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Objective To investigate the expression and the potential roles of long non-coding RNA(lncRNA)cancer susceptibility candidate 2(CASC2)and imprinted gene H19 in extrahepatic cholangiocarcinoma(ECC). Methods Four samples from patients with ECC were collected for high-throughput sequencing which was conducted to reveal the transcriptomic profiles of lncRNA CASC2 and H19.Bioinformatics tools were employed to predict the potential roles of the two genes.Another 22 ECC tissue samples and the cholangiocarcinoma cell lines(RBE,QBC939,HuH-28,and HuCCT1)with different degrees of differentiation were selected for validation.The para-carcinoma tissue and normal human intrahepatic biliary epithelial cell(HIBEC)were used as the control groups.The expression levels of lncRNA CASC2 and H19 in carcinoma tissue,para-carcinoma tissue,and cell lines were determined by real-time quantitative polymerase chain reaction(qRT-PCR).The correlation analysis was carried out for the clinical indicators of patients with the expression levels of the target genes. Results The two target genes showed significantly different expression between carcinoma tissue and para-carcinoma tissue(all P<0.05).Specifically,CASC2 had higher expression level in the carcinoma tissue than in the para-carcinoma tissue(t=1.262,P=0.025),whereas the expression of H19 showed an opposite trend(t=1.285,P=0.005).The expression levels of CASC2 in QBC939(t=8.114,P=0.015)and HuH-28(t=9.202,P=0.012)cells were significantly higher than that in the control group.The expression levels of H19 were significantly lower in RBE(t=-10.244,P<0.001),QBC939(t=-10.476,P<0.001),HuH-28(t=-19.798,P<0.001),and HuCCT1(t=-16.193,P=0.004)cells than in the control group.Bioinformatics analysis showed that CASC2 was mainly involved in the metabolic process and H19 in the development of multicellular organisms.Both CASC2 and H19 were related to catalytic activity.The expression level of lncRNA CASC2 was correlated with pathological differentiation(χ 2=6.222,P=0.022)and lymph node metastasis(χ2=5.455,P=0.020),and that of lncRNA H19 with pathological differentiation(χ2=1.174,P=0.029)and tumor size(χ2=-0.507,P=0.037). Conclusions In the case of ECC,lncRNA CASC2 and H19 have transcription disorders.lncRNA CASC2 is generally up-regulated in the carcinoma tissue,while H19 is down-regulated.Both genes have the potential to become new molecular markers for ECC.
Assuntos
Humanos , Neoplasias dos Ductos Biliares/genética , Ductos Biliares Intra-Hepáticos/metabolismo , Colangiocarcinoma/genética , Regulação Neoplásica da Expressão Gênica , RNA Longo não Codificante/genética , Proteínas Supressoras de Tumor/genéticaRESUMO
Objective To investigate the correlations between expression of CASC2 and hepatocellular carcinoma(HCC) prognosis.Methods A total of 129 patients including 80 males and 49 females with HCC were includedin this study,ranging from 21 to 73 years in Xuanwu Hospital of Capital Medical University and Beijing You'an Hospital were retrospectively analyzed from September 2007 to January 2014.Expression of CASC2 was assessed using reverse transcription quantitative-polymerase chain reaction in HCC tissue and the adjacent normal tissue.The correlations between CASC2 mRNA level and clinicopathological parameters was investigated.The relationship between the expression of CASC2 and the prognosis of patients with HCC was analyzed by Kaplan-Meier method.A log-rank analysis was performed to identify group differences.Univariate and multivariate Cox analysis were used to analyze the variables affecting the patient's prognosis.Results In 129 HCC samples,the level of CASC2 expression (0.84 ± 0.05) was lower than (3.35 ± 0.11) adjacent normal tissue (P ＜ 0.05).There were significant differences between CASC2 expression and tumor size,histological differentiation,and tumor stage in 129 HCC speciments.The median expression level of CACS2 in HCC tissues,0.84-fold,was used as the cut-off value to divide the 129 patients into two groups:low-expression group (n =72) and high-expression group (n =57).Overall survival rate of HCC patients with high CACS2 expression was significantly higher than those of patients with low CACS2 expression(P ＜0.05).Multivariate analysis indicated that histological differentiation (HR =0.20,95％ CI:0.05 ～ 0.59),tumor stage (HR =1.71,95％ CI:1.02 ～ 2.99) and CACS2 expression (HR =O.51,95％ CI:O.08 ～0.92) were an independent predictor of overall survival.Conclusion Low expression of CACS2 might be associated with the occurrence and development of HCC.
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Objective There are few reports on the relationship between lncRNA cancer susceptibility candidate gene 2 (CASC2) and NF-κB signaling pathway in thyroid papillary carcinoma cells at home and abroad. This study aimed to investigate the effect of lncRNA CASC2a on the proliferation and migration of TPC-1 via NF-κB signaling pathway. Methods TPC-1 was selected and constructed into lncRNA CASC2a overexpression plasmids which were divided into plasmid group [transfection with overexpressed plasmid pcDNA3.1(+)-CASC2a], empty group [transfection with equal amount of pcDNA3.1 (+) empty plasmid], blank group (without any processing). The effect of overexpressed lncRNA CASC2a on the expression of CASC2a in each group of TPC-1 cells was examined. The cells of transfected plasmid group were randomly divided into transfection group [transfection with only overexpressed plasmid pcDNA3.1(+)-CASC2a], transfection + PMA group [transfection with overexpressed plasmid pcDNA3.1(+)- CASC2a + propylene glycol methyl ether acetate (PMA) stimulation], control group (without any processing), PMA group (only plus PMA stimulation). The effects of lncRNA CASC2a on cell proliferation and migration were verified by CCK-8 and cell scratch assays at 24, 48, 72 and 96 h. Western blot was used to detect the expression of p105/p50 and p65 in NF-κB signaling pathway. The expression of NF-κB signaling pathway-related antibody proteins p105/p50 and p65 was observed by NF-κB signaling pathway agonist PMA(propylene glycol methyl ether acetate). Results After transfection with overexpressed plasmid pcDNA3.1(+)-CASC2a, the expression of CASC2a in plasmid group was significantly higher than that in empty group (P<0.05). The cell proliferation ability (0.191±0.005) was significantly lower in transfection+PMA group than that in transfection group (0.217±0.013), PMA group (0.247±0.009), and control group (0.260±0.004), and the difference was statistically significant ( P<0.01), while the cell proliferation ability in transfection group was significantly lower than those in PMA group and control group (P<0.01). The cell migration ability of transfected + PMA group [(0.208±0.109)%] was lower than that of transfection group, PMA group, and control group [(1.775±0.061)%, (1.622±0.519)%, (2.927±0.136)%], while the cell migration ability of transfection group and PMA group was lower than that in control group (P<0.05). The relative expression of p105/p50 and p65 protein in plasmid group was significantly lower than that in blank group (P<0.05). The expression of p105/p50 and p65 protein in transfection+PMA group [(0.674±0.007), (0.713±0.014)] was significantly higher than that in transfection group [(0.581±0.003), (0.570±0.012)] (P<0.05). Conclusion lncRNA CASC2a may inhibit the proliferation and migration of thyroid papillary carcinoma cells through NF-κB signaling pathway.