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Lamin B1 (LMNB1) is highly expressed in liver cancer tissues, and its influence and mechanism on the proliferation of hepatocellular carcinoma cells were explored by knocking down the expression of the protein. In liver cancer cells, siRNAs were used to knock down LMNB1. Knockdown effects were detected by Western blotting. Changes in telomerase activity were detected by telomeric repeat amplification protocol assay (TRAP) experiments. Telomere length changes were detected by quantitative real-time polymerase chain reaction (qPCR). CCK8, cloning formation, transwell and wound healing were performed to detect changes in its growth, invasion and migration capabilities. The lentiviral system was used to construct HepG2 cells that steadily knocked down LMNB1. Then the changes of telomere length and telomerase activity were detected, and the cell aging status was detected by SA-β-gal senescence staining. The effects of tumorigenesis were detected by nude mouse subcutaneous tumorigenesis experiments, subsequent histification staining of tumors, SA-β-gal senescence staining, fluorescence in situ hybridization (FISH) for telomere analysis and other experiments. Finally, the method of biogenesis analysis was used to find the expression of LMNB1 in clinical liver cancer tissues, and its relationship with clinical stages and patient survival. Knockdown of LMNB1 in HepG2 and Hep3B cells significantly reduced telomerase activity, cell proliferation, migration and invasion abilities. Experiments in cells and tumor formation in nude mice had demonstrated that stable knockdown of LMNB1 reduced telomerase activity, shortened telomere length, senesced cells, reduced cell tumorigenicity and KI-67 expression. Bioinformatics analysis showed that LMNB1 was highly expressed in liver cancer tissues and correlated with tumor stage and patient survival. In conclusion, LMNB1 is overexpressed in liver cancer cells, and it is expected to become an indicator for evaluating the clinical prognosis of liver cancer patients and a target for precise treatment.
Subject(s)
Animals , Mice , Telomerase/metabolism , Carcinoma, Hepatocellular/genetics , Liver Neoplasms/genetics , Telomere Shortening , In Situ Hybridization, Fluorescence , Mice, Nude , Telomere/pathology , CarcinogenesisABSTRACT
Objective:To explore the classification and treatment strategy of femoral neck fracture combined with ipsilateral intertrochanteric fracture.Methods:A retrospective analysis was performed of the 44 patients who had been admitted to Department of Orthopedics, Liangxiang Teaching Hospital, Capital Medical University from March 2003 to March 2019 for femoral neck fracture combined with ipsilateral intertrochanteric fracture. They were 19 males and 25 females, aged from 37 to 93 years (average, 77.9 years). According to the anatomical location and displacement severity, the femoral neck fractures were divided into 3 types while the intertrochanteric fractures were classified as stable or unstable ones. There were 3 cases of type Ⅰ which were completely extracapsular ones, 31 cases of type Ⅱ which were intracapsular stable ones, and 10 cases of type Ⅲ which were intracapsular unstable ones. Types Ⅰ and Ⅱ fractures were treated with intramedullary fixation, and type Ⅲ fractures with cemented hip hemi-replacement+reduction and fixation of the intertrochanteric fracture with Kirschner wires and steel cables. Recorded were fracture healing time, function of the affected hip and complications.Results:The 44 patients were followed up for at least 2 years. The fracture healing time for the 3 patients with type I fracture averaged 5.6 months (from 4.4 to 6.8 months); their hip function at the last follow-up, evaluated by the Harris scoring system, was excellent in 2 cases and good in one case. For 30 of the 31 patients with type Ⅱ fracture, the fracture healing time averaged 7.2 months (from 5.1 to 9.3 months); their hip function at the last follow-up, evaluated by the Harris scoring system, was excellent in 18 cases, good in 6 cases, fair in 5 cases, and poor in 2 cases, giving an excellent and good rate of 77.4% (24/31). As for complications, withdrawal or cutting-out of the head screw happened in 6 cases, infection in one case and nonunion in one case. In 10 patients with type Ⅲ fracture, the hip function at the last follow-up was excellent in 7 cases, good in 2 cases and poor in one case.Conclusions:For type Ⅰ and type Ⅱ femoral neck fractures combined with ipsilateral intertrochanteric fracture, intramedullary fixation with angulation stability may be a proper choice. For type Ⅲ ones, hip joint replacement should be the first choice.
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BACKGROUND:Previous studies have shown that monocytes can transdifferentiate into vascular endothelial cells under the induction of various factors including vascular endothelial growth factor(VEGF).It remains poorly understood whether monocytes can be induced to transdifferentiate into lymphatic endothelial cells in vitro.OBJECTIVE:To explore the possibility of the transdifferentiation of monocytes into lymphatic endothelial cells under inflammatory condition.METHODS:Fresh monocytes from peripheral blood were collected by Ficoll density gradient centrifugation and cultured in an endothelial cell medium,followed by incubation in fibronectin-plated well or treated with tumor necrosis factor a for 24 hours,respectively.The expression of specific markers of lymphatic endothelial cells,such as LYVE-1,Podoplanin,Porx-1 and VEGF receptor 3(VEGFR-3),as well as the endothelial cells markers,such as vWF,endothelial nitric oxide synthase(eNOS)and VEGFR-2,were detected by RT-PCR and immunochemical methods.RESULTS AND CONCLUSION:Prior to induction,monocytes were positive to LYVE-1,but negative for Podoplanin,Porx-1,and VEGFR-3,vWF,eNOS,as well as VEGFR-2.Following induction,the cultured mononcytes were positive for Podoplanin,Prox-1 and VEGFR-3,but remained negative for vWF,eNOS and VEGFR-2.It suggested that monocytes can be induced to express the markers of lymphatic endothelial cells stimulated by fibronectin or tumor necrosis factor a.
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Objective: To study the influence of inflammatory cytokine TNF-? on the expression of adhesion molecules and specific markers of rat MSCs, and to study the optimal stimulation of MSCs with inflammatory factors in inducing adhesion molecule expression which promotes migration of MSCs to the ischemic area in liver. Methods: The MSCs stimulated with different concentration of TNF-? were detected for adhesion molecules and stem cells markers on cell surface with the method of flow-cytometry, MSCs which were stimulated with the optimal concentration of TNF-? and labeled with 1, 1-Dioctadecyl-3, 3, 3, 3-tetramethylindocarbocyanine Iodade(DiI)were delivered intravenously to the rats whose liver was injured by ischemia, the liver function of the experimented animals were tested, and liver samples in the ischemic area were obtained, the number of MSCs was counted under a fluorescent microscope. Results: Stimulated with TNF-?, MSC ex-pression of VCAM-1 increased, while that of stem cell markers did not change markedly. Exposed to lower concentration of TNF-?, the adhesion ability of MSCs obviously increased and more MSCs rested in the ischemic areas in the rat liver, compared to the control groups. Conclusions: TNF-? can increase the expression of VCAM-1 on rat MSCs while exert little effect on the stem cell character of MSCs. Suitable concentration of TNF-? can promote MSCs migration to the damaged tissue, which provides rationale for the treatment of liver disease.