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目的研究肝泡型包虫病患者肝脏细胞Gadd45β(growth arrest and DNA damage gene 45β)及细胞周期调控相关基因的表达及意义。方法采集24例肝泡型包虫病患者肝脏标本,分为病灶旁组织(Close)和远端组织(Distance)。用重组TGF-β1(1ng/ml)及泡球蚴匀浆蛋白(1mg/ml)刺激体外培养的HL-7702人肝细胞,分别于30min、1h、6h、12h、24h、48h收集细胞。患者肝脏标本及刺激后收集的细胞均采用Trizol法提取RNA,反转录cDNA,通过实时荧光定量PCR检测目的基因表达并进行分析。结果近旁组织PCNA,TGF-β1,cyclinA1,cyclinE基因相对表达量分别为1.934±1.337、1.155±0.5138、1.367±0.8087、1.384±1.116、1.580±1.005、1.899±1.173和1.478±1.189,显著高于远端组织(1.00±0.00)且差异有统计学意义(P<0.05),而Gadd45β,cyclinB1,cyclinD差异无统计学意义(P>0.05)。用重组TGF-β1(1ng/ml)及泡球蚴匀浆蛋白(1mg/ml)刺激体外培养的HL-7702人肝细胞,引起Gadd45β,cyclinA1,cyclinE表达上调。结论泡球蚴匀浆蛋白刺激,可引起肝细胞Gadd45β及细胞周期相关蛋白表达上调,参与细胞抗凋亡过程。泡球蚴感染晚期,肝脏中仍存在细胞周期素(cyclins)参与的细胞增殖,但同时TGF-β1高表达,并通过Smads信号通路促进肝纤维化相关蛋白表达,促成肝脏泡型包虫病晚期病理变化。
Objective To study the expression and significance of Gadd45β (hepatocyte growth arrest and DNA damage gene 45β) and cell cycle related genes in patients with hepatic echinococcosis. Methods Liver samples of 24 patients with hepatic echinococcosis were collected, which were divided into two groups: Close and distant. HL-7702 human hepatocytes cultured in vitro were stimulated with recombinant TGF-β1 (1 ng / ml) and cysticercus cellulolytic protein (1 mg / ml), and the cells were harvested at 30 min, 1 h, 6 h, 12 h, The Trizol method was used to extract the RNA from the liver samples and the cells after stimulation. The cDNAs were reverse transcribed and the expression of the target gene was detected by real-time fluorescence quantitative PCR. Results The relative expression of PCNA, TGF-β1, cyclinA1 and cyclinE were 1.934 ± 1.337, 1.155 ± 0.5138, 1.367 ± 0.8087, 1.384 ± 1.116, 1.580 ± 1.005, 1.899 ± 1.173 and 1.478 ± 1.189 respectively, which were significantly higher than those of far (1.00 ± 0.00) and the difference was statistically significant (P <0.05), while there was no significant difference between Gadd45β, cyclinB1 and cyclinD (P> 0.05). The HL-7702 human hepatocytes cultured in vitro were stimulated with recombinant TGF-β1 (1 ng / ml) and cysticercus cellulosae homogenate (1 mg / ml) to induce the up-regulation of Gadd45β, cyclinA1 and cyclinE. Conclusion Cyclospora cercariae protein stimulation can induce the up-regulation of Gadd45β and cell cycle-related proteins in hepatocytes, which is involved in the anti-apoptotic process of cells. Cysticercosis late infection, there are still liver cells in the cyclins (cyclins) involved in cell proliferation, but at the same time TGF-β1 expression, and through the Smads signaling pathway to promote liver fibrosis-related protein expression, promote liver hydatid disease late Pathological changes.