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Objective To establish a nomogram for overall survival rate after liver resection for primary small hepatocellular carcinoma based on SEER data and external validation of Chinese data. Methods The data of 1809 patients, registered in National Cancer Institute SEER database in 2004-2015, who underwent hepatectomy for primary small hepatocellular carcinoma were extracted as modeling group, and 158 patients with small hepatocellular carcinoma who underwent hepatectomy in Affiliated Hospital of North Sichuan Medical College from 2010 to 2017 were collected as validation group. The univariate Cox risk regression analysis, lasso regression analysis, and multivariate Cox hazard regression analysis were used to investigate the influencing factors for OS after hepatectomy in patients with small hepatocellular carcinoma. A nomogram was established based on the independent influencing factors for OS, and index of concordance (C-index), calibration curves, and receiver operating characteristic (ROC) curve were used to analyze the predictive ability of the nomogram. The Kaplan-Meier survival analysis and the log-rank test were used to investigate the difference in survival between the high- and low-risk groups. Results The multivariate Cox hazard regression analysis showed that sex (hazard ratio [ HR ]=1.22, 95% confidence interval [ CI ]: 1.05-1.41, P =0.010), Seer stage ( HR =1.51, 95% CI : 1.23-1.85, P < 0.001; HR =10.31, 95% CI : 2.53-42.04, P =0.001), tumor diameter ( HR =1.22, 95% CI : 1.06-1.39, P =0.004), vascular invasion or metastasis ( HR =1.43, 95% CI : 1.24-1.65, P < 0.001), and alpha-fetoprotein ( HR =1.33, 95% CI : 1.16-1.54, P < 0.001) were independent risk factors for OS after hepatectomy for small hepatocellular carcinoma. The modeling group had a C-index of 0.621, and its area under the ROC curve at 1, 2, and 3 years was 0.666(95% CI 0.628-0.704), 0.678(95% CI 0.647-0.708), and 0.663(95% CI : 0.635-0.690), respectively; the validation group had a C-index of 0.718, and its area under the ROC curve at 1, 2, and 3 years was 0.695(95% CI : 0.593-0.797), 0.781(95% CI : 0.706-0.856), and 0.759(95% CI 0.669-0.848), respectively. Risk stratification was performed based on the nomogram, and the Kaplan-Meier survival analysis showed that for both the modeling group and the validation group, the low-risk group had a significantly better prognosis than the high-risk group ( P < 0.01). Conclusion The model established for survival rate after liver resection for primary small hepatocellular carcinoma can predict the 1-, 2-, and 3-year OS rates and can thus be used in clinical practice in China.
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Objective To establish a nude mouse model of low rectal cancer with lymph node metastasis, to detect the expression of E-calcium (E-cad) in the serum and transplanted tumor tissue of the mice, and to analyze the role of E-cad in lymph node metastasis of rectal cancer.Methods Human colorectal adenocarcinoma cells (SW-480) were inoculated into the rectal submucosa of nude mice to establish a model of low rectal cancer with lymph node metastasis.120 nude mice were randomly divided into three groups (40 mice in each group).Group A (rectal cancer model group) was injected with dimethylhydrazine and implanted with SW480 cells.Group B (dimethylhydrazine control group) was injected with dimethylhydrazine alone.The group C (control group) received no treatment.Serum E-cad was detected by ELISA.The mRNA and protein expression of E-cad in the tumor tissue was detected by RT-PCR and western blot, respectively.Results The serum level of E-cad in group A was 19.48±1.25 mg/L, significantly higher than those in groups B (2.36±0.18 mg/L) and C (2.15±0.12 mg/L) (t =8.28, 9.01, P < 0.05).The serum levels of E-cad were significantly higher in the nude mice with lateral lymph node metastasis than those without lymph node metastasis (t =10.28, P < 0.05).The protein expression of E-cad in the group A was lower than that in the groups B and C (t =9.81, 7.69, P < 0.05).In the group A, the protein expression of E-cad in the nude mice with lateral lymph node metastasis was significantly lower than that without lateral lymph node metastasis (t =9.36, P < 0.05).The mRNA expression of E-cad in the group A was significantly lower than that in the groups B and C (P < 0.05), and the mRNA expression of E-cad in the nude mice with lateral lymph node metastasis was significantly lower than that in the mice without lateral lymph node metastasis (t =7.85, P < 0.05).Conclusions The serum level of E-cad may be closely associated with the lymphatic metastasis of rectal cancer, and the mRNA and protein expressions of E-cad in colorectal cancer tissue were weak or absent, leading to a decrease of adhesion ability between cancer cells, and promote the invasion and metastasis of cancer cells.
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Objective To explore the effect and mechanism of Gli2 on EMT and invasion in the hepatocellular car-cinoma cell line SMMC-7721.Methods shRNA of Gli2 and Negative control (NC) shRNA were constructed and transfected into SMMC-7721 cells.shRNA-Gli2 group,shRNA-NC group and blank group were set up .Transwell chambers assay , adhesion experiments were used to detect the ability of invasion ,homogeneous and heterogeneous cells intercellular adhesion of each group .Meanwhile, the qRT-PCR, Western blot were used to examine Gli2, E-cadherin ,N-cadherin and vimentin mRNA and protein expression .Results In different hepatocellular carcinoma cell lines and be along with the increasing ability of the invasion in hepatocellular carcinoma cell lines , Gli2 expres-sion was higher .Compared with the shRNA-NC group and blank control group ,the interfered group extensive cells invasion ability was inhibited ( P nism may be related to the up-regulation of E-cadherin and the dow N-regulation of N-cadherin, vimentin.
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Objective To detect the expression of ARK5 in hepatocellular carcinoma (HCC)tissue and hepatoma SMMC-7721 cells,and to investigate its effect on the growth of hepatoma cells.Methods The expression levels of ARK5 mRNA and protein were determined by RT-PCR and Western blotting in 30 cases of HCC tissue, paracarcinoma tissue,SMMC-7721 cells,and hepatic cells LO2.The SiRNA of ARK5 and negative control (NC) siRNA were constructed and transfected into the SMMC-7721 cells,and used as experimental group and negative control group;at the same time blank control group was set up. The proliferation activity and apoptotic rate of transfected cells were detected by MTT assay and flow cytometry (FCM).Results The PCR and Western blotting results showed that the expression levels of ARK5 mRNA and protein in HCC tissue and SMMC-7721 cells were significantly higher than those in paracarcinoma tissue and LO2 cells (P<0.05 ). The MTT assay results demonstrated that the inhibitory rates of growth of transfected cells in experimental group at 24,48 and 72 h were (19.39±5.42)%, (23.19±0.53)%,and (20.74±1.23)%;there were significant differences compared with blank control group and negative control group (P<0.01).The FCM results indicated that the apoptotic rate of the transfected cells in experimental group was (15.017±0.945)%,there were significant differences compared with blank control group (8.770%± 0.656 )% and negative control group (8.763%± 1.201%) (P<0.05 ). Conclusion The ARK5 expression level is significantly increased in HCC tissue and hepatoma SMMC-7721 cells;the inhibition of ARK5 expression could suppress the growth of hepatoma cells and induce apoptosis. So ARK5 maybe act as a cancer-promoting gene and induce hepatocellular carcinogenesis.
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Objective:To explore the effect of calcium release-activated calcium channel modulator 1 (ORAI1) on the migration and invasion of colon cancer cell line SW480 and its mechanism. Methods:The SW480 cells were infected with ORAI interference lentivirus. The expression of ORAI1 mRNA and protein was confirmed by quantitative real-time polymerase chain reaction and Western blot. Transwell chamber, adhesion, angiogenesis, and vasculogenic mimicry experiments were conducted to detect the ability of cell invasion, migration, and angiogenesis and the intercellular adhesion of homogeneous and heterogeneous cells among each group. Confocal microscopy was employed to detect the difference of store-operated Ca2+entry (SOCE) in each group. Western blott was used to detect the expression of ERK1/2, p-ERK1/2, MMP-2, VEGF, and E-cadherin protein. Results:After the infection of SW480 with the ORAI1 interference lentivirus for 72 h, significant fluorescence expression was observed. Compared with the empty vector group and control group, the expression of ORAI1 was lower in the interference group (P<0.01). Invasion and migration ability decreased (P<0.01); the intercellular adhesion ability of homogeneous cells increased (P<0.05); the intercellular adhesion ability of heterogeneous cells decreased (P<0.05);the angiogenesi and vasculogenic mimicry were enhanced (P<0.01);the internal flow peak of SOCE was low (P<0.05); the expression of p-ERK1/2, MMP-2, and VEGF proteins decreased (P<0.01); and the expression of E-cadherin protein increased (P<0.01). Conclusion:ORAI1 may promote the migration and invasion of SW480. This mechanism may be associated with the increase of SOCE.