长链非编码RNA胃癌相关转录物3在肝癌中的表达及其对HepG2细胞生物学特性的影响

Hongwei Xu, Yutian Luo, Dajun Wang, Liang Wang
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Methyl thiazol tetrazolium (MTT) assay was used to detect cell proliferation in each group at 24, 48 and 72 h. The cell migration and invasion were detected by cell scratches experiments and Transwell assay, and the expression of Wnt/β-catenin signaling pathway protein was detected by Western blotting. \n \n \nResults \nThe expression of lncRNA GACAT3 in hepatocellular carcinoma tissues (3.10±1.93) was significantly higher than that in adjacent tissues (1.02±0.59, t=8.086, P<0.01); The expression of lncRNA GACAT3 in PLC/PRF/5 (1.92±0.21, t=7.074, P<0.01), Hep3B (2.42±0.25, t=9.327, P<0.01) and HepG2 cells(2.96±0.31, t=10.582, P<0.01) were significantly higher than that in normal liver cells THLE-3 (0.97±0.10). Compared with the silencing control group (1.03±0.11), the expression of lncRNA GACAT3 in the GACAT3 group (0.16±0.02) was significantly decreased (t=13.478, P<0.01). Cell proliferation was significantly reduced at 48 h and 72 h (P<0.01). Scratch healing rate and migratory and invasive ability were significantly decreased (P<0.01). The expression of Wnt/β-catenin signaling pathway protein β-catenin (0.15±0.02) and its downstream proteins c-Myc (0.22±0.02) and Cyclin D1 (0.21±0.02) were significantly lower than that of the silencing control group(0.81±0.08, t=13.863; 0.65±0.07, t=9.731; 0.78±0.08, t=10.230; P all<0.01). The expression of p-β-catenin protein (0.75±0.08) was significantly higher than that of the silencing control group (0.27±0.03, t=11.972, P<0.01). \n \n \nConclusion \nLncRNA GACAT3 is highly expressed in hepatocellular carcinoma. 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引用次数: 0

摘要

目的探讨长链非编码RNA(lncRNA)胃癌相关转录物3(GACAT3)在肝细胞癌中的表达及其对HepG2细胞增殖、侵袭和迁移的影响及其可能的机制。方法应用实时定量聚合酶链反应(real-time PCR)检测lncRNA GACAT3在癌症组织、癌旁组织、PLC/PRF/5、Hep3B、HepG2细胞系和正常肝细胞THLE-3中的差异表达。将阴性对照小干扰RNA(siRNA)和lncRNA GACAT3 siRNA转染到HepG2细胞中,48小时后,用实时PCR检测转染细胞中lncRNA GAPAT3的表达。采用甲基噻唑四唑蓝(MTT)法检测各组在24、48和72小时的细胞增殖。通过细胞划痕实验和Transwell法检测细胞迁移和侵袭,并通过Western印迹检测Wnt/β-catenin信号通路蛋白的表达。结果肝细胞癌组织中lncRNA GACAT3的表达(3.10±1.93)明显高于癌旁组织(1.02±0.59,t=8.086,P<0.01);lncRNA GACAT3在PLC/PRF/5(1.92±0.21,t=7.074,P<0.01)、Hep3B(2.42±0.25,t=9.327,P<0.01)和HepG2细胞(2.96±0.31,t=10.582,P<0.01)中的表达显著高于正常肝细胞THLE-3(0.97±0.10),GACAT3组lncRNA GACAT3的表达(0.16±0.02)显著降低(t=13.478,P<0.01),细胞增殖在48小时和72小时显著降低(P<0.01),划痕愈合率、迁移和侵袭能力显著下降(P<0.01)(0.22±0.02)和Cyclin D1(0.21±0.02癌症。沉默LncRNA GACAT3可抑制肝细胞癌细胞的增殖、迁移和侵袭,其机制可能与抑制Wnt/β-catenin信号通路有关。关键词:长链非编码RNA胃癌相关转录物3;肝细胞癌;HepG2;Wnt/β-catenin信号通路
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Expression of long chain non coding RNA gastric cancer-associated transcript 3 in hepatocellular carcinoma and its effect on biological characteristics of HepG2 cells
Objective To investigate the expression of long chain non coding RNA (lncRNA) gastric cancer-associated transcript 3 (GACAT3) in hepatocellular carcinoma and its effect on proliferation, invasion and migration of HepG2 cells and its potential mechanism. Methods real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the differential expression of lncRNA GACAT3 in liver cancer tissue, adjacent tissues, PLC/PRF/5, Hep3B, HepG2 cell lines and normal liver cells THLE-3. Negative control small interfering RNA (siRNA) and lncRNA GACAT3 siRNA were transfected into HepG2 cells, after 48 h, Real-time PCR was used to detect the expression of lncRNA GACAT3 in transfected cells. Methyl thiazol tetrazolium (MTT) assay was used to detect cell proliferation in each group at 24, 48 and 72 h. The cell migration and invasion were detected by cell scratches experiments and Transwell assay, and the expression of Wnt/β-catenin signaling pathway protein was detected by Western blotting. Results The expression of lncRNA GACAT3 in hepatocellular carcinoma tissues (3.10±1.93) was significantly higher than that in adjacent tissues (1.02±0.59, t=8.086, P<0.01); The expression of lncRNA GACAT3 in PLC/PRF/5 (1.92±0.21, t=7.074, P<0.01), Hep3B (2.42±0.25, t=9.327, P<0.01) and HepG2 cells(2.96±0.31, t=10.582, P<0.01) were significantly higher than that in normal liver cells THLE-3 (0.97±0.10). Compared with the silencing control group (1.03±0.11), the expression of lncRNA GACAT3 in the GACAT3 group (0.16±0.02) was significantly decreased (t=13.478, P<0.01). Cell proliferation was significantly reduced at 48 h and 72 h (P<0.01). Scratch healing rate and migratory and invasive ability were significantly decreased (P<0.01). The expression of Wnt/β-catenin signaling pathway protein β-catenin (0.15±0.02) and its downstream proteins c-Myc (0.22±0.02) and Cyclin D1 (0.21±0.02) were significantly lower than that of the silencing control group(0.81±0.08, t=13.863; 0.65±0.07, t=9.731; 0.78±0.08, t=10.230; P all<0.01). The expression of p-β-catenin protein (0.75±0.08) was significantly higher than that of the silencing control group (0.27±0.03, t=11.972, P<0.01). Conclusion LncRNA GACAT3 is highly expressed in hepatocellular carcinoma. Silencing LncRNA GACAT3 can inhibit the proliferation, migration and invasion of hepatocellular carcinoma cells, and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway. Key words: Long chain non coding RNA gastric cancer-associated transcript 3; Hepatocellular carcinoma; HepG2; Wnt/β-catenin signaling pathway
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