miR-126对内皮祖细胞增殖和迁移的影响以及靶基因的检测

目的 研究miR-126(micro RNA-126)对大鼠骨髓源性内皮祖细胞(endothelial progenitor cells,EPCs)增殖和迁移能力的影响,并探讨miR-126新的靶基因.方法 采用电转的方法,在EPCs中转染对照寡核苷酸和miR-126的模拟物或抑制物.噻唑蓝(MTT)法检测细胞增殖,划痕和transwell实验检测细胞迁移能力的改变.microRNA靶基因预测软件TargentScan在线分析miR-126的靶基因,并进一步用Western blot检测靶基因的表达变化.结果 (1) miR-126模拟物在转染后24 h对EPCs的增殖有促进作用(P＜0.01),转染后48和72 h,miR-126对EPCs增殖没有影响.(2)划痕和transwell实验证实miR-126模拟物可以促进EPCs的迁移(P＜0.01),miR-126抑制物抑制EPCs的迁移(P＜0.01).(3)TargetScan在线软件预测KANK2是miR-126的靶基因.(4) Western blot检测结果显示miR-126模拟物抑制KANK2的表达,miR-126抑制物促进KANK2的表达.结论 miR-126对EPCs的增殖有一过性的促进作用,miR-126可以促进EPCs的迁移能力并靶向KANK2蛋白,抑制KANK2蛋白的表达.     

Phototherapy promotes cell migration in the presence of hydroxyurea

Phototherapy has been shown to cause an increase in cell proliferation and migration. This study focused on viability (trypan blue), proliferation [sodium 3′-(1-(phenylaminocarbonyl)-3,4-tetrazolium)-bis(4-methoxy-6-nitro)-benzene sulphonic acid hydrate (XTT) and adenosine triphosphate (ATP)] and migration of WS1 cells following irradiation in the presence of hydroxyurea (HU), which is an inhibitor of proliferation. Wounded cells were irradiated on days 1 and 4 with a fluence of 5 J/cm² with a helium–neon (He-Ne) laser at 632.8 nm. After a repair time of 24 h, cellular responses were assessed. Wounded irradiated cells without HU showed an increase in cell viability and proliferation, which was confirmed by complete wound closure by day 4. Although wounded irradiated cells treated with 5 mM HU showed incomplete wound closure, these cells showed increased migration compared with that of control cells. This study showed that laser irradiation using an He-Ne laser with a fluence of 5 J/cm² stimulates cell viability. The HU results confirmed that laser irradiation promotes cell migration and proliferation.     

In vitro inhibition of lens epithelial cell proliferation and migration

Delayed opacification of the posterior capsule is the most common cause of decreased visual acuity after extracapsular cataract extraction. In humans, this has been shown to result from migration and possibly proliferation of residual lens epithelial cells onto the central posterior capsule. We studied a group of pharmacologic agents to determine their ability to inhibit lens epithelial cell proliferation and migration in vitro. A drug capable of inhibiting lens epithelial cell growth and/or migration, if free of unacceptable toxic effects on other cell populations, might be used to prevent lens capsule opacification. Anti-proliferative activity was exhibited by several agents, with 50% inhibition of growth occurring at the following concentrations: 5-fluorouracil, 30 micrograms/ml; daunomycin, 10 ng/ml; colchicine, 15 ng/ml; doxorubicin, 5 ng/ml; dexamethasone, 100 micrograms/ml; and cytosine arabinoside, 100 ng/ml. Colchicine inhibited lens epithelial cell migration by 50% at 20 ng/ml.     

NF-kappaB regulates intestinal epithelial cell and bile salt-induced migration after injury

OBJECTIVE: To determine if NF-kappa B regulates intestinal epithelial cell migration and if it has a role during bile salt-induced migration. SUMMARY BACKGROUND DATA: Mucosal restitution is an important repair modality in the gastrointestinal tract. The authors have shown that taurodeoxycholate (TDCA) increases intestinal epithelial cell migration. NF-kappa B regulates activation of a number of genes involved in inflammatory responses. METHODS: Studies were conducted in IEC-6 cells. I kappa B protein expression was determined by Western blot analysis. Sequence-specific NF-kappa B binding activity was measured by EMSA shift assays and nuclear localization by immunohistochemistry. Cell migration was examined by using an in vitro model that mimics the early cell division-independent stages of epithelial restitution. RESULTS: The process of cell migration over the wounded area was associated with a significant increase in NF-kappa B binding activity in IEC-6 cells. Immunohistochemistry revealed translocation of NF-kappa B into the nucleus. Western blot analysis showed that injury decreased I kappa B protein expression. Inhibition of the binding activity by treatment with a specific NF-kappa B inhibitor, MG-132, inhibited cell migration during restitution. Further, exposure to TDCA at the physiologic concentration that induces intestinal epithelial cell migration increased NF-kappa B binding activity, induced NF-kappa B translocation into the nucleus, and decreased I kappa B protein expression. MG-132 also inhibits bile salt-induced cell migration. CONCLUSIONS: NF-kappa B regulates intestinal epithelial cell migration. Bile salts at physiologic concentrations increase cell migration by activation of NF-kappa B. These data show that bile salts may have a role in the maintenance of intestinal mucosal integrity.     

High-frequency low-level diode laser irradiation promotes proliferation and migration of primary cultured human gingival epithelial cells

In periodontal therapy, the use of low-level diode lasers has recently been considered to improve wound healing of the gingival tissue. However, its effects on human gingival epithelial cells (HGECs) remain unknown. The aim of the present study was to examine whether high-frequency low-level diode laser irradiation stimulates key cell responses in wound healing, proliferation and migration, in primary cultured HGECs in vitro. HGECs were derived from seven independent gingival tissue specimens. Cultured HGECs were exposed to a single session of high-frequency (30 kHz) low-level diode laser irradiation with various irradiation time periods (fluence 5.7–56.7 J/cm²). After 20–24 h, cell proliferation was evaluated by WST-8 assay and [³H]thymidine incorporation assay, and cell migration was monitored by in vitro wound healing assay. Further, phosphorylation of the mitogen-activated protein kinase (MAPK) pathways after irradiation was investigated by Western blotting. The high-frequency low-level irradiation significantly increased cell proliferation and [³H]thymidine incorporation at various irradiation time periods. Migration of the irradiated cells was significantly accelerated compared with the nonirradiated control. Further, the low-level diode laser irradiation induced phosphorylation of MAPK/extracellular signal-regulated protein kinase (ERK) at 5, 15, 60, and 120 min after irradiation. Stress-activated protein kinases/c-Jun N-terminal kinase and p38 MAPK remained un-phosphorylated. The results show that high-frequency low-level diode laser irradiation promotes HGEC proliferation and migration in association with the activation of MAPK/ERK, suggesting that laser irradiation may accelerate gingival wound healing.     

Aquaporin-1 facilitates epithelial cell migration in kidney proximal tubule

Aquaporin-1 (AQP1) is the principal water-transporting protein in cell plasma membranes in kidney proximal tubule, where it facilitates transepithelial water transport. Here, a novel role for AQP1 in kidney involving the migration of proximal tubule cells is reported. Migration was compared in primary cultures of proximal tubule cells from wild-type and AQP1 null mice. Cell cultures from AQP1 null mice were indistinguishable from those of wild-type mice in their appearance, growth/proliferation, and adhesiveness, although, as expected, they had reduced plasma membrane water permeability. Migration of AQP1-deficient cells was reduced by >50% compared with wild-type cells, as measured in a Boyden chamber in the presence of a chemotactic stimulus. Comparable slowing of migration of AQP1-deficient cells was also found in an in vitro scratch assay of wound healing, with reduced appearance of lamella-like membrane protrusions at the cell leading edge. Adenoviral-mediated expression of AQP1 in the AQP1-deficient cells, which increased their water permeability to that of wild-type cells, corrected their migration defect. The potential relevance of these in vitro findings to the intact kidney was tested in an in vivo model of acute tubular injury caused by 30 min of renal artery occlusion. At 3 to 5 d after ischemia-reperfusion, kidneys in AQP1 null mice showed remarkably greater tubular injury and cellular actin disorganization than kidneys in wild-type mice. These results provide evidence for the involvement of AQP1 in migration of proximal tubule cells and possibly in the response of the proximal tubule to injury.     

RPS7 promotes cell migration through targeting epithelial-mesenchymal transition in prostate cancer

Objectives

Small ribosomal protein subunit 7 (RPS7) is an important structural components of the ribosome involved in protein synthesis, previous studies demonstrated that RPS7 was associated with several malignancies, but the role of RPS7 in prostate cancer (PCa) remains unclear. To decipher such a puzzle, in the current study, we deciphered the role and mechanism of RPS7 during the progression of PCa.

Bile salt stimulates intestinal epithelial cell migration through TGFbeta after wounding

BACKGROUND: In addition to aiding in the digestion of fats, luminal bile salts have been shown to modulate gastrointestinal epithelial growth, differentiation, and other functions. We hypothesized that bile acids could modulate the intestinal mucosal repair process of restitution. We investigated the effect of the bile salt taurodeoxycholic acid on epithelial migration and identified a role for TGFbeta, a widely expressed cytokine in the intestinal villus, in this repair process. METHODS: Using a well-established model of epithelial restitution, IEC-6 cells were plated on 60-mm Matrigel-coated plastic dishes and grown to confluence. The epithelium was wounded by scraping with a 6-mm-wide blade to create a smooth denuded edge and cell migration was measured 8 h later. Cells were grown in control DMEM with 5% FBS with or without 0.01-2 mM taurodeoxycholic acid (TDCA). In parallel experiments, cells were harvested for Northern analysis of TGFbeta and GAPDH expression; [3H]thymidine uptake was used to measure proliferation. Anti-TGFbeta antibody was added to cells grown in the presence of 0.05 mM TDCA and migration was measured at 8 h. RESULTS: TDCA at physiologic luminal concentrations augments IEC-6 cell migration, with a maximal effect at 0.05 mM. TDCA inhibited proliferation at these concentrations. TGFbeta expression increased in response to bile acid, while wounding had less of an effect on TGFbeta expression. Blockade of TGFbeta function with TGFbeta antibody eliminated the effect of bile on cell migration. CONCLUSIONS: Bile acid at physiologic concentrations augments small intestinal epithelial cell migration. The process is dependent on TGFbeta and is independent of cell division. The data further support a role for bile acids and TGFbeta in differentiated intestinal cell function and in preservation of an intact mucosa.     

circZBTB44通过调控AKT通路促进肾透明细胞癌增殖和迁移的研究

目的 探究circZBTB44在肾透明细胞癌中的表达情况及探讨circZBTB44在肾透明细胞癌中的生物学功能及促进肾透明细胞癌细胞进展的可能机制.方法 通过qRT-PCR法检测circZBTB44在我院21对肾透明细胞癌组织及细胞系786-O、ACHN中的表达水平;通过放线菌素D实验、RNase R实验、核浆分离实验...     

Nicotine promotes cell migration through alpha7 nicotinic acetylcholine receptor in gastric cancer cells

Background  

The objective was to study the mechanism of nicotine-enhanced migration of gastric cancer cells. Long-term cigarette smoking increases the risk of gastric cancer mortality. Tobacco-specific mitogen, nicotine, was reported to correlate with cancer progression on gastric cancer. Since metastasis is the major cause of cancer death, the influence of nicotine on the migration of gastric cancer cells remains to be determined.     

Heparin-binding EGF-like growth factor (HB-EGF) promotes cell migration and adhesion via focal adhesion kinase

Background

Cell migration and adhesion are essential in intestinal epithelial wound healing and recovery from injury. Focal adhesion kinase (FAK) plays an important role in cell–extracellular matrix signal transduction. We have previously shown that heparin-binding EGF-like growth factor (HB-EGF) promotes intestinal epithelial cell (IEC) migration and adhesion in vitro. The present study was designed to determine whether FAK is involved in HB-EGF–induced IEC migration and adhesion.

Materials and methods

A scrape wound healing model of rat IECs was used to examine the effect of HB-EGF on FAK-dependent cell migration in vitro. Immunofluorescence and Western blot analyses were performed to evaluate the effect of HB-EGF on the expression of phosphorylated FAK (p-FAK). Cell adhesion assays were performed to determine the role of FAK in HB-EGF–induced cell adhesion on fibronectin (FN).

Results

HB-EGF significantly increased healing after scrape wounding, an effect that was reversed in the presence of an FAK inhibitor 14 (both with P < 0.05). HB-EGF increased p-FAK expression and induced p-FAK redistribution and actin reorganization in migrating rat IECs. Cell adhesion and spreading on FN were significantly increased by HB-EGF (P < 0.05). FAK inhibitor 14 significantly inhibited both intrinsic and HB-EGF–induced cell adhesion and spreading on FN (both with P < 0.05).

Conclusions

FAK phosphorylation and FAK-mediated signal transduction play essential roles in HB-EGF–mediated IEC migration and adhesion.     

Aquaporin 2 mutations in nephrogenic diabetes insipidus

Water reabsorption in the renal collecting duct is regulated by the antidiuretic hormone vasopressin (AVP). When the vasopressin V2 receptor, present on the basolateral site of the renal principal cell, becomes activated by AVP, aquaporin-2 (AQP2) water channels will be inserted in the apical membrane, and in this fashion, water can be reabsorbed from the pro-urine into the interstitium. The essential role of the vasopressin V2 receptor and AQP2 in the maintenance of body water homeostasis became clear when it was shown that mutations in their genes cause nephrogenic diabetes insipidus, a disorder in which the kidney is unable to concentrate urine in response to AVP. This review describes the current knowledge on AQP2 mutations in nephrogenic diabetes insipidus.     

类风湿关节炎中巨噬细胞移动抑制因子对内皮细胞功能及血管内皮生长因子的影响

目的探讨巨噬细胞移动抑制因子（MIF）对类风湿关节炎（RA）滑膜内皮血管生成的影响和作用机制。方法免疫组化检查MIF在RA滑膜组织中的表达;加入MIF或PBS溶液培养人微血管内皮细胞株（HMEC-1）,CCK8法观察细胞的生长差异,磷脂酰丝氨酸凋亡试剂盒（AnnexinV法）检测细胞凋亡,RT—QPCR检测细胞中MIF及VEGF的表达差异。结果MIF在5例RA组织中均阳性表达;加入MIF的细胞株增殖显著较加PBS溶液的快（F=216．93,P〈0．01）,细胞株的凋亡被MIF显著抑制（凋亡率从21．37％降为7．01％（t=13．88,P〈0．01）;处于分裂相的细胞数目较加入PBS的细胞株多（G2期＋S期细胞比例从37．89％升为54．05％,t=5．42,P〈0．01）,在加入MIF的细胞株中VEGF的表达4．62倍高于加入PBS的细胞株（t=7．34,P〈0．01）。结论MIF在RA滑膜组织中表达,并通过提高血管内皮细胞的VEGF而促进RA滑膜血管生成而发挥重要的生理作用。MIF或许是治疗RA进展的药物靶点,有望通过抑制RA患者中MIF的表达从而延缓疾病的进展,改善患者的生活质量。     

UBE2W down-regulation promotes cell apoptosis and correlates with hypospermatogenesis

Ubiquitin conjugating enzyme (E2) is crucial for mediating N-terminal ubiquitination. Recent study reports that UBE2W is involved in male infertility. However, the correlation between UBE2W expression and hypospermatogenesis is unclear. The present study is to explore the biological role of UBE2W and its association with hypospermatogenesis. Results showed that the sexpression levels of UBE2W in mouse testes were gradually elevated from 2 to 10 weeks, while were significantly deceased in the testes with hypospermatogenesis. When UBE2W expression was successfully down-regulated in spermatogenic cells, the rate of apoptosis was significantly increased and the P53/Bcl-2/caspase 6/caspase 9 signal pathways were activated. Thus, these data indicate that UBE2W down-regulation promotes cell apoptosis and correlates with hypospermatogenesis, which may be helpful for the diagnosis of male infertility.     

Ischemia-reperfusion of small liver remnant promotes liver tumor growth and metastases--activation of cell invasion and migration pathways.

Elucidating the mechanism of liver tumor growth and metastasis after hepatic ischemia-reperfusion (I/R) injury of a small liver remnant will lay the foundation for the development of therapeutic strategies to target small liver remnant injury, and will reduce the likelihood of tumor recurrence after major hepatectomy or liver transplantation for liver cancer patients. In the current study, we aimed to investigate the effect of hepatic I/R injury of a small liver remnant on liver tumor development and metastases, and to explore the precise molecular mechanisms. A rat liver tumor model that underwent partial hepatic I/R injury with or without major hepatectomy was investigated. Liver tumor growth and metastases were compared among the groups with different surgical stress. An orthotopic liver tumor nude mice model was used to further confirm the invasiveness of the tumor cells from the above rat liver tumor model. Significant tumor growth and intrahepatic metastasis (5 of 6 vs. 0 of 6, P=0.015), and lung metastasis (5 of 6 vs. 0 of 6, P=0.015) were found in rats undergoing I/R and major hepatectomy compared with the control group, and was accompanied by upregulation of mRNA levels for Cdc42, ROCK (Rho kinase), and vascular endothelial growth factor, as well as activation of hepatic stellate cells. Most of the nude mice implanted with liver tumor from rats under I/R injury and major hepatectomy developed intrahepatic and lung metastases. In conclusion, hepatic I/R injury of a small liver remnant exacerbated liver tumor growth and metastasis by marked activation of cell adhesion, invasion, and angiogenesis pathways.