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PURPOSE: Although the effect of lysosome-associated protein transmembrane 4 beta (LAPTM4B) on the proliferation, migration, and invasion of breast cancer (BC) cells has already been studied, its specific role in BC progression is still elusive. Here, we evaluated the effect of different levels of LAPTM4B expression on the proliferation, invasion, adhesion, and tumor formation abilities of BC cells in vitro, as well as on breast tumor progression in vivo. METHODS: We investigated the influence of LAPTM4B expression on MCF-7 cell proliferation, invasion, adhesion, and tube formation abilities in vitro through its overexpression or knockdown and on breast tumor progression in vivo. RESULTS: Cell growth curves and colony formation assays showed that LAPTM4B promoted the proliferation of breast tumor cells. Cell cycle analysis results revealed that LAPTM4B promoted the entry of cells from the G1 into the S phase. Transwell invasion and cell extracellular matrix adhesion assays showed that LAPTM4B overexpression increased the invasion and adhesion capabilities of MCF-7 cells. More branches were observed in MCF-7 cells overexpressing LAPTM4B under an electron microscope. In comparison with LAPTM4B overexpression, LAPTM4B knockdown decreased the expression of vascular endothelial growth factor-A and significantly inhibited the vasculogenic tube formation ability of tumors. These results were also verified with western blot analysis. CONCLUSION: LAPTM4B promoted the proliferation of MCF-7 cells through the downregulation of p21 (WAF1/CIP1) and caspase-3, and induced cell invasion, adhesion, and angiogenesis through the upregulation of hypoxia-inducible factor 1 alpha, matrix metalloproteinase 2 (MMP2), and MMP9 expression. This specific role deems LAPTM4B as a potential therapeutic target for BC treatment.
Subject(s)
Blotting, Western , Breast Neoplasms , Breast , Caspase 3 , Cell Cycle , Disease Progression , Down-Regulation , Extracellular Matrix , Hypoxia-Inducible Factor 1 , In Vitro Techniques , Matrix Metalloproteinase 2 , MCF-7 Cells , S Phase , Up-Regulation , Vascular Endothelial Growth Factor AABSTRACT
Endothelial to mesenchymal transition(EndMT) plays a major role during organism development, and also contributes to several adult cardiovascular diseases.EndMT-derived fibroblast-like cells are common in atherosclerotic lesions.Pro-atherosclerosis factors, such as oxidative stress, hypoxia, inflammatory cytokines and oscillatory fluid shear stress can promote EndMT.EndMT is closely associated with plaque calcification, and unstable and ruptured plaque phenotype that may prone to cause clinical events.EndMT may be another key step in the prevention and treatment of atherosclerosis.Here, we reviewed the role played by endothelial-to-mesenchymal transition(EndMT) and its key regulators in atherosclerosis.
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Aim To study the effect of Tongxinluo on the polarization of macrophages to type M2 and M1, and on Notch signaling pathway.Methods THP-1 was respectively induced by PMA and IL-4 or LPS to M2 and M1 type polarized macrophages, and treated by Tongxinluo.The expressions of marker molecules IL-10, TNF-α, IL-6 and protein of Notch signaling pathway were detected.Results The level of IL-10 increased in M2 group and there was no statistical difference on the level of IL-6 and TNF-α;the level of IL-10 decreased and IL-6, TNF-α increased in M1 group;the level of IL-10 increased and IL-6,TNF-α decreased in TXL group.There was no statistical difference in the expression of active Notch-1, DLL4 and Hes-1 between control group and M2 group;the expression of active Notch-1, DLL4 and HES-1 increased significantly in M1 group;the expression of active Notch-1, DLL4 and HES-1 decreased significantly in TXL group;there was no significant difference in the expression of Notch-1 protein in each group.Conclusion The polarization of macrophages to M1 type can be inhibited by Tongxinluo, and its mechanism may be related to the inhibition of activation of Notch-1 signaling pathway.
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Objective: To observe the effect of oxidative-low density lipoprotein (ox-LDL) injured human leukemia mononuclear cells (THP-1) adhesion to human umbilical vein endothelial cells (HUVECs)in vitrowith the intervening function of dredging collateral drug, tongxinluo (TXL) and ginsenoside (Rb1). Methods: Cell injury was induced by ox-LDL treatment. The cells were divided into 4 groups:①Normal control group,②Injury model group, the cells were cultured by ox-LDL,③TXL group, the cells were cultured with both ox-LDL and TXL,④Rb1 group. HUVEC viability was measured by MTS assay, adherence rate of THP-1 cells to HUVECs was tested by vital cell staining. The contents of monocyte chemoat-tractant protein (MCP-1), soluble vascular cell adhesion molecule (sVCAM-1), soluble inter vascular cell adhesion molecule-1 (sICAM-1) and E-selectin in HUVEC conditioned medium were detected by ELISA; protein expressions of CCR2, VLA4 and Mac-1 in THP-1 cells were examined by Western blot analysis. Results: Compared with Control group, HUVEC viability was decreased in Injury model group (100 ±1.31) % vs(75.57 ± 1.02) %, while increased in both TXL and Rb1 groups (99.25 ± 1.40) % and (99.48 ± 2.15) %; Injury model group showed elevated adherence rate of THP-1 cells to HUVECs, while the adherence rates were reduced in both TXL and Rb1 groups. Compared with Injury model group, TXL group and Rb1 group showed decreased levels of MCP-1, sVCAM-1, sICAM-1 and E-selectin in HUVEC conditioned medium; decreased protein expressions of CCR2, VLA4 and Mac-1 in THP-1 cells. Conclusion: TXL and Rb1 could protect HUVECs, reduce ox-LDL injury induced vascular endothelial cell adhesion and decrease relevant receptor expression in monocytes; therefore, inhibit injured monocytes adherence to vascular endothelial cells.
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Objective To evaluate the value of HE4 ,CA125 in the diagnosis of epithelial ovarian cancer (EOC) .Methods A to‐tal of 54 samples with epithelial ovarian cancer ,64 suspicious benign pelvic mass ,and 60 health controls were consecutively enrolled in this study .Results Good diagnostic performance in discriminating benign from EOC patients was obtained for CA125 and HE4 . Serum level of HE4 of the patients in the three group were 189 .94 pmol/L ,56 .74 pmol/L and 46 .36 pmol/L .Levels of CA125 were 89 .39 U/L ,45 .11 U/L and 34 .24 U/L .There were stastically significant differences between the groups(P<0 .05) .The ser‐um HE4 and CA125 levels of the patients with ovarian cancer at stage Ⅲ ,Ⅳ(HE4=236 .25 pmol/L ,CA125=206 .35 U/L) were significantly higher than the cases at stage Ⅰ ,Ⅱ(HE4=96 .36 pmol/L ,CA125=67 .8 U/L P<0 .05) .Benign pelvic mass as con‐trol ,the specificity (SP) was 86 .6% ,sensitivity (SN) was 82 .6% ,positive predictive value and negative predictive value of the ser‐um HE4 were 85 .71% ,71 .67% ,73 .84% ,and 84 .31% respectively .SN of CA125 was 86 .67% ,higher than that of HE4 (71 .67% ,P<0 .01) .The area under curves (AUC) of HE4 was 0 .87 higher than that of CA125 (0 .81) ,(P<0 .01) .Combing de‐tection of ovarian cancer was higher than that of HE4 and CA125 alone .SP was 95 .15% ,negative predictive value was 92 .13%(P<0 .01) .Conclusion Overall ,the level of serum CA125 and HE4 increase significantly ,which showes well diagnostic perform‐ance to EOC from benign diseases .The combined detection of CA125 and HE4 could improve the diagnostic power in cervical cancer prominently ,so it has great reference value in the diagnosis of epithelial ovarian cancer .
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Objective To observe the occurrence of hyperhomocysteinemia among patients suffering from cerebral infarction,and explore the possible risk factors.Methods Data of 144 cerebral infarction patients were collected from a tertiary hospital in Tianjin including gender,age,diabetes mellitus,hypertension,smoking,alcoholism,plasma homocysteine,HbAlc and biochemical criterion.Two groups were created as the hyperhomocysteinemia group and the control group based on serum homocysteine levels.Risk factor for hyperhomocysteinemia was analyzed by application of Logistic regression.Results Logistic regression analysis revealed that hyperhomocysteinemia was associated with gender,smoking,drinking and HbAlc levels in patients.Conclusions Male,smoking,drinking and diabetes mellitus were the possible risk factors in cerebral infarction patients.
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AIM: To explore the role of PI3K/Akt/nNOS in Zhouluotong extract resisting diabetic peripheral neuropathy.METHODS:The Schwann cells were divided into normal group ( D-glucose 25 mmol/L) , model group ( D-glucose 100 mmol/L) , Zhouluotong extract Z-6 +high glucose group, Zhouluotong +high glucose group, mecobalamine+high glucose group.The viability, nitric oxide content and the Ca2+-ATPase activity in Schwann cells were determined by Cell Counting Kit-8 , nitric oxide assay kit and Ca2+-ATPase assay kit, respectively.The apoptosis of Schwann cells were analyzed by flow cytometry.The expression of Bcl-2, Bcl-xL, Bax, Bak and caspase-3, and the phosphorylation levels of nNOS and Akt were determined by Western blotting.The signal pathway of PI3K/Akt was explored by dominant negative PI3K and Akt (δp85 and DN-Akt) transient transfection assay.RESULTS:Under high-glucose culture, the cell viability, nitric oxide content in culture supernatant, the expression of Bcl-2 and Bcl-xL, and the phosphorylation levels of Akt and nNOS in the Schwann cells were significantly increased.The cell apoptosis, the expression of Bax, Bak and caspase 3 in the Schwann cells were significantly decreased by Zhouluotong extract Z-6, compared with model group.In-creased nitric oxide content and the up-regulation of nNOS were observed.However, the effects of blocking PI3K/Akt, the upstream pathway of nNOS , by transfection with DN-δp85 on Akt phosphorylation in the Schwann cells was still unclear. CONCLUSION:Zhouluotong extract Z-6 changes the phosphorylation of nNOS, and the expression of anti-apoptotic fac-tors , caspase-3 and pro-apoptotic factors in Schwann cells under high-glucose culture, thus reducing apoptosis and elevating viability.The relationship to PI3K/Akt/nNOS pathway needs further investigation.
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AIM: To observe the different changes of neuroendocrine systems between the state of sport fatigue and poverty of movement. METHODS: 60 male Wistar rats were randomly divided into three groups: normal control group, sport fatigue model group and poverty of movement model group (20 rats in each group). The sport fatigue model was established by the method of combining basal diet and loaded swimming during 2 weeks, whereas the method of restricted activities was used to establish the poverty of movement model with total experimental time of 10 weeks. By the end of experiment, the climbing pole time was determined. The contents of hypothalamus thyrotropin releasing hormone (TRH), and serum norepinephrine (NE) and epinephrine (E) in rats with different treatments were determined by ELISA. In addition, the changes of hypothalamus corticotropin release hormone (CRH), pituitary adrenocorticotropic hormone (ACTH) and thyroid stimulating hormone (TSH), and serum corticosterone (CORT), triiodothyronine (T_3), tetraiodothyronine (T_4) were determined by radioimmunoassay to evaluate the functions of adrenergic nerve-adrenomedullin system, hypothalamo-pituitary-adrenal (HPA) axis and hypothalamo-pituitary-thyroid (HPT) axis. RESULTS: Compared to control group, the climbing pole time of the animals was obviously decreased in two model group. The adrenergic nerve-adrenomedullin system and HPA axis were inhibited in sport fatigue model rats, but HPT axis was unchanged. Interestingly, the HPA axis was hyperfunctional and HPT axis was inhibited in poverty of movement model rats. However, no change in the adrenergic nerve-adrenomedullin system was observed. CONCLUSION: Sport fatigue and poverty of movement all affect neuroendocrine system and lead to the adjustment mechanism imbalance, but the target and tendency are different.
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Objective:To investigate the vascular protective mechanism of different levels of herbs through detecting activity of HO-1/CO system in arterial tissues of rats with stagnancy of collateral-Qi in deficiency condition.Methods:Healthy male Wistar rats of cleanness level were randomly divided into control group,stagnancy of collateral-Qi in a deficiency condition group(SCQDC group),Renshen group,Shuangshen group,and Tongxinluo group.The ultrastructure of endothelium cells in the artery was observed.RT-PCR,Western blot,and immunohistochemical approaches were used to detect JNK,c-Jun mRNA and albumen expression in the arterial tissues.Meanwhile,the content of CO in the arterial tissue was also detected.Results:In SCQDC group,the counts of pinocytotic cells and microvili on the free surface of vascular endothelial cells were significantly decreased;most crests of mitochondria fused with membrane,some even disappeared;serious degranulation was observed in the rough surfaced endoplasmic reticulum.Immunohistochemistry showed that the HO-1 positive signals in the arterial histiocyte weakened significantly compared with the control group.The HO-1 expression was increased in the 3 treatment groups.Compared with control group,SCQDC group had decreased HO-1 mRNA and albumen expression(P
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AIM: To discuss the tongxinluo supermicropowder's interventional effect on aorta endodermis of rabbits which are fed with fatty forage plants.METHODS: Thirty-two healthy male New Zealand rabbits were divided into 4 groups randomly: control group,model group,atorvastatin treatment group,and tongxinluo treatment group.The control group was fed with common feedstuff,and all the other groups were fed with fatty feedstuff.The atorvastatin treatment group and the tongxinluo treatment group were given suspension of atorvastatin(3 g?kg-1?d-1) and tongxinluo supermicropowder(0.31 g?kg-1?d-1) by intragastric administration on the basis of fatty feedstuff.All the groups were fed with medicine for 6 weeks.At the end of 6 weeks,blood lipid levels were observed by enzymic method,the levels of blood serum NO and MDA and the activity of SOD were observed by chromatometry,the expression of nuclear factor-kappaB(NF-?B),intercellular adhesion molecule-1(ICAM-1) was detected,and VCAM-1mRNA expression in the aortic tissue was examined by reverse transcription polymerase chain reaction(RT-PCR).RESULTS: The levels of blood serum TC,TG,LDL-C,HDL-C,and MDA were increased significantly compared with the control group(P