MicroRNA-145 inhibits epithelial-mesenchymal transition in human renal cancer A-498 cells by ADAM28

AIM: To investigate the inhibitory effect of microRNA-145 (miR-145) on epithelial-mesenchymal transition (EMT) in renal cancer A-498 cells. METHODS: The A-498 cells were transfected with miR-145 mimics (M145) and mimic negative control(MNC), which served as M145 group and MNC group, respectively. Mock control (MC) group was set up using untreated A-498 cells. The expression level of miR-145 in each group was detected by RT-qPCR. Transwell assay was used to detect the invasion ability of the cells. The protein expression of vimentin, E-cadherin and ADAM28 was determined by Western blot. Bioinformatic method was used to predict the target genes of miR-145. Antagonistic effect of ADAM28 over-expression on the inhibition of EMT by miR-145 was detected by Western blot. The relationship between miR-145 and ADAM28 was analyzed by dual-luciferase reporter assay. RESULTS: The expression level of miR-145 in M145 group was significantly up-regulated than that in MC group (P<0.05). The number of invasive cells in M145 group was 12.78±3.37, which was significantly lower than that in MC group (P<0.05). ADAM28 may be the target gene of miR-145. Compared with MC group, the protein expression of vimentin and ADAM28 in M145 group was significantly decreased (P<0.05), while the protein expression of E-cadherin was significantly increased (P<0.05).After ADAM28 over-expression, the protein expression of vimentin in the A-498 cells of M145 group was significantly increased (P<0.05), and the protein expression of E-cadherin was significantly decreased (P<0.05). The results of dual-lucife-irasei reporter assay showed that ADAM28 was a downstream target gene of miR-145. CONCLUSION: miR-145 may inhibit the expression of EMT-related proteins through the downstream target gene ADAM28 and inhibit the EMT process of renal cancer A-498 cells.     

哺乳动物原始生殖细胞与EG细胞研究进展

本文综述了哺乳动物原始生殖细胞的来源、增殖、迁移及生物学特点及近年来国内外用原始生殖细胞分离EG细胞的研究进展，探讨了EG细胞作为另一种多能性干细胞的优越性。     

Effect of cocaine on caspase-3 in myocardiac cells of male rats in different age

AIM: To study the effect of cocaine on caspase-3 in myocardiac cells of male rats in different age. METHODS: Three-week-old(n=16), six-week-old(n=16) and twelve-week-old (n=16) male Sprague-Dawley rats were all divided into control groups and experiment groups randomly, each group had eight animals, experiment groups were given cocaine hydrochloride (15 mg·kg^-1 body weight) subcutaneously daily for four weeks. The ratio of heart weight to body weight (HW/BW, mg/g) were measured. DNA fragmentation of myocardia cells was determined by gel electrophoresis, and caspase-3 activity in myocardia cells was tested by flow cytometry (FCM). RESULTS: In three experiment groups, the DNA isolated from myocardial cells displayed clear ladder pattern. The HW/BW and the caspase-3 activity were increased significantly than those of control groups (P<0.01). CONCLUSIONS: Cocaine induced apoptosis in rat myocardial cells. The increase in caspase-3 activity may be the one of important pathways related to the induction of apoptosis.     

Metallothionein inhibits homocysteine-induced proliferation of rat vascular smooth muscle cells

AIM:To investigate the effect of metallothionein(MT) on proliferation of rat vascular smooth muscle cells (VSMCs) stimulated by homocysteine and its mechanism. METHODS:VSMCs proliferation was measured by [³-H]-TdR incorporation, mitogen-activated protein kinase(MAPK)activity were determined by immunoprecipitation method, the intracellular contents of MT and malondialdehyde (MDA)were assayed by -hemoglobin saturation method and TBA reaction, respectively, and lactate dehydrogenase (LDH) leakage was measured by NADH oxidation. RESULTS:Hcy(10^-6-10^-4 mmol/L) stimulated [³-H]-TdR incorporation by the VSMCs in a concentration-dependent manner. Compared with control, [³-H]-TdR incorporation in VSMCs treated with 0.1 mmol/L Hcy was increased by 4.2 fold (P＜0.01). Meanwhile, Hcy enhanced MAPK activity, MDA formation and LDH release (P＜0.01)in a concentration-dependent manner. Treatment of VSMCs with MT alone did not change above parameters, compared with control. However, MT (10^-6-10^-4 mol/L)attenuated significantly Hcy-stimulated proliferation of VSMCs (P＜0.01)in a concentration-dependent manner. And MT inhibited obviously Hcy-induced activation of MAPK activity, MDA formation and LDH release. Preincubation of VSMCs with 0.5 mmol/L ZnCl₂ for 6 h induced an increase cellular MT content by 5.7-fold (P＜0.01). The MT-overexpressed VSMCs resisted Hcy-stimulating action on MAPK activity, MDA formation and LDH leakage (P＜0.01). CONCLUSION:These results show that MT has an inhibitory effect on Hcy-induced VSMCs proliferation, and that MT could inhibit Hcy-stimulated MAPK activity and lipid peroxidation.     

Role of angiotensin II and angiotensin II receptors in vascular smooth muscle cells migration

AIM:To determine the effects of Angiotensin II(AngII) on migration of rat smooth muscle cells and to investigate the mechanisms underlying Ang II action in the development of injured vascular disease. METHODS:VSMCs isolated from aortic media of Wistar rats and cultured by the modified explant method were adopted. In prersence and absence of AngII, the expression of AngII receptor and reorganization of the actin cytoskeleton of VSMCs were studied by immunocytochemistry technique, fluorocytochemistry technique. The migration assays were performed by a modified Boyden's chamber. And the effects of AT₁R antagonist (CV-11974), AT₂R antagonist (PD123319) on aforementioned target were studied.RESULTS:VSMCs migration was stimulated by addition of AngII. The dynamic reorganization of actin cytoskeleton may be an important mechanism by which AngII facilitates VSMC motility. The expression of AT₁R in VSMCs can be upregulated after treatment with AngII initially, then decreased gradually. The expression of AT₁R was downregulated by AT₁R antagonist. The effect of AngII on VSMCs migration was mediated by AT₁R, while AT₂R had no significant effect.CONCLUSION:The dynamic reorganization of actin cytoskeleton is required for AngII-induced VSMC migration, and this effect is mediated by AT₁R .     

Comparison of different conditions inducing embryonic stem cells

AIM: To evaluate the different conditions inducing mouse embryonic stem cells (ESC) in vitro to differentiate into cardiomyocytes. METHODS: BRL conditioned medium was used to promote the growth of ESC and maintain them in an undifferentiated state. During the inducing process, retinoic acid (RA), DMSO, activin-A and TGF-β₁ were used as inducing reagents, and made up six kinds of differentiating medium. Then a three-step method inducing ESC cultured in hanging drops, in suspension and in plating was used to induce the differentiation of ESC. RESULTS: ESC were induced in vitro to differentiate into cardiomyocytes. Of all groups, the highest differentiating rate was observed in the group induced by activin-A (20 μg/L) and TGF-β₁ (2 μg/L). CONCLUSION: The inducing conditions including activin-A (20 μg/L) and TGF-β₁ (2 μg/L) is very valuable in inducing ESC differentiation into cardiomyocytes.     

Effects of berberine on IL-1 or tumour necrosis factor induced polymorphonuclear leucocyte－endothelium adhesion

AIM:To investigate the effect of berberine on IL-1 or tumour necrosis factor (TNF) induced polymorphonuclear leucocyte(PMN)-endothelium adhesion and adhesion molecules.METHODS:Based on the model of human umbilical vein endothelial cell (HUVEC), this study adopted Rose Bengal Stain, cell ELISA, immunocyto-chemical techniques to investigate the effect of berberine on PMN-endothelium adhesion and the expression of cell adhesion molecules (CAMs).RESULTS:Berberine inhibited IL-1, TNF-induced HUVEC adhesion for PMN when pretreated HUVEC and antagonised IL-1, TNF-induced upregulation of ICAM-1 on HUVEC. Meanwhile, TNF-stimulated PMN adhesion for HUVEC and CD18 upexpression on PMN was diminished in the presence of berberine.CONCLUSION: Inhibite PMN-endothelium adhesion by downregulating the CAMs expression to inhibite PMN migration across endothelium is one of the mechanisms of antiinflammation of berberine.     

Basic fibroblast growth factor and Alzheimer's disease

Alzheimer's disease (AD), which is one type of senile deimentia,has three remarkable pathologic characteristics-senile plaque, neurofibrillar tangles and neuronal death. As a neurotrophic factor, basic fibroblast growth factor (bFGF) is closely related to AD. This art icle reviewed the interaction between bFGF and AD-associated gene products (such as amyliod protein, presenilin, apoloprotein E and microtubulin-associated protein tau) and neuronal apoptosis. Some researchers suggested that bFGF may be put forward as potential therapeutic agent in AD and play a role in preventing or retarding the pathological process of this disease, alleviating or even eradicat ing the pathological insults of this disease.