1.The influence of HGF on the expression of VEGF-C and its mechanism in human tongue squamous cell carcinoma Tca8113 cells
Hongliang DU ; Dengqi HE ; Zhirui ZHANG ; Jilian XU ; Yaming XI
Journal of Practical Stomatology 2014;(6):792-795
Objective:To investigate the influence of hepatocyte growth factor(HGF)on the expression of vascular endothelial growth factor C(VEGF-C)and the mechanism of HGF-induced VEGF-C expression in tongue squamous cell carcinoma Tca8113 cells.Methods:Tca8113 cells were cultured and exposed to HGF with various concentrations.The expression level of VEGF-C was assessed by ELISA.Signaling transduction inhibitors LY294002,U0126,SP600125,SB203580 was used to block PI3K/Akt,P44 /P22MAPK,JNK,P38MAPK signaling pathways,respectively.Then,the expression level of VEGF-C was detected by ELISA.Re-sults:The VEGF-C expression of Tca8113 cells increased at the beginning and decreased later with the increase of HGF concentra-tion.When the concentration of HGF was 40 ng/ml,VEGF-C expression level was the highest.Inhibitor LY294002 of PI3K/Akt and Inhibitor U0126 of P44 /P22MAPK significantly blocked the effects on HGF-induced VEGF-C up-regulation(P <0.01 ).Inhibitor SP600125 of JNK and inhibitor SB203580 of P38MAPK didn't interfere HGF-induced VEGF-C expression(P >0.05).Conclusion:HGF contributed to the expression of VEGF-C,PI3K/Akt and P44 /P22MAPK signaling pathways may be involved in HGF-induced VEGF-C up-regulation,and may play potential roles in lymphatic metastasis of oral squamous cell carcinoma.
2.Identification and in vitro BrdU labeling of adult bone marrow mesenchymal stem cells
Xiaorui WANG ; Xianglin HOU ; Yaming XI ; Haihong ZHANG
Chinese Journal of Tissue Engineering Research 2007;0(21):-
BACKGROUND: No particular marker molecule of bone marrow mesenchymal stem cells (BMSCs) is presently found, so its determination is difficult. BrdU marker has no radioactive pollution. Some studies have confirmed that BrdU marker has no damage to cell function without ultraviolet radiation. OBJECTIVE: To investigate in vitro identification and labeling methods and biological characteristics of adult BMSCs. DESIGN, TIME AND SETTING: The cell experiment was conducted at the General Hospital of Lanzhou Military Area Command of Chinese PLA between June and December 2007. MATERIALS: Bone marrow was collected from 5 healthy adult volunteers. BrdU was purchased from Sigma, USA. METHODS: 10 mL adult bone marrow was harvested to isolate mononuclear cells by density gradient. Cells were cultured in DMEM containing 10% fetal bovine serum and proliferated at a density of 2?108 L-1. At the third passage, BMSCs was inoculated in medium supplemented with osteoblast inductor and stained with alkaline phosphatase. Subsequently, BMSCs were inoculated in medium supplemented with lipoblast inductor and stained with oil red O. Cells were incubated with BrdU at different concentrations of 5, 10 and 15 ?mol/L for 12, 24, 48, 72 and 96 hours, and then detected by immunocytochemistry. MAIN OUTCOME MEASURES: Cell growth and proliferation were observed under an inverted light microscope. Cell phenotype, osteoblast and lipoblast differentiation were identified by flow cytometry. BrdU-positive labeling rate and cell growth after labeling were investigated. RESULTS: In vitro cultured BMSCs were homogenous population and exhibited a spindle-shaped fibroblastic morphology. BMSCs expressed CD44 and CD71, but did not express CD34 and HLA-DR. BMSCs can differentiate into osteoblasts and lipoblasts. Most BMSCs were labeled by BrdU. With the increase in labeling concentration and over time, BrdU positive rate gradually increased and exceeded 90% after labeling with BrdU (10 ?mol/L) for 72 hours, with the labeling identifiable in nine consecutive passages. At the third passage, 90% BMSCs were in G0/G1 phase, and 88.62% in G0/G1 phase after labeling. BrdU has little effect on morphous and proliferation of labeled cells. CONCLUSION: Adult BMSCs can be identified through morphological character, specific surface antigens and multipotential differentiation in vitro. BrdU labeling provides a feasible means for a dynamic observation of the survival, growth and differentiation of the implanted BMSCs. The optimal dosage and timing of BrdU labeling are respectively 10 ?mol/L and 72 hours.
3.A relationship between genetic polymorphisms of CYP2E1 and susceptibility of acute leukemia in Gansu population
Xiue SHI ; Jianwang XU ; Jialing MA ; Hai LIU ; Mingfeng JIA ; Chunxia LIU ; Yaming XI
Cancer Research and Clinic 2010;22(z1):5-7
Objective To study the relationship between genetic polymorphisms of CYP2E1 and the susceptibility of the acute leukemia in Gansu population. Methods The C609T polymorphism of CYP2E1 gene was detected by polymerase chain reaction-ligase detection reaction (PCR-LDR) and 1∶1 matched casecontrol method in 100 healthy persons (control group) and 100 patients with acute leukemia (AL group).Results The C2 allele genotype and C1C2/C2C2 genotype of CYP2E1 gene occurred more frequently in AL group (13.5 % and 22 %, respectively) than those in control group (10.5 % and 19 %, respectively), however,both differences showed no statistical significant. Further stratified analysis, the C1C2/C2C2 genotype of CYP2E1 gene occurred more frequently in AML group (27%) than that in control group (19 %), but difference had no statistical significant, too. The occurrence frequency of the C2 allele genotype and C1C2/C2C2 genotype of CYP2E1 gene showed no significant difference in ALL group and control group (x2=0.446, P =0.504>0.05). Conclusion Genetic polymorphisms of CYP2E1 don't correlated to susceptibility of acute leukemia(AML and ALL) in Gansu population.
4.Analyses of T-lymphocyte rDNA transcription in peripheral blood of patients with stomach-intestine tract malignant tumor
Yaming XI ; Bei SUN ; Huaxi WANG ; Jinyi LI ; Yihao XU ; Yunqi SU
Chinese Journal of Pathophysiology 2000;0(07):-
AIM: To examine T-lymphocyte rDNA transcription activity in peripheral blood of patients with gastrointestinal maliganant tumor and to clarify its clinical significance.METHODS: T-lymphocyte rDNA transcription activity in peripheral blood of 48 cases of patients with stomach-intestine tract malignant tumor were measured.RESULTS: Before surgery, the T-lymphocyte rDNA transcviption activity was obviously lower than that after surgery, also lower than that of the normal control( P