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Objective To investigate the effects of Kruppel-like factor 6 (KLF6) on the apoptotic and migration ability of HepG2 cell, and the developmental role of KLF6 on zebrafish liver.Methods Constructed plasmid with shRNA-KLF6 was transfected in HepG2 and L-02.The impacts of loss of KLF6 on HepG2 was investigated by Western bolt, apoptosis analyses, cell cycle detection and scratch experiment;KLF6 morpholino oligonucleotides was microinjected into the Tg(lfabp:eGFP) transgenic zebrafish embryos.The morphant phenotype of the liver was imaged and the protein expression of KLF6 after knockdown of KLF6 was analyzed by Immunofluorescence staining.Results The expression of KLF6 in L-02 was significantly higher than in HepG2.After knockout of KLF6, KLF6 protein expression and apoptosis were significantly reduced.In addition, the cell cycle mainly stated in S phase and the migration ability of HepG2 was enhanced.After klf6 knockdown in transgenic zebrafish larvae, the development of zebrafish liver was delayed and KLF6 expression was obviously decreased in the liver.Conclusions The reduction of KLF6 expression increased the proliferation and migration ability, and reduced the apoptosis of HepG2.Loss of KLF6 affects the development of zebrafish liver, which may open a possibility to use zebrafish as a liver cancer model and for anti-liver cancer drug screening.
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Objective To research the effect of mir-520c-3p targeted GPC3 to the hepatocellular carcinoma Huh-7 cell proliferation,migration,and the influence of the attack ability and find new theoretical basis for liver hepatocellular carcinoma clinical treatment.Methods The cells were divided into three groups:not transfection of mir-520c-3p group (cell group),negative control group (Nc group),and transfection of mir-520c-3p group (treat ment group).Then used fluorescence quantitative PCR and Western Blot to detect GPC3mRNA gene and protein expression quantity.Cell proliferation of change was detected by the EDU.Made use of Transwell to detect cell invasion and migration ability of the change.Results Fluorescence quantitative PCR results showed that Cell group,NC group and treatment group were 1.13 ± 0.23,1.28 ± 0.15 and 1.05 ± 0.19 (P ＞ 0.05),mir-520-3p could not reduce the GPC3mRNA; but Western Blot detection results showed that GPC3 protein expression level reduce significantly after transfection mir-520c-3p,Cell,NC and treatment group were 2.16 ± 0.08,1.99 ± 0.04 and 0.499 ± 0.05 (P ＜ 0.01).The EDU detection results showed that hepatocellular carcinoma Huh-7 cell proliferation ability obviously inhibited after transfection mir-520c-3p,Cell group,NC group and treatment group were (90.12 ± 1.93) ％,(91.02 ± 0.35) ％ and (77.73 ± 5.88) ％ (P ＜ 0.05),and Transwell test found that hepatocellular carcinoma Huh-7cell invasion abilities were restrained,Cell group,NC group and treatment group were 0.071 ±0.008,0.105 ±0.001 and 0.048 ± 0.002 (P ＜ 0.05),in the same the cells' migration abilities were reduced,Cell group,NC group and treatment group were 0.546 ± 0.010,0.328 ± 0.002 and 0.151 ± 0.002 (P ＜0.01).Conclusions Mir-520c-3p can target GPC3 so that affect hepatocellular carcinoma Huh-7 cell proliferation,invasion and migration abilities.
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Objective To research the mechanism of impairment that cholestatic jaundice is treated as- sistly by MMDB. Methods Obstructive jaundice models with SD rats, which were divided into three time points randomly, namely 7,14,21 days,were established. In each time point, there were three groups(each six rats) : sham operation(SO + NS) ; BDL + MMDB; (BDL + NS). The rats were sacrificed at different time point, in order that liver tissue and inferior vena cava blood were taken out. The concentration of serum glutamate transaminase(AST), albumin(ALB), and total bilirubin (TB) were measured with biochemistry; the expression of TLR4mRNA was measured by reverse transcription PCR. Results On the 7th day after BDL, serum TB,AST level in BDL + MMDB and BDL + NS group were upgrade compared with SO group, in which BDL + NS group were more obviously upgrade than BDL + MMDB group(P<0.01). There was differ- ent to compare between BDL + MMDB group and SO group(P <0.01). On the 14th, 21st day after BDL, serum TB, AST coNTents were to last upgrade in BDL + MMDB and BDL + NS group. It was opposite to the SO group(P <0. 01). Conclusion The level of TLR4 could be weaken by MMDB. It may relieve the im- pairment of liver in cholestatic jaundice by LPS.
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Objective To explore the diagnosis and surgical treatment of primary intestinal tumors (PIT), to improve the diagnosis and treatment. Methods Retrospectively analysis was made on the clinical data of 72 patients with PITs admitted to our hospital from 1988 to 2002. Results Out of the 72 cases, 20.8% (15/72) had benign tumors, while the remaining 79.2% (57/72) were malignancies. The former were mostly adenoma and liomyoma, each accounting for 40.0% (6/15) of the benign tumors. Adenocarcinoma was the most common type of malignancy (36.8%,21/57), following lymphadenoma (30.0%, 17/57). The main diagnostic methods were X-ray,B-ultrasonic,CT, endoscopy and superior mesenteric arteriography.The misdingnosis rate was 62.5% before operation in this series.Of the 72 cases underwent operation,25 underwent emergent operation (33.3%, 25/72), because acute intestinal obstruction, digestive tract bleeding or perforation,acute appendicitis were diagnosed before the operation.In this series,there was no operative death;the 1,3,5 year survial rates of malignance were 62.5%,47.5%,25.0%,respectively. Conclusions PIT is not easily diagnosed before operation, and the misdiagnosis rate and emergent operation rate are high. Superior mesenteric arteriography,radiologic contrast examination of the small bowel are the important means of diagnosis in jejunum or ileum tumour, and the best way to diagnosis duodenal neoplasms is hypotonic duodenography and endoscopy. Once the diagnosis of PIT is made, the best choice of treatment is operation.