HMGA2与胃癌上皮细胞间质转化的相关性及临床意义

探讨HMGA2、E-cadherin及N-cadherin在胃癌组织中差异性表达的临床意义及其与上皮细胞间质转化的关系.应用RT-PCR方法检测20例胃癌组织及对应的正常胃粘膜组织HMGA2 mRNA的表达,应用免疫组织化学法检测71例胃癌组织及癌旁组织中HMGA2、E-cadherin及N-cadherin的表达情况,与临床资料做统计学分析.HMGA2 mRNA在胃癌组织和癌旁组织中的表达存在显著性差异(P<0.01),在胃癌组织中HMGA2、E-cadherin及N-cadherin的阳性表达率分别为64.7%、29.6%和43.7%,HMGA2和E-cadherin的表达成负相关(P<0.05),和N-cadherin的表达成正相关(P<0.05),HMGA2表达和胃癌患者淋巴结转移及临床分期有显著相关性(P<0.05).HMGA2高表达的胃癌组织中有明显的上皮细胞间质转化表型,其表达与胃癌侵润转移有一定的相关性.     

Two new miR‐16 targets: caprin‐1 and HMGA1, proteins implicated in cell proliferation

Background information. miRNAs (microRNAs) are a class of non‐coding RNAs that inhibit gene expression by binding to recognition elements, mainly in the 3′ UTR (untranslated region) of mRNA. A single miRNA can target several hundred mRNAs, leading to a complex metabolic network. miR‐16 (miRNA‐16), located on chromosome 13q14, is involved in cell proliferation and apoptosis regulation; it may interfere with either oncogenic or tumour suppressor pathways, and is implicated in leukaemogenesis. These data prompted us to search for and validate novel targets of miR‐16. Results. In the present study, by using a combined bioinformatics and molecular approach, we identified two novel putative targets of miR‐16, caprin‐1 (cytoplasmic activation/proliferation‐associated protein‐1) and HMGA1 (high‐mobility group A1), and we also studied cyclin E which had been previously recognized as an miR‐16 target by bioinformatics database. Using luciferase activity assays, we demonstrated that miR‐16 interacts with the 3′ UTR of the three target mRNAs. We showed that miR‐16, in MCF‐7 and HeLa cell lines, down‐regulates the expression of caprin‐1, HMGA1a, HMGA1b and cyclin E at the protein level, and of cyclin E, HMGA1a and HMGA1b at the mRNA levels. Conclusions. Taken together, our data demonstrated that miR‐16 can negatively regulate two new targets, HMGA1 and caprin‐1, which are involved in cell proliferation. In addition, we also showed that the inhibition of cyclin E expression was due, at least in part, to a decrease in its mRNA stability.     

假基因HMGA1L2在甲状腺肿瘤中的表达

应用RT-PCR技术检测假基因HMGA1L2在50例良、恶性甲状腺病变中HMGA1L2 mRNA的表达。结果显示HMGA1L2 mRNA在12例结节性甲状腺肿、9例甲状腺腺瘤和15例甲状腺乳头状癌中的阳性表达率均为100%, 而在14例甲状腺滤泡癌中的阳性率为35.7%, 与前3者差异有显著性。该研究首次报告了假基因HMGA1L2 mRNA在良、恶性甲状腺病变中的表达, 并且提示其在甲状腺滤泡癌与腺瘤的鉴别诊断中具有潜在的价值。     

miR-98 acts as an inhibitor in chronic constriction injury-induced neuropathic pain via downregulation of high-mobility group AT-hook 2

Neuropathic pain, resulting from somatosensory nervous system dysfunction, remains a serious public health problem worldwide. microRNAs are involved in the physiological processes of neuropathic pain. However, the biological roles of miR-98 in neuropathic pain development have not been investigated. Therefore, in our current study, we focused on the effects of miR-98 in neuropathic pain. It was shown that miR-98 was significantly downregulated in chronic sciatic nerve injury (CCI) rat models. In addition, high mobility group A2 (HMGA2) was obviously upregulated in CCI rats. Overexpression of miR-98 inhibited neuropathic pain progression, including mechanical and thermal hyperalgesia. By a bioinformatics analysis, HMGA2 was predicted as a direct target of miR-98. The negative correlation between miR-98 and HMGA2 was validated in our present study. Furthermore, overexpression of miR-98 dramatically repressed HMGA2 protein and messenger RNA (mRNA) expression. Neuroinflammation participates in neural-immune interactions, which can contribute to the neuropathic pain development. Meanwhile, we found that inflammatory cytokine (interleukin [IL]-6, IL-1β, and COX-2) protein expression in rats infected with LV-miR-98 was greatly suppressed. Taking these results together, we concluded that miR-98 might depress neuropathic pain development through modulating HMGA2.     

FAD-linked presenilin-1 mutants impede translation regulation under ER stress

FAD mutations in presenilin-1 (PS1) cause attenuation of the induction of the endoplasmic reticulum (ER)-resident chaperone GRP78/BiP under ER stress, due to disturbed function of IRE1, the sensor for accumulation of unfolded protein in the ER lumen. PERK, an ER-resident transmembrane protein kinase, is also a sensor for the unfolded protein response (UPR), causing phosphorylation of eukaryotic initiation factor 2alpha (eIF2alpha) to inhibit translation initiation. Here, we report that the FAD mutant PS1 disturbs the UPR by attenuating both the activation of PERK and the phosphorylation of eIF2alpha. Consistent with the results of a disturbed UPR, inhibition of protein synthesis under ER stress was impaired in cells expressing PS1 mutants. These results suggest that mutant PS1 impedes general translational attenuation regulated by PERK and eIF2alpha, resulting in an increased load of newly synthesized proteins into the ER and subsequently increasing vulnerability to ER stress.     

Nrf2/HO-1/HMGB1信号通路在白血病细胞K562/A02化疗耐药中的机制研究

目的:通过观察高迁移率族蛋白1(HMGB1)、转录因子NF-E2相关因子2(Nrf2)及血红素加氧酶1(HO-1)基因沉默对白血病化疗耐药细胞(K562/A02细胞株)的影响,探讨该信号通路在白血病化疗耐药中的作用及其可能机制。方法:将HMGB1基因、Nrf2基因及HO-1基因的特异性干扰RNA分别转染阿霉素耐药细胞株K562/A02,荧光实时定量(RT-PCR)方法检测HMGB1、Nrf2及HO-1的mRNA表达水平,Western blot方法检测HMGB1、Nrf2及HO-1的蛋白表达水平,免疫荧光方法检测Nrf2的蛋白表达,并使用CCK-8方法检测转染前后K562/A02细胞株的细胞活性。结果:HMGB1基因、Nrf2基因或HO-1基因沉默的K562/A02细胞活性皆显著低于对照组及空白组(P0.05),化疗敏感性恢复。结论:HMGB1高表达导致了白血病细胞株K562/A02对阿霉素的化疗耐药,Nrf2/HO-1信号通路参与了HMGB1诱导的K562/A02细胞的化疗耐药,其表达上调可恢复K562/A02细胞对阿霉素的敏感性。     

The approximately 30-million-year-old ERVPb1 envelope gene is evolutionarily conserved among hominoids and Old World monkeys

Most human endogenous retroviruses (HERVs) are ancient and their genes are rendered nonfunctional by debilitating mutations. One exception is a recently discovered envelope gene located on chromosome 14. This envelope protein was also recently shown to be expressed in various human tissues and to mediate cell-cell fusion ex vivo. In this study, we demonstrate that this locus (designated ERVPb1) is preserved in Old World monkeys and that the reading frame is maintained. This is congruent with the entry of the HERV-P(b) group between 27 and 36 million years ago as suggested by long terminal repeat divergence. Although the coding capacity is generally lost in the HERV-IP supergroup, the analysis of nucleotide substitutions, lack of stop codons, and single-nucleotide polymorephisms strongly indicates a selective advantage of the ERVPb1 envelope genes during primate evolution. The purifying selection and tissue-specific expression of the human ERVPb1 envelope gene provide strong evidence of a beneficial role for the host.     

乳腺癌组织FOXP3、HMGA2、RASAL2的表达及与临床病理特征和预后的关系

摘要 目的：探讨乳腺癌组织叉头框蛋白P3（FOXP3）、高迁移率族蛋白A2（HMGA2）、大鼠肉瘤蛋白活化因子2（RASAL2）的表达及其与临床病理特征和预后的关系。方法：选取2014年6月至2015年6月期间我院收治的102例乳腺癌患者作为研究对象。检测乳腺癌组织以及癌旁组织中FOXP3、HMGA2、RASAL2表达,分析FOXP3、HMGA2、RASAL2表达与临床病理参数的相关性。Kaplan-Meier生存曲线分析不同FOXP3、HMGA2、RASAL2表达患者总生存率的差异。Cox比例风险回归模型分析乳腺癌患者预后的影响因素。结果：与癌旁组织相比,乳腺癌组织中FOXP3、HMGA2阳性表达率升高,而RASAL2阳性表达率降低（P<0.05）。FOXP3、HMGA2、RASAL2表达均与TNM分期和淋巴结转移相关（P<0.05）。FOXP3、HMGA2阳性患者的生存率明显低于FOXP3、HMGA2阴性患者,RASAL2阳性患者的生存率明显高于RASAL2阴性患者（P<0.05）。TNM分期Ⅲ期、FOXP3阳性表达、HMGA2阳性表达是影响乳腺癌患者预后的危险因素（P<0.05）,而RASAL2阳性表达是乳腺癌患者预后的保护因素（P<0.05）。结论：乳腺癌组织中FOXP3和HMGA2阳性表达率升高,而RASAL2阳性表达率降低,FOXP3和HMGA2阳性表达以及RASAL2阴性表达与乳腺癌患者TNM分期、淋巴结转移相关,并且是患者预后不良的影响因素。     

Role of high mobility group protein-1 (HMG1) in amyloid-beta homeostasis

In Alzheimer's disease (AD), fibrillar amyloid-beta (Abeta) peptides form senile plaques associated with activated microglia. Recent studies have indicated that microglial Abeta clearance is facilitated by several activators such as transforming growth factor-beta1 (TGF-beta1). The relationship between microglia and Abeta formation and deposition is still unclear. In the present study, high mobility group protein-1 (HMG1) inhibited the microglial uptake of Abeta (1-42) in the presence and absence of TGF-beta1. In addition, HMG1 bound to Abeta (1-42) and stabilized the oligomerization. In AD brains, protein levels of HMG1 were significantly increased in both the cytosolic and particulate fractions, and HMG1 and Abeta were colocalized in senile plaques associated with microglia. These results suggest that HMG1 may regulate the homeostasis of extracellular Abeta (1-42) and Abeta oligomerization.     

人乳头瘤病毒16E6基因通过增加HMGB1表达调节口腔癌CAL27细胞侵袭的实验研究

人乳头瘤病毒16型(Human papillomavirus type 16,HPV16)感染与口腔癌、宫颈癌的发病有关,HPV16 E6基因编码的蛋白是重要的癌蛋白,已经被证实能够通过增加高迁移率族蛋白B1(High mobility group box-B1,HMGB1)表达来促进宫颈癌细胞的侵袭,但是否能调控口腔癌细胞的侵袭仍未明确。为研究HPV16 E6基因通过增加HMGB1表达调节口腔癌CAL27细胞侵袭的作用,口腔癌CAL27细胞被分为对照组、空白质粒组、HPV16 E6质粒组、NC-si RNA组(短片断干扰RNA阴性对照组)、NC-si RNA+HPV16 E6质粒组、HMGB1-si RNA+HPV16E6质粒组,检测细胞中HPV16 E6及HMGB1的表达、细胞的侵袭数目、培养基中HMGB1的含量。结果显示,HPV16 E6质粒组细胞中HPV16 E6及HMGB1的表达量、培养基中HMBG1的含量、细胞的侵袭数目均高于对照组及空白质粒组(P<0.05);HMGB1-si RNA组细胞中HMGB1的表达量明显低于对照组及NC-si RNA组(P<0.05);NC-si RNA+HPV16 E6质粒组的细胞侵袭数目均明显高于NC-si RNA组(P<0.05),HMGB1-si RNA+HPV16 E6质粒组的细胞侵袭数目均明显低于NC-si RNA+HPV16 E6质粒组(P<0.05)。本研究提示,HPV16 E6基因能够促进口腔癌CAL27细胞的侵袭且这一作用与增加HMGB1表达有关。     

Antioxidant properties of UCP1 are evolutionarily conserved in mammals and buffer mitochondrial reactive oxygen species

Mitochondrial uncoupling reduces reactive oxygen species (ROS) production and appears to be important for cellular signaling/protection, making it a focus for the treatment of metabolic and age-related diseases. Whereas the physiological role of uncoupling protein 1 (UCP1) of brown adipose tissue is established for thermogenesis, the function of UCP1 in the reduction of ROS in cold-exposed animals is currently under debate. Here, we investigated the role of UCP1 in mitochondrial ROS handling in the Lesser hedgehog tenrec (Echinops telfairi), a unique protoendothermic Malagasy mammal with recently identified brown adipose tissue (BAT). We show that the reduction of ROS by UCP1 activity also occurs in BAT mitochondria of the tenrec, suggesting that the antioxidative role of UCP1 is an ancient mammalian trait. Our analysis shows that the quantity of UCP1 displays strong control over mitochondrial hydrogen peroxide release, whereas other factors, such as mild cold, nonshivering thermogenesis, oxidative capacity, and mitochondrial respiration, do not correlate. Furthermore, hydrogen peroxide release from recoupled BAT mitochondria was positively associated with mitochondrial membrane potential. These findings led to a model of UCP1 controlling mitochondrial ROS release and, presumably, being controlled by high membrane potential, as proposed in the canonical model of “mild uncoupling”. Our study further promotes a conserved role for UCP1 in the prevention of oxidative stress, which was presumably established during evolution before UCP1 was physiologically integrated into nonshivering thermogenesis.     

血清Hmga1、M-CSF及AFP与宫颈癌患者肿瘤病理特征及预后的关系

摘要 目的：探讨血清Hmga1、M-CSF、AFP与宫颈癌患者肿瘤病理特征及预后的关系。方法：选择2019年8月到2020年6月在本院妇产科诊治的120例宫颈癌患者作为宫颈癌组,同期选择宫颈良性病变患者120例作为良性组。检测两组患者的血清高迁移率族蛋白A1（Hmga1）、巨噬细胞集落刺激因子（M-CSF）、甲胎蛋白（AFP）含量,调查宫颈癌患者的肿瘤病理特征、预后情况并进行相关性分析。结果：宫颈癌组的血清Hmga1、M-CSF、AFP含量高于良性组（P<0.05）。在宫颈癌患者中,不同组织学分化、临床分期、淋巴结转移患者的血清Hmga1、M-CSF、AFP含量对比有差异（P<0.05）;随访到2022年1月1日,平均随访时间17.28±1.25个月,死亡18例,死亡率为15.0 %。Spearsman分析显示：患者预后死亡与Hmga1、M-CSF、AFP等存在相关性（P<0.05）。Cox比例风险模型分析显示Hmga1、M-CSF、AFP为影响患者预后死亡的重要因素（P<0.05）。结论：宫颈癌患者多存在血清Hmga1、M-CSF、AFP的高表达,且与患者的肿瘤病理特征存在相关性,宫颈癌患者的预后与血清Hmga1、M-CSF、AFP表达存在相关性,预测宫颈癌患者死亡具有很好的价值。     

单核细胞趋化蛋白-1、高迁移率族蛋白B1与溃疡性结肠炎患者肠道菌群变化的相关性

目的 探讨单核细胞趋化蛋白-1（MCP-1）、高迁移率族蛋白B1（HMGB1）与溃疡性结肠炎（UC）患者肠道菌群变化的相关性。方法 选取2016年9月‒2018年1月遂宁市中心医院收治的98例UC患者资料,根据Mayo评分系统将UC患者分为活动期组（n=50）和缓解期组（n=48）。选取同期进行体检的健康人50例作为对照组。比较各组间肠道菌群,血清MCP-1、HMGB1水平,并进行Pearson相关分析。结果 活动期组患者肠道乳杆菌、双歧杆菌含量[（5.34±0.87）、（5.81±0.83）CFU/g]显著低于缓解期组和对照组[（8.07±0.86）、（8.35±0.88）CFU/g;（8.13±0.91）、（8.46±0.95）CFU/g]（F=12.035,P0.05）。活动期组和缓解期组患者血清MCP-1、HMGB1水平[（267.42±23.51）、（21.35±2.26）ng/mL;（188.15±20.73）、（6.28±1.38）ng/mL]显著高于对照组[（106.38±15.92）、（2.13±0.41）ng/mL]（F=84.163,P<0.001;F=25.386,P<0.001）;活动期组患者血清MCP-1、HMGB1水平[（267.42±23.51）、（21.35±2.26）ng/mL]显著高于缓解期组[（188.15±20.73）、（6.28±1.38）ng/mL]（t=17.676、39.641,均P<0.05）。经过Pearson相关性分析,MCP-1、HMGB1与UC患者乳杆菌、双歧杆菌含量呈负相关（r=‒0.715、‒0.659,r=‒0.703、‒0.614,均P<0.001）,与大肠埃希菌、肠球菌、拟杆菌含量呈正相关（r=0.783、0.702,r=0.762、0.735,r=0.653、0.612,均P<0.001）。结论 MCP-1、HMGB1作为促炎因子可介导肠黏膜炎性反应,引起UC患者肠道菌群的紊乱。     

呼吸机相关性肺炎患者病原菌分布及血清HMGB1、PCT、CGRP水平变化的临床评价

探讨呼吸机相关性肺炎(VAP)患者感染病原菌分布情况以及血清高迁移率族蛋白B1(HMGB1)、降钙素原(PCT)、降钙素原基因相关肽(CGRP)水平的变化, 为该类患者的治疗提供参考。

选择2018年1月至2019年9月在我院接受治疗的机械通气患者137例, 其中符合VAP诊断标准的86例患者作为感染组, 其余51例作为未感染组; 选择同期在我院进行体检的60例健康者作为对照组。采用酶联免疫吸附法(ELISA)检测血清HMGB1、PCT、CGRP水平, 利用受试者工作曲线(ROC)预测血清HMGB1、PCT、CGRP对VAP的诊断价值。

137例机械通气治疗患者中有86例患者发生感染, 感染率为62.77%。86例感染患者样本共培养出95株致病菌, 其中革兰阴性菌(G^-)56株, 占58.95%;革兰阳性菌(G⁺)36株, 占37.89%;真菌3株, 占3.16%。感染组和未感染组患者血清HMGB1、PCT水平均高于对照组, CGRP水平低于对照组(均P < 0.05);感染组患者血清HMGB1、PCT水平均高于未感染组, CGRP水平低于未感染组(均P < 0.05)。ROC结果显示, 血清HMGB1、PCT、CGRP对VAP诊断的曲线下面积(AUC)分别为0.846、0.845、0.769, 最佳截断值分别为43.473μg/L、1.966μg/L、112.778 ng/L, 灵敏性分别为73.26%、65.12%、82.65%, 特异性分别为88.24%、94.12%、68.63%;三者联合诊断的AUC为0.932, 灵敏性和特异性分别为83.72%、94.12%。

VAP患者感染病原菌主要为革兰阴性菌。VAP患者血清HMGB1、PCT呈高表达, CGRP呈低表达, 三者对预测VAP均有一定效果, 其中三者联合诊断的效果最好, 具有一定的临床应用价值。

     

Evidence that high mobility group protein 17 is not phosphorylated in human colon carcinoma cells

The high mobility group proteins 14 and 17 were reported previously to be phosphorylated in murine and human tumor cell lines. Recently, it was suggested that subgroups of HMG-14, HMG-14a and 14b, but not HMG-17, were phosphorylated in situ in HeLa cells. In order to definitively determine whether HMG-17 is indeed phosphorylated or whether the protein previously identified as [³²P]HMG-17 was a subgroup of HMG-14, we have used the technique of electroblotting in conjunction with an immunochemical procedure utilizing anti-HMG-17 IgG. Our results indicate that HMG-17 was not phosphorylated in human colon carcinoma cell line HT-29 incubated for 18 h with ³²P_i, but that HMG-14a and HMG-14b were phosphorylated. In contrast, HMG-14a, -14b and -17 were phosphorylated in vitro in isolated nuclei incubated with [γ-³²P]ATP.     

急性肺损伤大鼠肺组织高迁移率族蛋白1的表达及其意义

目的动态观察高迁移率族蛋白1（HGMB1）在失血性休克复合内毒素注射致急性肺损伤（ALl）大鼠肺组织的表达情况,初步探讨HMGB1在ALI发病机制中的作用。方法采取失血性休克复合内毒素注射手段建立ALl大鼠动物模型,采用RT-PCR方法,检测肺组织HMGB1mRNA的表达情况。结果正常大鼠肺组织有少量HMGBlmRNA表达,遭受失血性休克复合内毒素注射打击后,HMGB1mRNA表达迅速升高,至ALI24h达最高峰,随后有所下降,ALl各组大鼠表达水平与正常对照组比较差异均有统计学意义（P〈0．01）。结论正常大鼠肺组织有一定水平HMGBlmRNA的表达,遭受失血性休克及内毒素注射打击后,HMGBlmRNA表达异常增高,可引起过度炎症反应,从而促进ALI的发生与发展。