation between PTEN and cancer stem cell (CSC) markers and the prognostic value of these markers. METHODS: We evaluated changes of PTEN and CSC markers (CD133, epithelial cell adhesion molecule (EpCAM) and CK19) in 183 resection specimens by immunohistochemistry (IHC) and detected PTEN and phosphoinositide-3-kinase catalytic-alpha (PIK3CA) gene by fluorescence in situ hybridisation (FISH) in some specimens. RESULTS: PTEN and CD133, EpCAM and CK19 in 183 resection specimens were studied by IHC, and PTEN and PIK3CA genes were detected by FISH. PTEN expression was reduced in 92 HCC tissues (50.3%). There were 16 HCCs with PTEN deletion (51.6%). Comparison between PTEN IHC and FISH showed that the analysis was highly concordant (54/59, 91.5%). There were 19 HCCs with PIK3CA amplification. Deletion of PTEN was positively correlated with amplification of PIK3CA. Positive expression of CD133, EpCAM and CK19 was correlated with steatosis, moderate to poor differentiation, and so on. Reduction of PTEN expression was negatively correlated with positive expression of CD133, EpCAM and CK19. Reduced expression of PTEN (p=0.028) was an independent predictor for HCC recurrence and overall survival in HCC. PTEN-/CD133+ group had shorter OS and RFS time. CONCLUSIONS: PTEN plays a key role in hepatocarcinogenesis and reduction of PTEN expression is related to increased expression of CD133, EpCAM and CK19, which is a useful tool to evaluate HCC prognosis and recurrence.

Stromal morphological changes and immunophenotypic features of precancerous lesions and hepatocellular carcinoma.

AIMS: To evaluate stromal histopathological features and immunostaining expression for differential diagnosis of low- and high-grade dysplastic nodules (HGDN) to early and progressed hepatocellular carcinomas (eHCC, pHCC). MATERIALS: We evaluated sinusoid capillarisation (SC), solitary artery (SA), ductular reaction (DR), stromal invasion and expression of six biomarkers (GPC3, HSP70, GS, CD34, CK19, EpCAM) in a series of 97 cases. RESULTS: Stromal morphological changes, including SC, DR and SA, exhibited significant differences in differential diagnosis. In one indicator, SC had the best sensitivity (90.00%) and accuracy (85.42%), and SA had the best specificity at 88.89 %. In combinations, SC +and SA +were favourable and optimal. The immunoreactivity of GPC3, HSP70 and GS increased significantly in line with the stepwise progression of hepatocarcinogenesis. CONCLUSIONS: Stromal histopathology features are useful for diagnosing HGDN, eHCC and small HCC. The immunostaining panel of GPC3, HSP70 and GS can also be supplementary.

Paradoxical effects of cellular senescence-inhibited gene involved in hepatocellular carcinoma migration and proliferation by ERK pathway and mesenchymal-like markers.

BACKGROUND: Cellular senescence-inhibited gene (CSIG) strongly prolongs the progression of replicative senescence. However, roles and mechanisms of CSIG in tumor progression have not been studied widely. METHODS: Roles of CSIG in migration and proliferation of SMMC7721 and Huh7 cells were analyzed by transwell or cell viability assays, respectively. Tumorigenicity assays were used to study whether CSIG knockdown could affect SMMC7721 proliferation in vivo. Next, Western blotting and RT-PCR were preformed to evaluate the effects of CSIG on P-ERK cascade and epithelial mesenchymal transformation markers. Then, the location and expression of CSIG protein was detected by immunofluorescence and Western blotting, respectively. Finally, the Cancer Genome Atlas dataset was used to analyze CSIG mRNA levels in hepatocellular carcinoma (HCC) and adjacent non-tumor tissues. RESULTS: In this study, we found that CSIG overexpression promoted SMMC7721 cell migration, and CSIG knockdown suppressed tumorigenicity of SMMC7721 cells. In contrast to expectation, CSIG up-regulation could significantly inhibit Huh7 cell growth and migration. CSIG could promote P-ERK activation and levels of mesenchymal-like markers in SMMC7721 cells, whereas CSIG suppressed P-ERK activation and levels of mesenchymal-like markers in Huh7 cells. CSIG protein was located in nucleoli as well as nucleoplasm of SMMC7721 cells, whereas CSIG protein was mainly expressed in the nucleoli rather than nucleoplasm of Huh7 cells. Finally, due to individual differences, raised or down-regulated trends of CSIG in HCC as compared with adjacent non-tumor tissues are different among various patient populations. CONCLUSION: In summary, these results indicate that CSIG might play different roles in SMMC7721 and Huh7 cells through regulating P-ERK pathway and mesenchymal-like markers. The differential distribution of CSIG might be an important factor that causes its different functions in SMMC7721 and Huh7 cells. CSIG might play different roles in various patient populations.

Transarterial chemoembolization versus best supportive care for patients with hepatocellular carcinoma with portal vein tumor thrombus：a multicenter study.

BACKGROUND: This study aims to compare the efficacy and safety of treatment after transarterial chemoembolization(TACE) with best supportive care (BSC) in patients with hepatocellular carcinoma (HCC) with PVTT. METHODS: This retrospective study was conducted on 1,040 patients with HCC with PVTT who were treated either with TACE (n = 675) or BSC (n = 365). BSC did not include sorafenib. The two groups of patients were compared with or without propensity score matching. A subgroup analysis was subsequently performed by stratifying patients according to the stages of PVTT in the Cheng's PVTT classification. RESULTS: In PVTTtypes I-III, TACE was associated with significantly better overall survival (OS) thanBSC (P < 0.05). Within each type of PVTT for patients who received TACE or BSC, OS was significantly worse in patients with type IVPVTT than in any of the other three types of PVTT (all P < 0.05). TACE was associated with better long-termOS than BSC after propensity score matching or on stratification by the PVTT types. CONCLUSION: TACE was associated with better OS than BSC in HCC patients with PVTT types I-III but not type IV. Patients with type IV PVTT showed the worst prognosis, regardless of whether TACE or BSC was used.