GCNT3 regulated MUC13 to promote the development of hepatocellular carcinoma through the GSK3ß/ß-catenin pathway.

BACKGROUND: Hepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths worldwide. Extensive research is currently directed at identifying novel targets for its diagnosis and treatment. AIMS: We investigated the biological functions and clinical significance of mucin-type N-acetylglucosaminyltransferase 3 (GCNT3) in HCC. METHODS: Variations in the mRNA expression of GCNT3 were examined in normal and HCC tissues. Cell function assays and animal models characterized the effects of GCNT3 on the proliferation, invasion, and migration abilities of HCC cells. Western blot and immunofluorescence analyses were performed to explore further the specific mechanisms whereby GCNT3 affects HCC progression. RESULTS: There is a strong correlation between GCNT3 overexpression and tumor formation and metastasis in vivo and in vitro. GCNT3 acted as a regulator of the synthesis of mucin-type O-glycans by interacting with mucin 13 (MUC13) to regulate its expression levels, activating the GSK3ß/ß-catenin signaling pathway. The activation of GSK3ß/ß-catenin signaling by GCNT3 was mitigated by MUC13 knockdown. In clinical HCC specimens, GCNT3 expression was upregulated in HCC tissues compared to non-tumor tissues. Further, there was a significant correlation between high GCNT3 expression and poor patient survival. CONCLUSIONS: GCNT3 regulated tumor progression in HCC through the MUC13/GSK3-ß/ß-catenin signaling pathway.

Effect of Microvascular Invasion on the Prognosis in Hepatocellular Carcinoma and Analysis of Related Risk Factors: A Two-Center Study.

Objective: Microvascular invasion is considered to initiate intrahepatic metastasis and postoperative recurrence of hepatocellular carcinoma (HCC). We aimed to analyze the effect of MVI on the prognosis in HCC and identify related risk factors for microvascular invasion (MVI). Methods: The clinical data of 553 HCC patients who underwent liver surgery at Qingdao University from January 2014 to December 2018 and 89 patients at Beijing Tsinghua Changgung Hospital treated between October 2014 and October 2019 were collected retrospectively. We explored the impact of MVI on the prognosis of patients with HCC using Kaplan-Meier analysis. We conducted logistic regression analysis to identify variables significantly related to MVI. Results: Pathological examination confirmed the presence of MVI in 265 patients (41.3%). Six factors independently correlated with MVI were incorporated into the multivariate logistic regression analysis: Edmondson-Steiner grade [odds ratio (OR) = 3.244, 95%CI: 2.243-4.692; p < 0.001], liver capsule invasion (OR = 1.755; 95%CI: 1.215-2.535; p = 0.003), bile duct tumor thrombi (OR = 20.926; 95%CI: 2.552-171.553; p = 0.005), α-fetoprotein (> 400 vs. < 400 ng/ml; OR = 1.530; 95%CI: 1.017-2.303; p = 0.041), tumor size (OR = 1.095; 95%CI: 1.027-1.166; p = 0.005), and neutrophil-lymphocyte ratio (OR = 1.086; 95%CI: 1.016-1.162; p = 0.015). The area under the receiver operating characteristic curve (AUC) was 0.743 (95%CI: 0.704-0.781; p < 0.001), indicating that our logistic regression model had significant clinical usefulness. Conclusions: We analyzed the effect of MVI on the prognosis in HCC and evaluated the risk factors for MVI, which could be helpful in making decisions regarding patients with a high risk of recurrence.

Clinicopathologic features of hepatocellular carcinoma patients surviving more than 10 years after radical hepatectomy / 中华外科杂志

Objective@#To clarify the clinicopathologic features of hepatocellular carcinoma (HCC) patients survived more than 10 years after radical hepatectomy.@*Methods@#Two hundreds and fifty-two patients who underwent curative resection for HCC between January 1999 and March 2006 at Department of Hepatopancreatobiliary Surgery, Affiliated Hospital of Qingdao University were included.There were 217 male cases and 35 female cases aging from 17 to 82 years with median age of (53.8±10.5)years. Followed by March 31 2016, clinicopathologic factors in 10-year survivors and patients who died within 10 years were compared by χ2 test, Kaplan-Meier survival analysis and Cox proportional hazards model and the prognostic factors affecting survival were identified.@*Results@#All patients were followed-up for 4.0 to 205.7 months with median time of 53.4 months. The 10-year overall survival rate was 26%, there were 62 cases(26.2%) who survived for more than 10 years after initial hepatectomy. In survival >10-year group, the paitents with ALT<40 U/L, gamma-glutamyl transpeptidase<64 U/L, albumin≥35 g/L, without liver cirrhosis and portal hypertension, Child-Pugh grade A, no blood transfusion, AFP≤20 μg/L, tumor size ≤5.0 cm, single tumor, high differentiation, TNM stage Ⅰ and TACE negative after resection were more than the patients in survival <10-year group (P<0.05). In multivariate analysis, Child-Pugh grade A, the tumor size ≤5.0 cm and TACE negative after resection were favorable independent factors associated with 10-year survival (P<0.05).@*Conclusion@#Based on the results of the study, Child-Pugh grade A, tumor size ≤5.0 cm and TACE negative after resection at initial hepatectomy might be biologically favorable conditions for patients surviving more than 10 years.

Expression of Slug in pancreatic cancer and inhibitory effects of anti-Slug on invasion and migration of pancreatic carcinoma cells / 中华肝胆外科杂志

Objective To investigate expression of slug and E-cadherin in pancreatic cancer tissues and determine the inhibitory effects of anti-Slug, an anti-sense plasmid, on the invasion of pancreatic cancer cell lines in vitro. Methods Slug and E-cadherin protein and mRNA was analyzed by IHP and RT-PCR in 36 cases of pancreatic cancer. Then anti-Slug plasmid was transfected into herin and Slug expression. The inhibitory effects of anti-sense Slug were also detected by Transwell motility assay and Matrigel invasion assay. Results The expression of Slug and mRNA in metastatic pancreatic cancer tissue was higher than that in non-metastatic tissue. E-cadherin and mRNA was lower in metastasis tissues(P＜0.05). The inverse relationships were further observed by transient transfection of anti-Slug into SW1990H4 cells. The downregulated expression of Slug and re-expression of E-cadherin were found. The Slug mRNA levels were 0.985±0.016,0.973±0.014, 0. 554±0. 011 after 0, 48 h of transfection of anti-sense Slug, and that of E-cadherin were 0.120±0.001, 0.360±0.002, 0. 727±0. 006, respectively. The diference was significant between different time points (P＜0.05). The Slug mRNA levels were 0. 206±0.017, 0.968±0.015, and that of E-cadherin were 0. 18±0.002,0.727±0.006 after stable transfection of anti-sense Slug, and control plasmid, respectively. The diference was significant (P＜0.05). The motility activity(393±28, 352±24, 96 ±13 )and the invasion activity (223 ± 69, 202 ± 64, 65 ±19) of1 antisense Slug transfectant cells were significantly decreased as compared with those of control cells (P＜0.05). Conclusions Higher expression of slug and lower expression of E-cadherin is related to the invasion and metastasis in pancreatic cancer. A reverse corelation of E-cadherin and Slug expression exists in pancreatic cancer. Slug is possibly a potential target for cancer gene therapy blocking invasion and metastasis in human pancreatic cancer.

Relationship between nuclear factor-κB as well as p53 up-regulated modulator of apoptosis and lung injury induced by severe acute pancreatitis and therapeutic effect of proline dithiocarbamate / 中华急诊医学杂志

Objective To investigate the expression of nuclear factor-κB (NF-κB) and p53 up-regulated modulator of apoptosis (PUMA) in acute lung injury (ALI) induced by severe acute pancreatitis (SAP), and the therapeutic role of proline dithiocarbamate (PDTC). Method SD rats weighed 200～ 250 g were randomly(random number) divided into sham operation group (A group, n = 18), ALI group (B group, n = 18) and PDTC treatment group (C group, n = 18). The model of SAP was eastablished by injecting 1 mL/kg of sodium tauarocholate into the pancreatic capsule of the rats in B group and C group. The model rats in C group were treated with PDTC one hour after modeling. Six rats of each group were sacrificed 6 h,12 h, and 24 hours after modeling. The histopathological changes in lung and pancreas were observed. The levels of NF-κB p65 and PUMA in lung were detected by using Western blotting, and the expressions of bcl-2, bax and caspase-3 mRNA in the lung were detected by using RT-PCR. The lung tissue was taken for examination under transmission electron microscope. TUNEL was used for detection of apoptotic alveolar epithelial cells. Results Six to 24 hours after modeling, the pathological scores in lung of ALI group were significantly higher than those of control group and PDTC group after sodium taurocholate injection ( P ＜ 0.05). The levels of NF-κB p65 and PUMA, and the expressions of bax and caspase3 mRNA in ALI group at different intervals were higher than those in control group and PDTC group ( P ＜ 0.05),whereas the expression of bcl-2 mRNA in ALI group was lower than that in control group and PDTC group ( P ＜0.05). The NF-κB p65 was correlated closely and positively with PUMA ( r= 0.987, P ＜ 0.01). Higher activity of caspase-3 acrtive units was seen in ALI group than that in control group and PDTC group ( P ＜ 0.05). The microvilli disappeared in ALI group 24 hours later. The apoptosis index in ALI group was higher than that in control group and PDTC group ( P ＜ 0.05). Conclusions The apoptosis of alveolar epithelial cells of rats in ALI group is caused by PUMA activated by NF-κB. PDTC treatment can inhibit apoptosis of alveolar epithelial cells of rats in ALI group by inhibiting the activation of NF-κB.

Radiosensitivity of AsPC-1 cell to γ-rays enhanced by up-regulation of PUMA induced by targeted Slug gene / 中华放射医学与防护杂志

Objective To explore the influence of PUMA on radiosensitivity of pancreatic cancer AsPC-1 cells after Slug gene inhibition by transfected short interferencing RNA(siRNA). Methods The AsPC-1 cells were infected with MOI 10,50,100 for 72 h, respectively. The expression of Slug and PUMA was analyzed by Western blotting and immunohistochemistry methods. The transfected and control cells were exposed to 4 Gy γ-rays. The cells inhibition rate was examined by MTT, Hoechst 33342 and IP double staining. DNA ladder and Giemsa staning was used to observe apoptosis. Results The relative value of Slug expression was 0.831 ±0.14,0. 546 ±0.12 and 0.178 ±0.08 after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly lower than that of control group ( F = 4. 992,P ＜ 0.05 ).The relative value of PUMA was 0. 325 ±0. 07,0. 593 ±0. 11 and 0. 978 ±0. 12, after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly higher than that of control group ( F = 4. 324,P ＜ 0. 05 ). The cell proliferation rate was ( 78.76 ± 9. 36 ) % in transfection combined with radiosensitivity group, significantly higher than that of transfection group [ ( 43.68 ± 6.71 ) % ] and radiosensitivity group alone [( 19.25 ± 3.72)% ] (F = 5.056, P ＜ 0.05). The apoptosis of transfection combined with radiosensitivity group was significantly higher than that of others. Conclusions Slug gene targeting siRNA could inhibit the expression of Slug, and consequently increase the activation of PUMA expression, and so enhance the radiosensitivity to γ-rays.