Expression of genes for redox-dependent glutathione S-transferase isoforms GSTP1-1 and GSTA4-4 in tumor cell during the development doxorubicin resistance

Expression of genes for redox-dependent glutathione S-transferase isoforms GSTP1-1 and GSTA4-4 in tumor cells K562, MCF-7, and SKOV-3 was studied during the development of resistance to doxorubicin. It was found that the development of resistance was accompanied by predominant increase in the expression of hGSTP1 gene in MCF-7 cells, and hGSTA4 gene in resistant K562/DOX and SKVLB cells. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 143, No. 3, pp. 298–301, March, 2007     

Increased Expression of Glutathione by Estradiol, Tumor Necrosis Factor-Alpha, and Interleukin 1-Beta in Endometrial Stromal Cells

Problem  The intracellular antioxidant system, based on glutathione (GSH), plays a key role in endometrial detoxification reactions and has been proposed to be involved in the pathogenesis endometriosis. This study was designed to evaluate whether estradiol (E₂) and proinflammatory cytokines have any effects on expression of glutathione in endometrial stromal cells (ESCs).
Method of study  Glutathione levels were measured utilizing high-performance liquid chromatography following in vitro culture and treatment of ESCs with estradiol, tumor necrosis factor-alpha (TNF-α) and interleukin 1-beta (IL-1β).
Results  The GSH level in E₂ (10⁻⁸  m ) treatment group was significantly higher than in the control group at 48 h ( P  < 0.05). In vitro treatment of ESCs with TNF-α 10 ng/mL as well as E₂ (10⁻⁸  m ) plus TNF-α 10 ng/mL for 48 hr also led to a significant increase in GSH level ( P  < 0.05; P  < 0.05, respectively). Both IL-1β 10 ng/mL and E₂ (10⁻⁸  m ) plus IL-1β 10 ng/mL for 48 hr increased GSH level significantly ( P  < 0.05; P  < 0.05, respectively) as well.
Conclusions  These findings might suggest that increased production of estradiol and proinflammatory cytokines in the peritoneal cavity possibly leads to the establishment of endometriosis through increased level of GSH.     

GSTP1在前列腺癌组织中的表达及其甲基化检测

目的寻找前列腺癌患者血清中的生物标志物,为临床前列腺癌的早期无创性诊断提供依据。方法采用免疫组化SP染色法检测前列腺癌组织中GSTP1表达;运用甲基化特异性聚合酶链反应(MSP)方法检测前列腺癌患者癌组织和相应血清GSTP1基因启动子区5’端CpG岛甲基化程度。结果免疫组化结果:GSTP1在88例前列腺癌组仅有2例呈阳性反应且均为石蜡包埋标本;49例前列腺增生组均为阳性反应,其中38例呈现为强阳性反应。MSP检测结果:38例新鲜前列腺癌组织中,GSTP1基因高甲基化29例,此29例相应血清GSTP1基因高甲基化28例。对照组19例前列腺增生标本和对应的血清均没有检测到GSTP1基因甲基化。结论前列腺癌组织中GSTP1基因的表达与其启动子区甲基化程度呈负相关。前列腺癌患者的组织和血清中GSTP1甲基化检测结果相一致,检测血清中GSTP1的甲基化程度可反应癌组织中GSTP1基因的表达的情况。     

Expression of Laminin Isoforms,Receptors, and Binding Proteins Unique to Nucleus Pulposus Cells of Immature Intervertebral Disc

Intervertebral disc (IVD) disorders are believed to be related to aging-related cell loss and phenotypic changes, as well as biochemical and structural changes in the extracellular matrix of the nucleus pulposus (NP) region. Previously, we found that the laminin γ1 chain was more highly expressed in immature NP porcine tissues, in parallel with the expression pattern for a laminin receptor, integrin α6 subunit, as compared to adjacent anulus fibrosus region. This result suggests that cell-matrix interactions may be unique to the immature NP. However, the identity of laminin isoforms specific to immature or mature NP tissues, their associated receptors, and functional significance are still poorly understood. In this study, we evaluated the zonal-specific expression of the laminin chains, receptors (i.e., integrins), and other binding proteins in immature tissue and isolated cells of rat, porcine and human intervertebral disc. Our goal was to reveal features of cellular environment and cell-matrix interactions in the immature NP. Results from both immunohistochemical staining and flow cytometry analysis found that NP cells expressed higher levels of the laminin α5 chain, laminin receptors (integrin α3, α6, β4 subunit, and CD239), and related binding proteins (CD151), as compared to cells from adjacent anulus fibrosus. These differences suggest that laminin interactions with NP cells are distinct from that of the anulus fibrosus and that laminins may be important contributors to region-specific IVD biology. The revealed laminin isoforms, their receptors, and related binding proteins may be used as distinguishing features of these immature NP cells in the intervertebral disc.     

Effect of Nitric Oxide Donor,a Modulator of Tumor Drug Resistance,on Cell Death and p53 Protein Expression

Exogenous NO donor 3,3-bis-(nitroxymethyl)oxetane (NMO) was synthesized at the Institute for Problems of Chemical Physics (Russian Academy of Sciences). This compound was shown to inhibit cell death (apoptosis and necrosis) in cyclophosphamidesensitive and cyclophosphamide-resistant P388 murine tumor. p53 protein was expressed in both lines of tumor cells. NO donor NMO had little effect on p53 protein expression in cells of both stains. Our results suggest that the proapoptotic effect of NMO is mediated by the p53-independent molecular mechanisms.     

Analysis of the Expression of Regulator Genes in Kupffer Cells and Monocytes

Bulletin of Experimental Biology and Medicine - Differences in the gene expression profiles in resident macrophages (in particular, Kupffer cells) and monocytes were revealed. However, these...     

Expression of aquaporin-1, -2 and -4 in mice with a spontaneous mutation leading to hydronephrosis

This study investigates the expression of aquaporin-1, -2 and -4 in mice with a spontaneously-arising mutation that leads to hydronephrosis (ICR/Mlac-hydro mice). The mutant mice developed bilateral non-obstructive hydronephrosis without evidence of interstitial fibrosis or glomerulosclerosis. The mice had no abnormality in blood urea nitrogen or creatinine concentrations or in urine specific gravity. Despite the severity of the renal damage the mice grew and reproduced normally. Kidneys from the mutant mice had reduced expression of all three aquaporins compared with wild type mice. The reduction in aquaporin was proportional to the degree of hydronephrosis, but this change did not appear to be associated with disturbance of urinary function.     

Expression of Interferon Inducible Genes Following Hantaan Virus Infection as a Mechanism of Resistance in A549 Cells

Hantaan virus (HTN) is a causative agent of hemorrhagic fever with renal syndrome (HFRS). Little is known of its pathogenesis or the molecular mechanisms underlying resistance to HTN infection. In the present study, DNA microarray technology was used to monitor changes in mRNA levels after HTN infection, to elucidate resistance mechanisms to viral infection by understanding virus–host interactions. We found that several interferon (IFN)-inducible genes were up-regulated in host cells infected with HTN. According to previous available data, IFNs have been reported to be inhibitory, but their mode of action has not been yet clear. In this study, the 2,5-oligoadenylated synthetase (OAS) and M_x1 genes, not a double-stranded RNA-dependent protein kinase R (PKR), of the IFN response pathways are associated with antiviral activity during HTN infection. Furthermore, A549 cells treated with IFN- were protected against HTN infection. Taken together, these results confirmed that IFN plays a role in cellular defenses against HTN infection at an early stage of the infection and revealed the resistance mechanism for HTN infection.     

雄黄诱导MCF-7/ADM细胞凋亡及逆转其耐药性的研究

目的探讨雄黄诱导人乳腺癌MCF-7／ADM细胞凋亡及逆转其耐药性的调节机制。方法经MTT法探讨雄黄对人乳腺癌MCF-7／ADM细胞药物敏感性的影响;用透射电镜观察凋亡细胞形态改变;应用流式细胞术,探讨凋亡抑制基因bcl-2的编码蛋白Bcl-2的改变及凋亡百分率的改变;通过荧光分光光度法观察雄黄对细胞内化疗药物阿霉素积累的影响。结果雄黄对MCF-7／ADM的耐药性有明显的逆转作用,无毒剂量（15mg,L）及低毒剂量（25mg,L）雄黄作用后,逆转倍数分别为2．3倍及2．8倍;出现凋亡细胞,凋亡百分率由0．6％分别增至2．0％（P〈0．05）及3．4％（P〈0．01）;Bcl-2表达由90．2％分别降至63．6％（P〈0．01）及52．7％（P〈0．01）;MCF-7／ADM的细胞内阿霉素浓度明显增加（P〈0．01）。结论雄黄通过降低Bcl-2表达,促进细胞凋亡,增加耐药细胞内阿霉素积累量等耐药机制的调节,部分逆转了MCF-7／ADM细胞对阿霉素的耐药性。     

Resistance to Apoptosis and Expansion of Regulatory T Cells in Relation to the Detection of Circulating Tumor Cells in Patients with Metastatic Epithelial Cancer

Regulatory T cells may be crucial in the development of T cell tolerance to malignancies and contribute to immune dysfunctions. We investigated the percentage, activity, and onset of apoptosis of T cell subpopulations by multicolor flow cytometry in metastatic epithelial cancer patients compared to normal controls. Furthermore, a possible relationship between the presence of circulating tumor cells detected by immunocytochemistry and immune cell abnormalities was evaluated. Our study demonstrated a significantly elevated proportion of regulatory T cells in cancer patients (p < 0.001). In contrast to all other T cell subpopulations, regulatory T cells showed comparable Annexin V-binding characteristics in patients and normal controls. No relationship between the detection of circulating tumor cells and immune dysfunction was observed. These results indicate that cancer patients have a higher number of regulatory T cells with resistance to apoptotic stimuli partly responsible for immune dysfunctions as often observed in cancer patients.     

Expression and function of 4-1BB during CD4 versus CD8 T cell responses in vivo

4-1BBL(-/-) mice have a defect in recall CD8+ T cell responses to viruses, whereas CD4+ T cell responses to virus are unimpaired in these mice. In contrast, both CD4+ and CD8+ T cells respond to 4-1BB ligand (4-1BBL) in vitro. To clarify the role of 4-1BB/4-1BBL in CD4+ versus CD8+ T cell responses in vivo, we compared CD4 (OT-II) and CD8 (OT-I) TCR transgenic T cells responding to the same antigen in an in vivo adoptive transfer model in 4-1BBL(+/+) versus 4-1BBL(-/-) mice. During primary and secondary responses, expression of 4-1BB on in vivo-activated TCR transgenic T cells was earlier and more transient than previously observed in vitro, correlating with expression of the early activation antigen CD69 and preceding the transition to the CD44hi state. Although 4-1BB is expressed early in the primary response, there was no effect of 4-1BBL deficiency on initial CD8 T cell expansion and only a minor effect on initial CD4 T cell expansion. The major effect of 4-1BB/4-1BBL interaction is on the T cell recall response. This is due to effects of 4-1BBL on maintenance of T cell numbers at the end of the primary response with additional effects of 4-1BBL on secondary expansion of T cells.     

Expression of protease-activated receptor-1, -2, -3, and -4 in control and experimentally inflamed mouse bladder

Inflammation underlines all major bladder pathologies and represents a defense reaction to injury involving a mandatory participation of mast cells and sensory nerves. Mast cells are particularly frequent in close proximity to epithelial surfaces where they are strategically located in the bladder and release their mediators in response to inflammation. Tryptase is specifically produced by mast cells and modulates inflammation by activating protease-activated receptors (PARs). We recently found that PAR-4 mRNA is up-regulated in experimental bladder inflammation regardless of the initiating stimulus. Because it has been reported that PAR-1, PAR-2, and PAR-3 may also be involved in the processes of inflammation, we used immunohistochemistry to characterize the expression of all known PARs in normal, acute, and chronic inflamed mouse bladder. We found that all four PARs are present in the control mouse bladder, and follow a unique distribution. All four PARs are co-expressed in the urothelium, whereas PAR-1 and PAR-2 are predominant in the detrusor muscle, and PAR-4 is expressed in peripheral nerves and plexus cell bodies. The strong expression of PARs in the detrusor muscle indicates the need for studies on the role of these receptors in motility whereas the presence of PAR-4 in nerves may indicate its participation in neurogenic inflammation. In addition, PARs are differentially modulated during inflammation. PAR-1 and PAR-2 are down-regulated in acute inflammation whereas PAR-3 and PAR-4 are up-regulated. Bladder fibroblasts were found to present a clear demarcation in PAR expression secondary to acute and chronic inflammation. Our findings provide evidence of participation of PARs in the urinary system, provide a working model for mast cell tryptase signaling in the mouse bladder, and evoke testable hypotheses regarding the roles of PARs in bladder inflammation. It is timely to understand the role of tryptase signaling and PARs in the context of bladder biology.     

小鼠自身免疫性心肌炎发病过程中4-1BB/4-1BBL和IL-15的表达及意义

目的： 探讨在小鼠自身免疫性心肌炎发病过程中4-1BB/4-1BBL、IL-15的表达和变化,及其免疫学活性对心肌炎的影响。方法：将提纯的猪心肌肌球蛋白和完全弗氏佐剂等体积混合成乳浊液在1 d、8 d及30 d免疫具有遗传易感性的BALB/c小鼠,建立实验性自身免疫性心肌炎（EAM）模型。对照组小鼠仅用完全弗氏佐剂皮下注射。分别于初次免疫后21 d、80 d进行心肌炎症评分及血清肌钙蛋白I(cTnI)测定,免疫组化检测心肌淋巴细胞活化诱导受体配体（4-1BBL）的表达,ELISA法检测血清白细胞介素-15(IL-15)的浓度,RT-PCR技术检测4-1BB/4-1BBL和IL-15 mRNA在小鼠心肌组织中的表达。结果：在急性期21 d,EAM组小鼠心肌组织见不同程度的炎性细胞浸润和心肌细胞变性坏死、血清cTnI水平升高（P<0.05）;80 d EAM组小鼠心肌组织炎症减弱伴有纤维化出现、cTnI较前期降低;心肌4-1BBL和血清IL-15在EAM组中表达明显,在对照组中少量表达（P<0.05）;21 d EAM组小鼠心肌组织中4-1BB/4-1BBL和IL-15基因表达水平均高于对照组(P<0.01),且与心肌炎症呈明显的正相关,80 d时表达仍然升高(P<0.05)。结论：4-1BB/4-1BBL和IL-15在EAM小鼠发病过程中的表达上调,4-1BB/4-1BBL共刺激通路和IL-15可能协同参与了自身免疫性心肌炎的发生发展过程。     

Interaction of GSTM1, GSTT1, and GSTP1 Genotypes in Determination of Predisposition to Atopic Dermatitis

The frequency of individual genotypes GSTM1, GSTT1, and GSTP1 and haplotypes was determined in patients with atopic dermatitis and healthy children. The actual frequency of some haplotypes was far below the theoretical value. Some haplotypes were associated with predisposition and resistance to atopic dermatitis.