Cervical cancer with transferrin receptor has a poor prognosis and is associated with immune infiltration, according to a comprehensive bioinformatics study

Background:The molecular mechanism underlying the involvement of the Transferrin receptor(TFRC)in cervical cancer remains poorly understood.This study aims to elucidate the role of TFRC in cervical cancer by analyzing data from The Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases.Methods:TFRC protein expression was obtained from Human Protein Altas(HPA).All datas were collected from TCGA and GTEx.In this study,we analyzed the expression of TFRC in cervical cancer and its clinical significance.Through Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene set enrichment analyses(GSEA),investigated the related molecular pathways of TFRC.The relationship between TFRC and immune infiltration was then examined.The prognosis of different immune cell subsets was then analyzed after dividing cervical cancer patients into high and low expression of TFRC groups.Results:TFRC is highly expressed in various tumor tissues compared to control normal tissues,including cervical cancer.An increased expression of TFRC was associated with higher Tumor(T)and Node(N)stage,as well as a higher clinical stage.Kaplan–Meier(KM)survival analysis investigated that higher TFRC expression patients have a poor overall survival(OS),disease specific survival(DSS)and progress free interval(PFI).Both KEGG and GSEA enriched signaling pathway by high TFRC and low TFRC groups.There was a significant negative linear correlation between TFRC expression and immune infiltration.TFRC affects the prognosis of cervical cancer patients through immune pathway.Conclusions:Cervical cancer patients with TFRC expression may have a worse prognosis.

Transferrin receptor and ferritin-H are developmentally regulated in oligodendrocyte lineage cells

Iron is an essential trophic element that is required for cell viability and differentiation, especially in oligodendrocytes, which consume relatively high rates of energy to produce myelin. Multiple iron metabolism proteins are expressed in the brain including transferrin receptor and ferritin-H. However, it is still unknown whether they are developmentally regulated in oligodendrocyte lineage cells for myelination. Here, using an in vitro cultured differentiation model of oligodendrocytes, we found that both transferrin receptor and ferritin-H are significantly upregulated during oligodendrocyte maturation, implying the essential role of iron in the development of oligodendrocytes. Additional different doses of Fe3＋ in the cultured medium did not affect oligodendrocyte precursor cell maturation or ferritin-H expression but decreased the expression of the transferrin receptor. These results indicate that upregulation of both transferrin receptor and ferritin-H contributes to maturation and myelination of oligodendrocyte precursor cells.

Study on The Method of Quantitative Analysis of Serum Ferritin and Soluble Transferrin Receptor with Protein Microarray Technology

Objective To establish and evaluate a protein serum ferritin （SF） and soluble transferrin receptor microarray method for combined measurement of （sTfR）. Methods Microarrayer was used to print both anti-SF antibodies I and anti-sTfR antibodies I on each protein microarray. Anti-SF antibodies II and anti-sTfR antibodies II were used as detection antibodies and goat antibodies coupled to Cy3 were used as antibodies Ill. The detection conditions of the quantitative analysis method for simultaneous measurement of SF and sTfR with protein microarray were optimized and evaluated. The protein microarray was compared with commercially available traditional tests with 26 serum samples. Results By comparison experiment, mouse monoclonal antibodies were chosen as the probes and contact printing was chosen as the printing method. The concentrations of SF and sTfR probes were 0.5 mg/mL and 0.5 mg/mL respectively, while those of SF and sTfR detection antibodies were 5 μg/mL and 0.36 μg/mL respectively. Intra- and inter-assay variability was between 3.26% and 18.38% for all tests. The regression coefficients comparing protein microarray with traditional test assays were better than 0.81 for SF and sTfR. Conclusion The present study has established a protein microarray method for combined measurement of SF and sTfR.

Preparation and Identification of scFv and bsFv against Transferrin Receptor

To obtain single chain variable fragment （scFv） and bivalent single chain variable fragment （bsFv） against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy （VH） and FR4 of variable light （VL）, respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.

Selection for Anti-transferrin Receptor Bispecific T-cell Engager in Different Molecular Formats

Selecting an ideal molecular format from diverse structures is a major challenge in developing a bispecific antibody(BsAb).To choose an ideal format of anti-CD3 x anti-transferrin receptor(TfR)bispecific antibodies for clinical application,we constructed TfR bispecific T-cell engager(BiTE)in two extensively applied formats,including single-chain tandem singlechain variable fragments(scFvs)and double-chain diabodies,and evaluated their functional characterizations in vitro.Results demonstrated that TfR-BiTE in both formats directed potent killing of TfR+HepG2 cells.However,compared to two・chain diabodies,scFvs were more efficient in antigen binding and TfR target killing.Furthermore,different domain orders in scFvs would also be evaluated because single-TfR-CD3-His was preferable to single-CD3-TfR-His in immunotherapeutic strategies.Thus,the single-chain tandem TfR-CD3 format was favored for further investigation in cancer therapy.

Effect of high flux hemodialysis on renal anemia and soluble transferrin receptor in hemodialysis patients

Objective: To investigate the effect of high throughput hemodialysis on soluble transferrin receptor in hemodialysis patients and the improvement of renal anemia. Methods: 132 patients receiving maintenance hemodialysis in our hospital from July 2017 to July 2019 were selected and divided into control group and observation group according to the random number table method, with 66 cases each. The observation group was treated with high-flux hemodialysis, while the control group was treated with low-flux hemodialysis for 6 months. Compare two groups before and after treatment serum beta 2 microglobulin (beta 2 - MG), serum creatinine (Scr), blood urea nitrogen (BUN) level, anemia related index [red blood cells deposited (HCT), hemoglobin (Hb), reticulocyte percentage (Ret%)], iron metabolism index [serum ferritin (SF), transferrin saturation (TSAT)、Hepcidin(Hepc)], soluble transferrin receptor (sTfR) levels and adverse reactions. Results: the levels of 2-MG, Scr and BUN in the two groups before treatment were compared (P>0.05). After treatment, Scr and BUN levels in the two groups were significantly decreased (P<0.05), but were compared between the two groups (P>0.05). The level of 2-MG in the observation group was lower than that in the control group (P<0.05). Before treatment, sTfR, Hb, HCT level and Ret% of the two groups were compared(P>0.05). After treatment, Hb and HCT levels in the observation group were higher than those in the control group, while Ret% were lower than those in the control group, (P<0.05). Before treatment, the levels of ST、TAST、sTfR and Hepc in the two groups were compared (P>0.05). After treatment, the level of ST and TAST in the observation group was higher than that in the control group, The levels of sTfR and Hepc were lower than the control group (P<0.05). The overall incidence of adverse reactions in the observation group (8.93%) was lower than that in the control group (10.14%), with no significant difference (P>0.05). Conclusion: The high-throughput hemodialysis department significantly improved renal anemia in hemodialysis patients, reduced serum sTfR level, and had fewer adverse reactions and higher safety.

Construction of single chain Fv antibody against transferrin receptor and its protein fusion with alkaline phosphatase

AIM: To construct fusion protein of a single-chain antibody(scFv) against transferrin receptor (TfR) with alkalinephosphatase (AP).METHODS: The VH-linker-VL, namely scFv gene, wasprepared by amplifying the VH and VL genes from plasmid pGEM-T-VH and pGEM-T-VL with splicing overlap extension polymerase chain reaction (SOE PCR). After the ScFv gene was modified by SfiⅠ and NotⅠ, it was subcloned into the secretory expression vector pUC19/119, and then was transformed into E. coli TG1. The positive colonies were screened by colony PCR and their expressions were induced by IPTG. ScFv gene was gained by digesting ScFv expression vector pUC19/119 with Sfi I and NotⅠ restriction enzymes, then subcloned into expression vector pDAP2, followed by transformation in E. coli TG1. The positive colonies were selected by bacterial colony PCR. The expression of fusion protein (scFv-AP) was induced by IPTG. Its activity was detected by enzyme immunoassay. The molecular weights of scFv and scFv-AP were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).RESULTS: The product of SOE PCR formed a band of 700 bp in agarose gel electrophoresis. SDS-PAGE demonstrated the molecular weight of scFv was 27 ku. Immunofluorescent assay (IFA) demonstrated its reactivity with TfR. The molecular weight of scFv-AP was 75 ku. Enzyme immunoassay showed that scFv-AP could specifically bind to human TfR and play AP activity.CONCLUSION: We have successfully prepared the antihuman TfR scFv and constructed the fusion protein of scFv and AP. It is promising for immunological experiments.

Serum Transferrin Receptors in Children with Hypochromic Microcytic Anaemia

Hypochromic microcytic anaemia includes iron deficiency, anaemia of chronic disorders, beta thalassemia trait and sideroblastic anaemia. To rule out the cause of hypochromic microcytic anaemia is a diagnostic difficulty. The conventional laboratory tests used for diagnosis have few disadvantages. Serum transferrin receptor (sTfR) is the most reliable method for assessment of body iron. Eighty four children were included in this study. They were further divided into four groups: iron deficiency anaemia (IDA), anaemia of chronic disorders (ACD), beta thalassemia trait (β TT) and controls. Children withIDAand ACD were diagnosed on the basis of history and serum iron profile. Subjects with β TT had HbA2 > 3.5%. sTfR were performed on all subjects. Level of sTfR in patients withIDAwas 5.79 μg/ml ± 1.3 μg/ml. In patients with anaemia of chronic disorders (ACD), β thalassemia trait and controls mean sTfR were 2.18 μg/ml ± 0.6 μg/ml, 2.1μg/ml ± 0.5 μg/ml and 2.0 μg/ml ± 0.5 μg/ml respectively. These results show level of sTfR was raised in IDA when compared with controls or ACD and β TT (p

Role of transferrin receptor in hepatitis C viral infection

Hepatitis C virus(HCV) is the main pathogen causing chronic hepatitis and primary liver cancer. Various viral proteins and host cell molecules are involved in the HCV cell entry, but the mechanism of infection has not been completely elucidated. The transferrin receptor can act as a receptor for many viruses during cell entry. The transferrin receptor is not only closely related to HCV-induced iron metabolism disorders but also mediates the fusion of HCV with the host cell membrane as a specific receptor for CD81-dependent viral adhesion.

Transferrin receptor and Fc α/μ receptor may not be the major IgA_1 receptor on human mesangial cells

IgA nephropathy (IgAN) is the most common form of primary glomerulonephritis. 1 The histopathology of IgAN is characterized by abundance of mesangial matrix and proliferation of mesangial cells. IgA_1 deposition in the mesangium plays an important role in the inflammatory process in this disease.

Eph receptor A4 regulates motor neuron ferroptosis in spinal cord ischemia/reperfusion injury in rats

Previous studies have shown that the receptor tyrosine kinase Eph receptor A4(EphA4) is abundantly expressed in the nervous system. The EphA4 signaling pathway plays an important role in regulating motor neuron ferroptosis in motor neuron disease. To investigate whether EphA4 signaling is involved in ferroptosis in spinal cord ischemia/reperfusion injury, in this study we established a rat model of spinal cord ischemia/reperfusion injury by clamping the left carotid artery and the left subclavian artery. We found that spinal cord ischemia/reperfusion injury increased EphA4 expression in the neurons of anterior horn, markedly worsened ferroptosis-related indicators, substantially increased the number of mitochondria exhibiting features consistent with ferroptosis, promoted deterioration of motor nerve function, increased the permeability of the blood-spinal cord barrier, and increased the rate of motor neuron death. Inhibition of EphA4 largely rescued these effects. However, intrathecal administration of the ferroptosis inducer Erastin counteracted the beneficial effects conferred by treatment with the EphA4 inhibitor. Mass spectrometry and a PubMed search were performed to identify proteins that interact with EphA4, with the most notable being Beclin1 and Erk1/2. Our results showed that inhibition of EphA4 expression reduced binding to Beclin1, markedly reduced p-Beclin1, and reduced Beclin1-XCT complex formation. Inhibition of EphA4 also reduced binding to p-Erk1/2 and markedly decreased the expression of c-Myc, transferrin receptor 1, and p-Erk1/2. Additionally, we observed co-localization of EphA4 and p-Beclin1 and of EphA4 and p-ERK1/2 in neurons in the anterior horn. In conclusion, EphA4 participates in regulating ferroptosis of spinal motor neurons in the anterior horn in spinal cord ischemia/reperfusion injury by promoting formation of the Beclin1-XCT complex and activating the Erk1/2/c-Myc/transferrin receptor 1 axis.

Actively priming autophagic cell death with novel transferrin receptor-targeted nanomedicine for synergistic chemotherapy against breast cancer

Recently, considerable attention in the field of cancer therapy has been focused on the mammalian rapamycin target(m TOR), inhibition of which could result in autophagic cell death(ACD). Though novel combination chemotherapy of autophagy inducers with chemotherapeutic agents is extensively investigated, nanomedicine-based combination therapy for ACD remains in infancy. In attempt to actively trigger ACD for synergistic chemotherapy, here we incorporated autophagy inducer rapamycin(RAP) into 7 pep-modified PEG-DSPE polymer micelles(7 pep-M-RAP) to specifically target and efficiently priming ACD of MCF-7 human breast cancer cells with high expression of transferrin receptor(Tf R). Cytotoxic paclitaxel(PTX)-loaded micelle(7 pep-M-PTX) was regarded as chemotherapeutic drug model. We discovered that with superior intracellular uptake in vitro and more tumor accumulation of micelles in vivo, 7 pep-M-RAP exhibited excellent autophagy induction and synergistic antitumor efficacy with 7 pep-M-PTX. Mechanism study further revealed that 7 pep-M-RAP and 7 pep-MPTX used in combination provided enhanced efficacy through induction of both apoptosis-and mitochondria-associated autophagic cell death. Together, our findings suggested that the targeted excess autophagy may provide a rational strategy to improve therapeutic outcome of breast cancer, and simultaneous induction of ACD and apoptosis may be a promising anticancer modality.

Ultraviolet-B induced expression of hypoxia-inducible factor 1α, transferrin receptor through EGFR/PI3K/AKT/DEC1 pathway

The aim of this research was to explore the effects and signaling pathway of ultraviolet-B(UVB)irradiation on the expression of hypoxia-inducible factor 1a(HIF-1α)and transferrin receptor(TfR).HIF-1α protein was measured by Western blot method.Expressions of epidermal growth factor receptor(EGFR),phosphor-EGF-R and TfR after UVB irradiation were determined with flow cytometry.After UVB irradiation,mRNA levels of HIF-1α and TfR were detected by real time-PCR.Results showed that compared with control groups,UVB was able to induce HIF1α and TfR protein expression in a dose-and time-dependent manner in HaCat cells(P<0.05).TfR mRNA was expressed in a dose-dependent manner and reached a peak at the 8th hour in HaCat cells(P<0.05)whereas HIF-1α mRNA expression was not affected by UVB treatment(P>0.05).The EGFR/PI3K/AKT signaling pathway was required for the induction of HIF-1α and TfR expression induced by UVB.UVB induced activation of EGFR in HaCat cells and EGFR regulated expression of TfR and HIF-1α.EGFR(−/−)MEF did not increase the HIF1 expression following UVB irradiation(P>0.05).In contrast,EGFR(+/+)MEF strongly enhanced HIF1a expression after UVB irradiation(P<0.05).PD153035,a selective inhibitor of EGFR tyrosine kinase,inhibited the TfR protein expression in UVB-treated cells in a dose-dependent manner(P<0.05).PI3K inhibitors,LY294002 and wortmannin,inhibited HIF-1a and TfR expressions induced by UVB(P<0.05).The DEC1(−/−)Ha-Cat cells did not increase their TfR and HIF-1α expressions following UVB irradiation(P>0.05).In contrast,DEC1(+/+)HaCat cells strongly enhanced TfR and HIF-1α protein expression after UVB irradiation(P<0.05).We conclude that UVB induces TfR and HIF-1α expressions via EGFR/PI3K/AKT/DEC1 signaling pathway.

缺铁性贫血孕妇血清可溶性转铁蛋白受体、膜铁转运蛋白1、对氧磷酶1表达水平及检测意义

目的:探讨可溶性转铁蛋白受体(sTfR)、膜铁转运蛋白1(FPN1)、对氧磷酶1(PON1)在缺铁性贫血(IDA)孕妇中的表达及意义。方法:选取IDA孕妇182例为IDA组,根据IDA严重程度将IDA组分为轻度组(80例)、中度组(61例)和重度组(41例),另选取同期健康孕妇73例为健康对照组。比较各组孕妇血清sTfR、FPN1、PON1水平。比较IDA组和健康对照组贫血相关指标。分析血清sTfR、FPN1、PON1水平与血红蛋白(Hb)、平均红细胞体积(MCV)、血清铁蛋白(SF)水平的相关性。绘制受试者工作特征(ROC)曲线分析血清sTfR、FPN1、PON1对孕妇IDA的诊断价值。记录所有孕妇不良结局发生情况,根据最终母婴结局将所有研究对象分为母婴结局正常组(179例)和母婴结局不良组(76例)。比较不同母婴结局孕妇血清sTfR、FPN1、PON1水平。结果:与健康对照组比较,IDA组sTfR、FPN1水平升高,PON1水平降低(均P<0.05)。IDA组Hb、MCV、SF水平低于健康对照组(均P<0.05)。血清sTfR、FPN1与Hb、MCV、SF呈负相关,PON1与Hb、MCV、SF呈正相关(均P<0.05)。血清sTfR、FPN1、PON1对孕妇IDA均有一定诊断效能,且联合检测诊断价值更高(均P<0.05)。重度组血清sTfR、FPN1水平高于轻度和中度组,PON1水平低于轻度和中度组(均P<0.05)。与健康对照组比较,IDA组母婴不良结局总发生率更高(P<0.05)。与母婴结局正常组比较,母婴结局不良组血清sTfR、FPN1水平升高,PON1水平降低(均P<0.05)。结论:血清sTfR、FPN1、PON1在IDA孕妇中均呈异常表达,三者联合检测对IDA诊断效能较高,且与IDA严重程度和母婴结局有关。

铁死亡在不同细菌所致小鼠血流感染模型中的变化规律及生物学意义

背景:发现血流感染新的疾病诊断标志物、治疗疾病及减轻脏器损伤的分子靶点具有重要意义。铁死亡是新发现的一种细胞死亡形式,脓毒症动物模型中铁死亡的过度激活与炎症反应激活以及肝脏、心脏、肾脏等重要脏器的损伤有关,但铁死亡与血流感染的关系尚不十分清楚。目的:探讨铁死亡在不同细菌所致小鼠血流感染模型中的变化规律及生物学意义。方法:建立革兰阴性菌大肠埃希菌、肺炎克雷伯菌及革兰阳性菌金黄色葡萄球菌、粪肠球菌血流感染的SPF级ICR雄性小鼠模型,每组各42只。建模后0.5,1,3,6,12,24,48 h时检测肝脏、心肌、肾脏中铁死亡标志基因转铁蛋白受体1、谷胱甘肽过氧化物酶4 mRNA表达水平。另选用18只SPF级ICR雄性小鼠,随机分为二甲基亚砜(DMSO)对照组、DMSO+肺炎克雷伯菌组、铁死亡抑制剂Ferrostatin-1+肺炎克雷伯菌组,每组6只;后两组采用尾静脉注射肺炎克雷伯菌悬液的方式建立肺炎克雷伯菌血流感染模型,分别在血流感染建模前1 h给予5 mg/kg的Ferrostatin-1及等剂量DMSO腹腔注射;建模后6 h小鼠检测血清中丙氨酸氨基转移酶、天冬氨酸氨基转移酶、血肌酐、血尿素氮、磷酸肌酸激酶同工酶、乳酸脱氢酶以及各组织中铁死亡标志基因的mRNA表达水平。结果与结论:①血流感染建模后,不同细菌血流感染小鼠肝脏、心肌、肾脏中转铁蛋白受体1 mRNA表达水平先升高后降低,谷胱甘肽过氧化物酶4 mRNA表达水平先降低后升高且均在建模后6 h达到峰值;②革兰阴性菌血流感染小鼠中转铁蛋白受体1和谷胱甘肽过氧化物酶4 mRNA表达的变化较革兰阳性菌血流感染小鼠更为显著,其中以肺炎克雷伯菌血流感染小鼠中转铁蛋白受体1和谷胱甘肽过氧化物酶4 mRNA表达的变化最显著;③肺炎克雷伯菌血流感染建模后6 h时,小鼠的丙氨酸氨基转移酶、天冬氨酸氨基转移酶、血肌酐、血尿素氮、磷酸肌酸激酶同工酶、乳酸脱氢酶水平均明显升高;④建模前给予铁死亡抑制剂Ferrostatin-1干预可显著降低丙氨酸氨基转移酶、天冬氨酸氨基转移酶、血肌酐、血尿素氮、磷酸肌酸激酶同工酶、乳酸脱氢酶表达水平;⑤以上结果提示不同细菌致血流感染小鼠中铁死亡明显激活且革兰阴性菌血流感染小鼠的铁死亡激活更为显著;抑制铁死亡可明显减轻肺炎克雷伯菌血流感染小鼠的肝脏、心肌、肾脏损伤。

首发精神分裂症患者血清sTfR、FGF22水平与临床症状的关系及其诊断价值

目的探讨首发精神分裂症(FES)患者血清可溶性转铁蛋白受体(sTfR)及成纤维细胞生长因子22(FGF22)表达情况,分析二者与FES患者临床症状的关系并进行诊断价值分析。方法选取2021年3月至2023年2月在该院确诊的97例FES患者作为FES组,同期选取来该院体检的96例健康志愿者作为对照组。采用免疫透射比浊法检测sTfR水平,酶联免疫吸附试验(ELISA)检测FGF22水平,Spearman法分析FES患者血清中sTfR及FGF22水平与阳性与阴性病症量表(PANSS)评分及威斯康辛卡片分类测验(WCST)结果的相关性,受试者工作特征(ROC)曲线分析sTfR及FGF22水平对FES的临床诊断价值。结果两组在性别、年龄、体质量指数、受教育年限、饮酒史及吸烟史方面差异均无统计学意义(P>0.05),与对照组比较,FES组血清sTfR及FGF22水平均下降(P<0.05)。sTfR单独诊断FES的曲线下面积(AUC)为0.835,最佳截断值为4.606 mg/L,FGF22单独诊断FES的AUC为0.772,最佳截断值为208.333μg/L,二者联合诊断的AUC(0.921)大于sTfR单独诊断的AUC(Z=2.613,P=0.009)及FGF22单独诊断的AUC(Z=5.140,P<0.001);sTfR高水平组及FGF22高水平组的PANSS阳性症状评分、阴性症状评分、病理症状评分、总评分、WCST持续性错误数、错误应答数分别低于sTfR低水平组及FGF22低水平组(P<0.05),而WCST完成分类数、WCST正确应答数分别高于sTfR低水平组及FGF22低水平组(P<0.05);FES组中sTfR、FGF22水平与PANSS阳性症状评分、阴性症状评分、病理症状评分、总评分、WCST持续性错误数、WCST错误应答数均呈负相关(P<0.05),与WCST完成分类数及WCST正确应答数均呈正相关(P<0.05)。结论FES患者血清sTfR及FGF22水平下降,联合检测sTfR及FGF22水平对于FES的临床诊断具有重要意义。

转铁蛋白受体亲和肽衍生物的制备

以2-氯三苯甲基氯(2-CTC)树脂为固相载体、N-芴甲氧羰基(Fmoc)保护氨基酸为原料、N,N′-二异丙基碳二亚胺(DIC)和1-羟基苯并三唑(HOBt)为缩合剂,采用Fmoc固相合成法制备了一种转铁蛋白受体亲和肽(transferrin receptor binding peptide,TfRBP)衍生物,其纯品的HPLC纯度为97.9%,总收率为40.3%,经ESI-MS鉴定其相对分子质量与理论值一致。该方法操作简便、合成周期短、总收率高,为进一步构建TfR靶向多肽-药物偶联物奠定了基础。

靶头修饰对PEG-PCL胶束在人宫颈癌细胞内转运行为的影响

目的 研究转铁蛋白靶向肽T7(7pep)对聚乙二醇-聚己内酯(PEG-PCL)胶束在人宫颈癌HeLa细胞内转运行为的影响。方法 以香豆素-6(C6)为荧光指示探针,通过薄膜分散法制备包载有C6的PEG-PCL(PEG-PCL-C6)胶束以及靶头7pep修饰的PEG-PCL(7pep-PEG-PCL-C6)胶束。比较两种胶束的粒径、多分散指数及外观形态;比较两种胶束被HeLa细胞实时摄取的情况及其入胞后与早期内吞体(EE)、内吞循环室(ERC)、晚期内吞体(LE)的共定位情况。结果 PEG-PCL-C6和7pep-PEG-PCL-C6胶束的平均粒径分别为(75.0±2.3)、(82.0±1.5)nm,多分散指数分别为0.17±0.20、0.17±0.32,外观均为规整的圆球形。7pep-PEGPCL-C6胶束的入胞速度和入胞量均明显快/多于PEG-PCL-C6胶束。7pep-PEG-PCL-C6胶束比PEG-PCL-C6胶束能够更快地进入EE,而入胞后PEG-PCL-C6胶束进入ERC的速率较7pep-PEG-PCL-C6胶束快,且PEG-PCL-C6胶束和7pep-PEG-PCL-C6胶束在LE均有逐渐累积的趋势,但7pep-PEG-PCL-C6胶束入胞60 min时与LE的皮尔森系数、信号重叠比率、共定位比率均显著低于入胞30 min时(P<0.05或P<0.01)。结论 靶头7pep修饰可提高PEG-PCL-C6胶束的入胞速率和入胞量,还可改变其胞内转运行为。

hnRNPA2B1通过抑制转铁蛋白受体增强胰腺癌细胞对铁死亡的抵抗

目的:探讨异质核核糖核蛋白A2B1(heterogeneous nuclear ribonucleoprotein A2B1,hnRNPA2B1)对胰腺癌细胞铁死亡的影响及其潜在机制。方法:将3种胰腺癌PaTu8988、MIA-paca2、PANC1细胞用不同浓度Erastin处理3 d,CCK8法检测细胞活性,计算3种细胞Erastin半数抑制浓度(IC_(50))并比较其对Erastin的抵抗力;荧光实时定量PCR(qRT-PCR)和蛋白免疫印迹法分别检测3种胰腺癌细胞中hnRNPA2B1 mRNA和蛋白的相对表达。通过GEPIA平台分析hnRNPA2B1在胰腺癌组织和正常组织中的差异性表达,通过GEO数据库下载临床数据集分析差异性表达hnRNPA2B1患者的预后。在PaTu8988细胞中干扰hnRNPA2B1表达,分别转染sh-Control、sh-hnRNPA2B1质粒;在PANC1细胞中过表达hnRNPA2B1,分别转染Vector、Flag-hnRNPA2B1质粒;分别采用qRT-PCR和蛋白免疫印迹检测转染效率,通过Transwell实验检测hnRNPA2B1对胰腺癌细胞迁移和侵袭的影响,通过CCK8法检测hnRNPA2B1对胰腺癌细胞增殖的影响;对sh-Control组和sh-hnRNPA2B1组、Vector组和Flag-hnRNPA2B1组,分别予以对照(0μmol/L Erastin)、Erastin(IC_(50)浓度)处理,通过流式细胞术检测脂质过氧化物含量,比色法检测丙二醛和组织铁含量,此外sh-hnRNPA2B1组和Flag-hnRNPA2B1组加入Erastin的同时加入铁死亡挽救剂Ferrostatin-1,坏死挽救剂Necrostatin-1或凋亡挽救剂ZVAD-FMK,台酚蓝染色法检测细胞活性;qRT-PCR和蛋白免疫印迹法分别检测胰腺癌细胞转铁蛋白受体(transferrin receptor,TFRC)、铁蛋白重链、谷胱甘肽过氧化物酶4、溶质载体家族7成员11等mRNA和蛋白相对表达水平。结果:3种胰腺癌细胞对Erastin抗性由高到低依次是PaTu8988、MIA-paca2、PANC1细胞,其Erastin IC_(50)依次为18.020、15.760、2.947μmol/L;hnRNPA2B1表达量由高到低的趋势与Erastin IC_(50)趋势相同,于PaTu8988细胞中最高,MIA-paca2细胞中次之,PANC1细胞中最低(P均<0.05);胰腺癌组织中hnRNPA2B1表达水平明显高于正常组织,高表达hnRNPA2B1患者预后较差(P均<0.05)。下调hnRNPA2B1表达后,PaTu8988细胞迁移、侵袭和增殖能力明显降低,上调则与之相反(P均<0.05)。经Erastin处理后sh-hnRNPA2B1组和Flag-hnRNPA2B1组降低的细胞活性仅可被Ferrostatin-1挽救(P<0.05);与sh-Control组相比,sh-hnRNPA2B1组脂质过氧化物、丙二醛和组织铁含量明显增高(P均<0.05);与Vector组相比,Flag-hnRNPA2B1组脂质过氧化物、丙二醛和组织铁含量明显降低(P均<0.05)。hnRNPA2B1可抑制TFRC表达,干扰hnRNPA2B1则与之相反(P均<0.05)。结论:hnRNPA2B1通过抑制TFRC表达阻止铁蓄积,从而促进胰腺癌细胞铁死亡抵抗。

双罐串联连续血液净化对慢性肾衰竭尿毒症患者血清可溶性转铁蛋白受体和人巨噬细胞趋化蛋白-1水平及生存质量的影响

目的:探讨双罐串联连续血液净化(CRRT)对慢性肾衰竭尿毒症患者血清可溶性转铁蛋白受体(sTfR)和人巨噬细胞趋化蛋白-1(MCP-1)水平及生存质量的影响。方法:选择2022年4月至2023年4月新疆医科大学第一附属医院行血液透析治疗的96例慢性肾衰竭尿毒症患者,随机分为单罐组(CRRT连续血液净化单个灌流器)和双罐串联组(CRRT连续血液净化2个灌流器串联),每组48例。分别于治疗前,治疗后2个月、半年及1年时检测两组患者血清sTfR和MCP-1水平,判断其治疗后皮肤瘙痒缓解程度;分别于治疗前及治疗后72h记录两组白细胞水平、急性生理与慢性健康量表(APACHEⅡ)评分情况。采用中文版欧洲五维量表(CEQ-5D-3L)、视觉模拟量表(VAS)评分,评估两组患者治疗前后生命质量。结果:两组治疗前血清sTfR水平比较差异无统计学意义(P>0.05)。治疗后2个月、半年及1年各时间点,双罐串联组血清sTfR水平均低于单罐组,差异有统计学意义(t=5.089、18.410、41.306,P<0.05);两组治疗前血清MCP-1水平比较,差异无统计学意义(P>0.05)。治疗后2个月、半年及1年各时间点,双罐串联组血清MCP-1水平均低于单罐组,差异有统计学意义(t=8.554、8.019、10.744,P<0.05);两组治疗后原有皮肤瘙痒程度均有所改善,但单罐组缓解程度明显低于双罐串联组,差异有统计学意义(x^(2)=8.540,P<0.05);两组治疗前白细胞、APACHEⅡ评分比较,差异无统计学意义(P>0.05),治疗后72h白细胞、APACHEⅡ评分均降低,且双罐串联组改善程度均优于单罐组,差异有统计学意义(t=5.549、14.781,P<0.05);两组治疗前血清CEQ-5D-3L评分比较,差异无统计学意义(P>0.05)。治疗后2个月、半年及1年各时间点,双罐串联组健康描述系统标准总分均低于单罐组,VAS评分高于单罐组,差异有统计学意义(t健康描述系统标准总分=4.744、5.103、9.418,tVAS评分=3.375、2.866、3.126,P<0.05)。结论:双罐串联连续血液净化可有效降低慢性肾衰竭尿毒症患者血清sTfR,MCP-1水平,缓解皮肤瘙痒程度,提高生存质量,同时还能有效降低白细胞水平,改善APACHEⅡ评分和血常规。