Osteopontin-deficient progenitor cells display enhanced differentiation to adipocytes

ObjectiveOsteopontin (OPN, Spp1) is a protein upregulated in white adipose tissue (WAT) of obese subjects. Deletion of OPN protects mice from high-fat diet-induced WAT inflammation and insulin resistance. However, the alterations mediated by loss of OPN in WAT before the obesogenic challenge have not yet been investigated. Therefore, we hypothesised that the lack of OPN might enhance the pro-adipogenic micro environment before obesity driven inflammation.MethodsOPN deficiency was tested in visceral (V) and subcutaneous (SC) WAT from WT and Spp1^−/− female mice. Gene expression for hypoxia, inflammation and adipogenesis was checked in WT vs. Spp1^−/− mice (n = 15). Adipocytes progenitor cells (APC) were isolated by fluorescence cell sorting and role of OPN deficiency in adipogenesis was investigated by cell images and RT-PCR.ResultsWe show that Spp1^−/− maintained normal body and fat-pad weights, although hypoxia and inflammation markers were significantly reduced. In contrast, expression of genes involved in adipogenesis was increased in WAT from Spp1^−/− mice. Strikingly, APC from Spp1^−/− were diminished but differentiated more efficiently to adipocytes than those from control mice.ConclusionsAPC from SC-WAT of lean OPN-deficient mice display an enhanced capacity for differentiating to adipocytes. These alterations may explain the healthy expansion of WAT in the OPN-deficient model which is associated with reduced inflammation and insulin resistance.     

乌苯美司联合XEC化疗方案治疗三阴性乳腺癌的临床研究

目的探讨乌苯美司联合XEC化疗方案治疗三阴性乳腺癌患者的临床疗效及其对外周血T淋巴细胞亚群及血清骨桥蛋白(OPN)水平变化的影响。方法选取河南科技大学第一附属医院2015年6月—2017年1月收治的三阴性乳腺癌患者83例,随机分成对照组(41例)和治疗组(42例)。对照组患者给予XEC化疗方案治疗,第1天静脉滴注注射用环磷酰胺500 mg/m~2,同时静脉滴注注射用盐酸表柔比星100 mg/m~2,第1~14天口服卡培他滨片1 g/m~2,疗程为21 d。治疗组在对照组的基础上口服乌苯美司片,30 mg/次,1次/d,疗程为21 d。两组均持续治疗4个疗程。观察两组患者临床疗效,比较治疗前后两组患者SF-36评分、外周血T淋巴细胞亚群表达水平和OPN水平。结果治疗后,对照组和治疗组临床总有效率分别为75.61%、95.24%,两组比较差异具有统计学意义(P0.05)。治疗后,两组躯体疼痛、躯体功能、睡眠质量、生命活力、生理职能、心理健康等生活质量评分均显著升高,同组治疗前后比较差异具有统计学意义(P0.05);且治疗后治疗组生活质量评分显著高于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组患者CD~(3+)、CD~(4+)、CD~(4+)/CD~(8+)水平均显著升高,CD~(8+)水平显著下降,同组治疗前后比较差异具有统计学意义(P0.05);且治疗组T淋巴细胞亚群水平显著优于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组血清OPN水平显著下降,同组治疗前后比较差异具有统计学意义(P0.05);且治疗组血清OPN水平显著低于对照组,两组比较差异具有统计学意义(P0.05)。结论乌苯美司联合XEC化疗方案治疗三阴性乳腺癌患者效果显著,可有效改善患者外周血T淋巴细胞亚群和血清OPN水平,且安全性较高,具有一定的临床推广应用价值。     

Cloning of hamster osteopontin and expression distribution in normal tissues and experimentally induced oral squamous-cell carcinoma

Osteopontin (OPN) is a non-collagenous extracellular matrix (ECM) protein expressed and secreted by several human cancers. This study investigated the expression pattern of OPN during development of oral squamous-cell carcinoma by using 7,12-dimethylbenz[a]anthracene (DMBA)-induced squamous-cell carcinomas in buccal pouch of syrian golden hamsters. We first identified the hamster OPN cDNA sequence by screening of a hamster calvariae cDNA library with a rat OPN cDNA probe. The resulting 1,449 bp of hamster OPN cDNA led to a deduced protein sequence of 305 amino acids containing several putative binding sites to integrins, CD44 receptors, calcium ions and hydroxyapatite, as well as multiple sites for phosphorylation, glycosylation and sulphation. Hamster OPN cDNA was then used as a probe to analyze the expression of OPN mRNA by Northern blot and in situ hybridization analyses of normal and malignant tissues. OPN mRNA was detected in several non-mineralized tissues as well as in mineralized tissues, but was not present in normal hamster buccal epithelium. DMBA-treated hamster buccal pouches expressed OPN mRNA as early as 4 weeks and displayed the highest level of expression at 15 weeks. The specimens treated with DMBA for 15 weeks exhibited histological features of squamous-cell carcinoma, presented microcrystalline deposits and showed OPN expression associated with malignant epithelium and tumor-associated macrophages. To summarize, our results suggest that buccal-pouch carcinogenesis of Syrian golden hamster may constitute an excellent experimental model to study the mechanisms by which OPN is associated with oral cancer pathogenesis, and to validate OPN-based therapeutic approaches to ameliorate oral cancer progression and metastasis.     

骨桥蛋白对骨髓间充质干细胞核力学特性的影响及相关分子机制

目的研究骨桥蛋白(osteopontin,OPN)对骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMSCs)核力学特性的影响及相关分子机制。方法采用Transwell法分析BMSCs迁移能力。利用原子力显微镜(atomic force microscope,AFM)检测细胞核弹性模量,分析OPN作用下BMSCs细胞核硬度的变化。通过Western blot技术检测OPN作用对黏着斑激酶(focal adhesion kinase,FAK)和胞外信号调节激酶1/2(extracellular signal-regulated kinase 1/2,ERK 1/2)的影响,并利用FAK或ERK1/2抑制剂考察FAK-ERK1/2信号通路在OPN影响BMSCs核力学特性中的作用。通过RT-PCR和Western blot技术检测OPN作用下核纤层蛋白Lamin A/C的表达变化。结果 OPN处理组细胞核弹性模量与对照组相比明显降低。OPN作用显著上调FAK、ERK1/2磷酸化水平,加入FAK或ERK1/2抑制剂在一定程度上回救OPN降低的细胞核弹性模量,并显著抑制BMSCs迁移。OPN处理BMSCs后Lamin A/C mRNA和蛋白水平的表达出现下调,但FAK或ERK1/2抑制剂能够抑制OPN诱导的Lamin A/C表达下调。结论 OPN可能通过FAK-ERK1/2信号通路下调BMSCs核骨架蛋白Lamin A/C表达,降低细胞核硬度,促进BMSCs迁移。该研究结果为深入认识OPN调控BMSCs迁移行为的机制及其临床应用提供了实验依据。     

Correlation between calcification and bone sialoprotein and osteopontin in papillary thyroid carcinoma

The correlation between calcification and papillary thyroid carcinoma has received increasing attention. We investigated the ability of bone sialoprotein (BSP) and osteopontin (OPN) protein levels to diagnose papillary thyroid carcinoma (PTC), and explored the correlation between BSP and OPN protein levels and calcification in PTC. Archival PTC specimens from patients with PTC with calcification and lateral cervical lymph node metastasis (LNM) were included in this retrospective immunohistochemical study. The protein levels of BSP and OPN were analysed immunohistochemically using routinely prepared tissue sections. PTC specimens from 66 patients with PTC were reviewed retrospectively (25 patients with histological calcification seen in paraffin sections, 41 patients without calcification; 35 patients with lateral cervical LNM, 31 patients without LNM). The percentage of samples that had cells that demonstrated positive protein staining differed significantly between PTC specimens, benign thyroid nodules, and adjacent normal follicular epithelium (BSP: 87.88%, 55.00%, and 42.50%, respectively; OPN: 83.33%, 70.00% and 50.00%, respectively). There was a significant difference in the immunohistochemical score (IHS) for BSP and OPN protein staining between PTC specimens with and without calcification (P < 0.05). The level of BSP protein staining was found to be significantly correlated with the level of OPN protein staining in PTC specimens. We conclude that the strong correlation between BSP and OPN and PTC suggests a role for BSP and OPN in calcification and tumor progression of PTC. BSP and OPN might be useful tumour markers for the diagnosis of PTC with limited value, because both of them had low specificity.     

消癌平胶囊联合顺铂和替加氟治疗晚期食管癌的临床研究

目的探讨消癌平胶囊联合顺铂和替加氟注射液治疗食管癌的临床疗效。方法选取2016年4月—2018年4月在重庆市开州区人民医院治疗的食管癌患者86例,根据用药方案的差别分为对照组(43例)和治疗组(43例)。对照组静脉滴注顺铂注射液,80 mg/m~2,1次/d,连续3周,休息1周,每4周为1个周期,同时静脉滴注替加氟注射液,15～20 mg/kg溶于生理盐水500 mL中,1次/d。治疗组在对照组基础上口服消癌平胶囊,4粒/次,3次/d。两组患者均治疗3个周期。观察两组患者临床疗效,同时比较治疗前后两组患者癌胚抗原(CEA)、鳞状上皮细胞癌抗原(SCC-Ag)、细胞角蛋白19的可溶性片段(CYFRA21-1)、糖抗原199(CA199)、血管内皮生长因子(VEGF)、基质金属蛋白酶-9(MMP-9)、骨桥蛋白(OPN)、可溶性白细胞介素-2受体(s IL-2R)及QLQ-C30和KPS评分。结果治疗后,对照组客观缓解率和临床获益率分别为41.86%、72.09%,均分别显著低于治疗组的69.77%、86.05%,两组比较差异均具有统计学意义(P0.05)。治疗后,两组患者血清CEA、SCC-Ag、CYFRA21-1、CA199、VEGF、MMP-9、OPN、sIL-2R水平均显著降低(P0.05),且治疗组患者这些因子水平明显低于对照组(P0.05)。治疗后,两组QLQ-C30量表和KPS评分均明显升高(P0.05),且治疗组患者QLQ-C30和KPS评分明显高于对照组(P0.05)。结论消癌平胶囊联合顺铂与替加氟治疗食管癌可有效改善患者症状,降低机体肿瘤标志物水平和侵袭细胞因子水平,有利于提高患者生活质量。     

OPN4 belongs to the photosensitive system of the human skin

The human skin has previously been described to be affected by light; however, the underlying mechanism remains unknown. OPN4 (melanopsin) expression was first identified in the skin of amphibians; however, whether it is also expressed and functioned in the human skin has not yet been identified. Here, we show that OPN4 was expressed in the human skin tissue and cultures of isolated keratinocytes, melanocytes and fibroblasts. Additionally, Ca²⁺ influx in vitro and ex vivo and phosphorylation of extracellular signal‐regulated kinases 1/2 in human fibroblasts were observed by stimulation of blue light irradiation. Notably, our findings showed that this Ca²⁺ influx and phosphorylation of extracellular signal‐regulated kinases 1/2 are promoted in an intensity‐dependent manner, indicating that the light signal is converted to an intracellular signal via OPN4 in the human skin. Overall, in this study we showed that the human skin functions as a photoreceptor by demonstrating that in human skin, the photoreceptive protein was expressed, and photoreception was conducted via photoreceptive protein.     

骨桥蛋白在卵巢癌中的表达及其与不同临床病理特征关系的Meta分析

目的:评价有关骨桥蛋白(OPN)在卵巢癌组织中表达的情况及与卵巢癌不同临床病理特征的关系。方法:系统检索NCBI PubMed、EMBASE、维普、万方数据库中截止2013年11月符合纳入标准的国内外公开发表的有关OPN在卵巢癌组织中表达的文献7篇,通过固定效应模型综合分析合并效应量,采用RevMan 5.1统计软件进行分析。结果:Meta分析显示,OPN在卵巢癌组和正常卵巢组表达差异有统计学意义[OR=20.24,95%CI(9.66,42.43),P<0.000 01]。OPN在卵巢癌组和良性卵巢组表达差异有统计学意义[OR=16.58,95%CI(9.60,28.64),P<0.000 01]。OPN在卵巢癌高分化组和中低分化组表达的差异有统计学意义[OR=0.16,95%CI(0.08,0.32),P<0.000 01]。OPN在卵巢癌临床Ⅰ-Ⅱ期组和临床Ⅲ-Ⅳ期组表达的差异有统计学意义[OR=0.16,95%CI(0.09,0.29),P<0.000 01]。OPN在卵巢癌淋巴结转移组和淋巴结未转移组表达的差异有统计学意义[OR=4.20,95%CI(2.24,7.88),P<0.000 01]。结论:OPN高表达与卵巢癌及其不同临床病理特征有显著相关性。但受纳入文献质量影响,还需进一步用国内外高质量的研究来证实。