miR-204 inhibits proliferation of Hodgkin lymphoma cells by targeting Sirt1/p53 signaling pathway

AIM: To investigate the effect of microRNA-204 (miR-204) on the proliferation of Hodgkin lymphoma cells and the underlying mechanism. METHODS: The expression of miR-204 and Sirt1 mRNA in Hodgkin lymphoma tissues was detected by RT-qPCR. After transfection with miR-204 mimic, Sirt1 siRNA and miR-204 mimic+pcDNA3.1-Sirt1 into the L428 cells, the cell viability and BrdU incorporation were measured by CCK-8 assay and BrdU assay, respectively. The protein levels of Sirt1 and acetylated p53 (ac-p53) were determined by Western blot.The targeting relationship between miR-204 and Sirt1 was verified by double luciferase reporter assay. RESULTS: The low expression of miR-204 and the high mRNA expression of Sirt1 were found in the Hodgkin lymphoma tissues. Compared with control group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were significantly decreased after L428 cells were transfected with miR-204 mimic or Sirt1 siRNA (P<0.05). Compared with miR-204 mimic alone group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were increased after L428 cells were co-transfected with miR-204 mimic and pcDNA3.1-Sirt1 (P<0.05). The results of double luciferase reporter assay confiermed that Sirt1 was the target gene of miR-204. CONCLUSION: The inhibitory effect of miR-204 on the proliferation of L428 cells may be achieved by inhibiting the expression of Sirt1 and promoting the up-regulation of ac-p53.     

Osmolarity,cell volume and proliferation in nasopharyngeal carcinoma cells

AIM: To investigate the relationship between osmolarity, cell volume and cell proliferation in nasopharyngeal carcinoma cells. METHODS: MTT method was applied to detect the proliferation ability of the poorly-differentiated nasopharyngeal carcinoma cell (CNE-2Z) under various osmolarity conditions. The flow cytometry was used to analyse cell cycle distribution. Cell volume was obtained by the image analysis of living cells and cell viability was determined by the trypan blue assay. RESULTS: Cultivation of cells under the hypertonic conditions of 370 and 440 mOsmol/L increased cell volume by 8.7% and 27.8% and facilitated cell proliferation by 22.2% and 33.9%, respectively. However, hypotonic incubation of cells with osmolarity of 160 and 230 mOsmol/L decreased cell volume by 12.8% and 4.1% and inhibited cell proliferation by 34.0% and 15.6%, respectively. Cell volume was positively correlated with cell proliferation rate. Long-term cultivation of cells under anisotonic conditions did not significantly alter cell cycle distribution, but hypotonic cultivation decreased cell viability. CONCLUSION: Proliferation of nasopharyngeal carcinoma cells was closely correlated with the osmolarity of culture medium and cell volume. Hypotonic cultivation may inhibit cell proliferation by decreasing cell volume to facilitate cell death mechanisms.     

Effects of component of some Chinese herbs on proliferation of human umbilical vein endothelial cells in vitro

AIM: To observe the effects of some component of Chinese herbs for external use on proliferation of human umbilical vein endothelial cells (HUVEC) and investigate the mechanism of promoting tissue repair. METHODS: The method of MTT was used to examine the effects of Rg1, Rh1, perlolyrine, cinnamyl aldehyde, muscone, astragaluspolysaccharin (APS), velver antler polypeptide (VAP) and soluble extract of boswellia carterii birdw (BCB) on proliferation of HUVEC. RESULTS: APS did not promote proliferation of HUVEC at 9.75 mg/L-2.5 g/L; Rh1 promoted proliferation of HUVEC at 1.94 mg/L-0.5 g/L (P<0.05 or P＜0.01), and Rg1 inhibited proliferation of HUVEC at 31 mg/L (P<0.05); VAP promoted proliferation of HUVEC at 1 mg/L-0.5 g/L with optimal dose of 10 mg/L (P<0.01), Cinnamyl aldehyde promoted proliferation of HUVEC at 2 g/L(P＜0. 05); Muscone and soluble extract of BCB inhibited proliferation of HUVEC at 1 g/L, 0.5-2.5 kg/L(P＜0. 01), respectively; Perlolyrine inhibited proliferation of HUVEC at 0.125 g/L-0.5 g/L(P＜0. 01). CONCLUSION: The external herbs for supplementing Qi and warming Yang can promote HUVEC proliferation and improve angiogenesis during tissue repair. The external herbs for promoting blood circulation and accelerating capillary movement may have influence upon other stages of tissue repair.     

铜对体外仔猪软骨细胞增殖和细胞骨架的影响

体外分离、培养仔猪关节软骨细胞,在细胞培养液中分别添加铜0、7.8、15.6、31.2、62.5μmol/L。结果表明,软骨细胞在4种铜浓度中可存活并增殖,但随铜浓度的增加,其存活率、增殖率、3H-TdR掺入率有明显的差异,且能破坏软骨细胞骨架。培养液中添加铜31.2μmol/L,对软骨细胞的增殖作用最强,增殖率、3H-TdR掺入数显著高于对照组(P<0.01),软骨细胞形态及骨架均正常。表明31.2μmol/L铜浓度是促进体外软骨细胞增殖的最适浓度。     

断奶前补饲不同直/支链淀粉比开食料对羔羊瘤胃上皮发育的影响

为了研究断奶前补饲不同直/支链淀粉比开食料对羔羊瘤胃上皮发育的影响,选取24只体况良好、胎次一致、体重相近的10日龄羔羊(湖羊),将其随机分为3组,在饲喂相同母乳的基础上,分别补饲以纯木薯(CS)、玉米(MS)和豌豆淀粉(PS)(直/支链淀粉比分别约为0.11,0.27和0.44)为唯一淀粉来源的开食料。试验期间,每天4:00-19:00将羔羊抱入补饲栏补饲不同直/支链淀粉比的开食料,并且在6:30, 10:30和15:30将羔羊抱回母羊舍哺乳1 h。羔羊自由饮水,单栏饲喂,自由采食燕麦干草。56日龄时屠宰采样,采集瘤胃组织样品制作石蜡切片进行瘤胃乳头形态测定,采集瘤胃上皮样品提取RNA测定相关基因表达。结果表明:CS组羔羊瘤胃乳头长度和表面积显著(P<0.001)高于MS和PS组,CS和MS组羔羊瘤胃乳头宽度显著(P=0.001)高于PS组。对瘤胃上皮各层厚度统计显示,CS和PS组羔羊瘤胃上皮角质层厚度显著(P=0.001)高于MS组;CS和MS组羔羊瘤胃上皮颗粒层厚度显著高于(P<0.001)PS组;CS组羔羊瘤胃上皮棘基层厚度和总厚度显著(P<0.001)高于MS和PS组。qRT-PCR结果显示:不同直/支链淀粉比开食料显著影响羔羊瘤胃上皮CDK2和CDK6的mRNA表达量(CS>MS>PS, P<0.001);CS组羔羊瘤胃上皮细胞cyclin A和CDK4的mRNA表达量显著(P<0.05)高于PS组,但与MS组无显著差异;CS组羔羊瘤胃上皮cyclin D1的mRNA表达量显著(P=0.012)低于PS组,但与MS组无显著差异。CS组羔羊瘤胃上皮类胰岛素生长因子-1(insulin like growth factor,IGF-1)的mRNA表达量显著(P<0.001)高于MS和PS组,CS和MS组羔羊瘤胃上皮IGF-1R的mRNA表达量显著(P=0.001)高于PS组。上述结果表明:较高支链淀粉比开食料与较高直链淀粉比开食料相比,在瘤胃中较易降解生成挥发性脂肪酸,促进瘤胃上皮IGF-1及细胞周期蛋白mRNA的表达,进而有利于断奶前羔羊瘤胃上皮的发育。研究结果对制订羔羊营养方案具有重要指导意义。     

Clinical significance of KLF15 expression in human lung adenocarcinoma

AIM: To explore the clinical significance of Krüpple-like factor 15 (KLF15) protein expression in the patients with lung adenocarcinoma for exploring the therapeutic and prognositic biomarkers of lung cancer. METHODS: Four cases of lung adenocarcinoma tissues and matched adjacent tissues were collected from our hospital, and the expression of KLF15 protein in these tissues was analyzed by Western blot. At the same time, 72 cases of archived paraffin-embedded samples and clinical data of the patients with lung adenocarcinoma were also collected. The KLF15 protein expression in the archived paraffin-embedded lung adenocarcinoma samples was detected by immunohistochemical staining. The correlations between KLF15 protein expression and clinical characteristics of the patients including prognosis were also analyzed. In addition, the KLF15 protein was up-regulated in A549 cells, and then the effects of KLF15 protein on the viability of the cells were measured by CCK-8 assay. RESULTS: The protein expression of KLF15 in the 4 cases of lung adenocarcinoma tissues was significantly lower than that in matched paracancerous tissues. Fifty-three cases of lung adenocarcinoma specimens showed low expression or no expression of KLF15 protein in total 72 cases (73.6%). The 5-year survival rate of the patients with high expression of KLF15 protein in their specimens was higher than that of the patients with the low expression of KLF15 protein (P<0.01), and the expression of KLF15 protein was significantly correlated with the pathological staging (P<0.01) and T stage (P<0.01) of the patients with lung adenocarcinoma. Furthermore, the low expression of KLF15 protein was an important poor prognostic indicator of the patients. Up-regulation of KLF15 protein in the A549 cells significantly inhibited the growth of the cells. CONCLUSION: KLF15 inhibits the growth of lung adenocarcinoma cells. It could be used as a therapeutic target and a prognostic biomarker for the patients with lung adenocarcinoma.     

Micro-scale heterogeneity in urban forest soils affects fine root foraging by ornamental seedlings of Buddhist pine and Northeast yew

Urban soils are frequently characterized by a strong heterogeneity caused by intense anthropogenic activity and land use changes. Soil heterogeneity is commonly known to affect tree root development, but little has been detected concerning root foraging by ornamental trees in heterogeneous urban soils at micro-scale. In this study, Buddhist pine [Podocarpus macrophyllus (Thunb.) D. Don] and Northeast yew (Taxus cuspidata S. et Z.) were selected as ornamental tree species for a two-year study. In the first-year, seedlings were cultured under contrasting photoperiods to generate different morphologies. In the second year, seedlings were transplanted to pots filled with soils collected from an urban forest. Controlled-release fertilizers (N-P₂O₅-K₂O, 14-13-13) were evenly broadcasted to a half patch of the pot (heterogeneity) or to both halves (homogeneity) on the surface 5 cm beneath the pot-top at the rate of 0.135 g N seedling⁻¹. In the fertilized heterogeneous patch, larger Buddhist pine seedlings had greater dry weight, length, surface area, volume, number of tips, and morphological foraging-precision in fine roots. Compared to Northeast yew seedlings under natural photoperiod in the first year, those under the extended photoperiod had larger size, greater fine root biomass, and length but lower foraging-precision in the second year. N and P concentrations in second-year fine roots mainly increased with the availability of patches generated by fertilization for both species. In conclusion, the ability to forage for nutrients by ornamental tree seedlings in heterogeneous urban forest soils was species-specific. Buddhist pine seedlings had higher foraging precision in heterogeneous urban soils than Northeast yew seedlings due to their response to the extended photoperiod during culture.     

F-box domain is involved in regulating the proliferation of MCF-7 cells by FBXO39 protein

AIM: To investigate the effect of F-box domain on the regulation of MCF-7 cell proliferation by FBXO39 protein. METHODS: The effect of F-box domain on the localization of FBXO39 protein in the MCF-7 cells was investigated. MCF-7 cell cDNA library was used as the template resource. The full-length cDNA sequence of FBXO39 was amplified by PCR method and subcloned into eukaryotic expression vector pEGFP-C2. The pEGFP-FBXO39ΔF (F-box domain deletion mutation) plasmid was successfully constructed with the template resource of pEGFP-FBXO39 plasmid. The recombinant plasmids were transfected into the MCF-7 cells, and then the expression of FBXO39 and FBXO39ΔF were determined by Western blot. The cellular localization of FBXO39 and FBXO39ΔF were observed by confocal microscopy. The localization of endogenous FBXO39 in the MCF-7 cells was detected by immunofluorescence staining. In addition, MTT and EdU assays were used to measure the cell proliferation, flow cytometry was used to measure the cell cycle distribution, and immunohistochemical staining was used to observe the expression of FBXO39 in the breast cancer and para-carcinoma tissues. RESULTS: The eukaryotic expression vector pEGFP-FBXO39 and pEGFP-FBXO39ΔF were constructed successfully. F-box domain had no effect on the cell localization of FBXO39. FBXO39 promoted MCF-7 cell proliferation but FBXO39ΔF did not. FBXO39 was highly expressed in the breast cancer tissues. CONCLUSION: F-box domain had no effect on the cellular localization of FBXO39 protein. However, it plays an important role in the biological function of FBXO39. FBXO39 may be related to breast cancer tumorigenesis.