ITGB2 fosters the cancerous characteristics of ovarian cancer cells through its role in mitochondrial glycolysis transformation.

Related studies have shown that ITGB2 mediates mitochondrial glycolytic transformation in cancer-associated fibroblasts and participates in tumor occurrence, metastasis and invasion of cancer cells. Based on these studies, we tried to construct a mitochondrial glycolysis regulatory network and explored its effect on mitochondrial homeostasis and ovarian cancer cells' cancerous characteristics. Our research revealed a distinct increase in the expression of ITGB2 and associated signaling pathway elements (PI3K-AKT-mTOR) in cases of ovarian cancer. ITGB2 might control mTOR expression via the PI3K-AKT pathway, thus promote mitochondrial glycolysis transformation and cell energy supply in ovarian cancer. This pathway could also inhibit mitophagy, maintain mitochondrial stability, and enhance the cancerous characteristics in case of ovarian cancer cells by mediating mitochondrial glycolytic transformation. Thus, we concluded that ITGB2-associated signaling route (PI3K-AKT-mTOR) may contribute to the progression of cancerous traits in ovarian cancer via mediating mitochondrial glycolytic transformation.

A meta-analysis of the prognostic impact of tissue golgi protein 73 (tGP73) in hepatocellular carcinoma.

INTRODUCTION: To date, an increasing number of studies have revealed that GP73 may have prognostic value in liver cancer. However, most of the studies evaluated serum GP73, and the results regarding the prognostic value of tGP73 in liver cancer are still controversial. Therefore, in this meta-analysis, we aimed to determine whether tGP73 has any prognostic value in patients with HCC. MATERIALS AND METHODS: Relevant publications were searched for in PubMed, EMBASE, OVID, the Cochrane Library, and the Web of Science databases up to March 2023. The hazard ratio (HR) or odds ratio (OR) with corresponding 95% confidence intervals (95% CIs) of eligible studies were assessed by fixed-effects or random-effects models. In addition, subgroup analyses were conducted to investigate the possible causes of heterogeneity, and publication bias analysis was also performed to assess the reliability of the meta-analysis results. RESULTS: A total of 10 studies were included. These studies included 1569 HCC patients, and a meta-analysis was performed. The results of our meta-analysis showed that higher GP73 expression levels were significantly associated with poorer OS (HR = 1.87, 95% CI: 1.41-2.48, P < 0.0001, I2 = 58%). However, there was no significant correlation between high GP73 expression and disease-free survival (DFS) (HR: 1.43, 95% CI: 0.93-2.33, P = 0.100). In addition, abnormal GP73 expression was also related to higher tumour tissue differentiation grade (OR = 3.03, 95% CI = 2.01-4.57, P < 0.0001, I2 = 89%), later tumour stage (OR = 5.89, 95% CI = 2.31-14.99, P < 0.0001, I2 = 0%), vascular invasion (OR = 1.72, 95% CI = 1.12-2.64, P = 0.010, I2 = 0%), multiple tumours (OR = 2.44, 95% CI = 1.37-3.68, P = 0.001, I2 = 44%) and early postoperative tumour recurrence (OR = 1.92, 95% CI = 1.10-3.28, P = 0.020, I2 = 62%). CONCLUSIONS: The meta-analysis showed that the overexpression of GP73 may be related to a poor prognosis of HCC, and it may also have a predictive effect on the invasion and metastasis of HCC.

Overexpression miR-520a-3p inhibits acute myeloid leukemia progression via targeting MUC1.

BACKGROUND: Acute myeloid leukemia (AML) is one of the familiar malignant tumors in the hematological system. miR-520a-3p is reported to be involved in several cancers' progression. However, miR-520a-3p role in AML remains unclear. In this study, we aimed to clarify the role and potential mechanism of miR-520a-3p in AML. METHODS: Cell viability, proliferation, cycle and apoptosis were detected by MTT assay, colony formation assay, flow cytometry, respectively. The levels of PNCA, Bcl-2, Cleaved caspase 3, Cleaved caspase 9 and ß-catenin protein were detected by Western blot. Dual-luciferase reported assay was performed to detect the regulation between miR-520a-3p and MUC1. To verify the effect of miR-520a-3p on tumor proliferation in vivo, a non-homogenous transplant model of tumors was established. RESULTS: miR-520a-3p expression was down-regulated, and MUC1 expression was up-regulated in AML patients. miR-520a-3p overexpression suppressed THP-1 cell proliferation, induced cell cycle G0/G1 inhibition and promoted apoptosis. miR-520a-3p targeted MUC1 and negatively regulated its expression. MUC1 knockdown inhibited THP-1 cell proliferation and promoted apoptosis. miR-520a-3p overexpression inhibited AML tumors growth. CONCLUSION: Overexpression miR-520a-3p inhibited AML cell proliferation, and promoted apoptosis via inhibiting MUC1 expression and repressing Wnt/ß-catenin pathway activation.

A CpG Methylation Classifier to Predict Relapse in Adults with T-Cell Lymphoblastic Lymphoma.

PURPOSE: Adults with T-cell lymphoblastic lymphoma (T-LBL) generally benefit from treatment with acute lymphoblastic leukemia (ALL)-like regimens, but approximately 40% will relapse after such treatment. We evaluated the value of CpG methylation in predicting relapse for adults with T-LBL treated with ALL-like regimens. EXPERIMENTAL DESIGN: A total of 549 adults with T-LBL from 27 medical centers were included in the analysis. Using the Illumina Methylation 850K Beadchip, 44 relapse-related CpGs were identified from 49 T-LBL samples by two algorithms: least absolute shrinkage and selector operation (LASSO) and support vector machine-recursive feature elimination (SVM-RFE). We built a four-CpG classifier using LASSO Cox regression based on association between the methylation level of CpGs and relapse-free survival in the training cohort (n = 160). The four-CpG classifier was validated in the internal testing cohort (n = 68) and independent validation cohort (n = 321). RESULTS: The four-CpG-based classifier discriminated patients with T-LBL at high risk of relapse in the training cohort from those at low risk (P < 0.001). This classifier also showed good predictive value in the internal testing cohort (P < 0.001) and the independent validation cohort (P < 0.001). A nomogram incorporating five independent prognostic factors including the CpG-based classifier, lactate dehydrogenase levels, Eastern Cooperative Oncology Group performance status, central nervous system involvement, and NOTCH1/FBXW7 status showed a significantly higher predictive accuracy than each single variable. Stratification into different subgroups by the nomogram helped identify the subset of patients who most benefited from more intensive chemotherapy and/or sequential hematopoietic stem cell transplantation. CONCLUSIONS: Our four-CpG-based classifier could predict disease relapse in patients with T-LBL, and could be used to guide treatment decision.

A gene-expression-based signature predicts survival in adults with T-cell lymphoblastic lymphoma: a multicenter study.

We aimed to establish a discriminative gene-expression-based classifier to predict survival outcomes of T-cell lymphoblastic lymphoma (T-LBL) patients. After exploring global gene-expression profiles of progressive (n = 22) vs. progression-free (n = 28) T-LBL patients, 43 differentially expressed mRNAs were identified. Then an eleven-gene-based classifier was established using LASSO Cox regression based on NanoString quantification. In the training cohort (n = 169), high-risk patients stratified using the classifier had significantly lower progression-free survival (PFS: hazards ratio 4.123, 95% CI 2.565-6.628; p < 0.001), disease-free survival (DFS: HR 3.148, 95% CI 1.857-5.339; p < 0.001), and overall survival (OS: HR 3.790, 95% CI 2.237-6.423; p < 0.001) compared with low-risk patients. The prognostic accuracy of the classifier was validated in the internal testing (n = 84) and independent validation cohorts (n = 360). A prognostic nomogram consisting of five independent variables including the classifier, lactate dehydrogenase levels, ECOG-PS, central nervous system involvement, and NOTCH1/FBXW7 status showed significantly greater prognostic accuracy than each single variable alone. The addition of a five-miRNA-based signature further enhanced the accuracy of this nomogram. Furthermore, patients with a nomogram score ≥154.2 significantly benefited from the BFM protocol. In conclusion, our nomogram comprising the 11-gene-based classifier may make contributions to individual prognosis prediction and treatment decision-making.

[Transcription factors analysis of subchondral bone in early experimental osteoarthritis based on gene expression profiles].

OBJECTIVE: To identify the master transcription factors (TF) that might be responsible for the gene expression alteration of OA. METHODS: Raw expression data for rat OA model(GSE30322) was downloaded from NCBI GEO database. Microarray data analysis for rat and human was carried out separately using functions from limma packagein R, gene expression was considered as significantly changed between conditions if adjusted P-value<0.05 and the absolute value of fold change>=2. iRegulon was applied to differentially up-regulated and down-regulated genes in OA separately. RESULTS: (1)15 TFs, including FOXN4, NANOS1, E2F6, RAD21, MECOM, ETS1, MEF2A, POU2F3, BRCA1, GATA3, ZNF706, ZBTB33, SUZ12, DBP and SETDB1, were identified as the potential master TFs of up-regulated DEGs with statistical significance. (2)12 TFs, including ARID3A, YY1, RDBP, ATF1, CRX, TAF1, XBP1, SOX3, E2F4, PGR, TIMM8A and HOXA2, were identified as the potential master TFs of down-regulated DEGs with statistical significance. CONCLUSIONS: The newly identified TFs maybe play important roles in pathogenesis of early experimental osteoarthritis, and our study provides new diagnostic markers or therapeutic targets for OA.

Percutaneous transhepatic biliary drainage and stenting for malignant obstructive jaundice: A report of two cases.

Malignant obstructive jaundice comprises a group of diseases that can be caused by primary biliary and extra-biliary carcinomas. Generally, surgical resection is the primary treatment for malignant obstructive jaundice; however, for the patients that are unable to undergo surgery, urgent treatment is required to improve hepatic function. Percutaneous transhepatic biliary drainage (PTBD) and stenting are emerging alternative treatments for malignant obstructive jaundice. PTBD and stenting have exhibited good efficacy for the treatment of malignant obstructive jaundice, with few complications and reduced associated pain.

Clinical Characteristics and Treatment of Trigeminal Neuralgia Following Herpes Zoster.

OBJECTIVE: The aim of this study was to illustrate the clinical characteristics and treatment of trigeminal neuralgia following herpes zoster. METHODS: From August 1, 2011 to August 1, 2013, 23 consecutive patients with trigeminal neuralgia following herpes zoster underwent microvascular decompression (MVD) at our cranial nerve disease center. All patients underwent preoperative MRI evaluation, intraoperative observation, and clinical effect evaluation. Clinical data were collected and analyzed in our center. RESULTS: V2 division was the most commonly affected branch. Unlike pretrigeminal neuralgia (PTN), trigger zone was only found in a small part of patients (21.7%). Unlike PTN, the adhesions and compressions between trigeminal nerve and offending vessels were usually not serious; trigeminal nerve usually is atrophic; superior cerebellar artery was the most common offending vessels (65.2%). Of 23 patients, 19 experienced pain relief (82.6%), 1 patient suffered from hearing loss, and another one suffered from cerebrospinal fluid leak; no severe complications were found. During follow-up period, no recurrence was found (3 lost). CONCLUSIONS: For patients who suffered from trigeminal neuralgia following herpes zoster, trigger zone was only found in a small part of patients. The trigeminal nerve usually is atrophic; microvascular decompression was equally applied to these patients if vessel compression was confirmed.

Epidemiological characteristics of human rabies in Henan province in China from 2005 to 2013

Background Rabies is very common in Peoples Republic of China. Henan province, in the central portion of China, is one of the most densely populated provinces in the entire country. With the new rabies epidemic trend noted in northern and western China, it is necessary to investigate the characteristics of human rabies in this area and control the disease.Methods We chose patients in hospital isolation in 18 municipalities in Henan province as the investigation subjects. Data were collected through systematic reporting from these hospitals, whereas a questionnaire was applied to the relatives of patients.Results A total of 1022 rabies cases were reported. The incidence of human rabies in Henan has increased rapidly since 2005, having peaked in 2007, and maintained a high level in the remaining years. The cases were found mainly in rural areas in the south and east of the province. Rabies was often noted in summer and with the highest number in August. Most cases were noted in males and often in farmers. The patients aged between 40 and 60 years accounted for 36.8 % of all the cases. The wound treatment rate (12.2 %) and vaccination rate (2.6 %) of rabies cases after exposure were relatively low, while the rabies immunoglobulin utilization rate was only 2.8 %.Conclusions Rabies epidemic cases at the county level overall were increasing in Henan province during the period of 20052013; the epidemic has spread quickly. The data in this study imply that the disease could be better managed by more integrated surveillance across human health and veterinary sectors, improved education and better government policies.

Epidemiological characteristics of human rabies in Henan province in China from 2005 to 2013

Background Rabies is very common in People's Republic of China. Henan province, in the central portion of China, is one of the most densely populated provinces in the entire country. With the new rabies epidemic trend noted in northern and western China, it is necessary to investigate the characteristics of human rabies in this area and control the disease.Methods We chose patients in hospital isolation in 18 municipalities in Henan province as the investigation subjects. Data were collected through systematic reporting from these hospitals, whereas a questionnaire was applied to the relatives of patients.Results A total of 1022 rabies cases were reported. The incidence of human rabies in Henan has increased rapidly since 2005, having peaked in 2007, and maintained a high level in the remaining years. The cases were found mainly in rural areas in the south and east of the province. Rabies was often noted in summer and with the highest number in August. Most cases were noted in males and often in farmers. The patients aged between 40 and 60 years accounted for 36.8 % of all the cases. The wound treatment rate (12.2 %) and vaccination rate (2.6 %) of rabies cases after exposure were relatively low, while the rabies immunoglobulin utilization rate was only 2.8 %.Conclusions Rabies epidemic cases at the county level overall were increasing in Henan province during the period of 2005-2013; the epidemic has spread quickly. The data in this study imply that the disease could be better managed by more integrated surveillance across human health and veterinary sectors, improved education and better government policies.(AU)

Surgical skills of adhesions and transposition of trigeminal nerve for primary trigeminal neuralgia.

OBJECTIVE: The aims of this study were to investigate disposal of adhesions and transposition of the trigeminal nerve during microvascular decompression and evaluate its surgical effect. METHODS: By collecting the clinical data of typical trigeminal neuralgia patients we treated from January 2013 to May 2013, we chose 120 patients with adhesions and transposition of trigeminal nerve, analyzed their preoperative imaging features and surgical procedures, and evaluated postoperative effect after 3-month follow-up. RESULTS: Among these 120 patients, 113 cases showed positive effect in magnetic resonance imaging (three-dimensional time-of-flight); the positive rate was 94.2%. During the operation, firstly we proceed to separate the nerve and artery, secondly we released the vascular compression, finally we removed the superior cerebellar artery to the nerve's head end. Postoperative effect evaluation stated that 101 patients showed immediate remission and 13 patients showed delayed remission (3 min after operation) and the surgery was ineffective for 6 patients; the remission rate was 95%. There were no severe complications. CONCLUSIONS: Magnetic resonance imaging (three-dimensional time-of-flight) before operation could clearly display the neurovascular relationship of trigeminal neuralgia patients. It offers great help for preoperative evaluation. Separation between arachnoid, nerve, and artery; vascular decompression; and removal of the superior cerebellar artery to the head end could resolve adhesions and transposition of trigeminal nerve during microvascular decompression, which showed significant effects.

Clinical characteristics and surgical techniques of trigeminal neuralgia caused simply by venous compression.

OBJECTIVE: The objective of this study was to introduce the operation essentials for treatment of patients with vein compression so as to obtain a satisfactory decompression without sacrificing veins. METHODS: We chose 15 patients with trigeminal neuralgia caused by venous from June 15, 2010, through June 15, 2011, and performed microvascular decompression for each patient. By collecting clinical data, such as preoperative magnetic resonance imaging scans, key operative procedures, surgical outcomes, and complications, we explored the operation techniques for these patients and finally summarized our experiences and ideas. RESULTS: For all the 15 patients, 9 cases had excellent remission, 3 cases had delayed excellent remission, 1 case had good remission, and 2 cases had failed result; the total remission rate was 86.7%; 3 cases had facial numbness. CONCLUSIONS: For patients with vein compression, we combine the following 4 procedures together: (1) fully releasing the arachnoid around trigeminal nerve, (2) exploration and decompression of the whole trigeminal root from Meckel cave to pons, (3) cauterization of companioned petrosal vein tributaries by bipolar coagulation, and (4) placing Teflon between trigeminal nerve and offending petrosal vein, thus could acquire a satisfactory effect (remission rate, 86.7%).

Albimonas pacifica sp. nov., isolated from seawater of the Pacific, and emended description of the genus Albimonas.

A Gram-stain-negative, aerobic, catalase- and oxidase-positive, non-flagellated, rod-shaped bacterium, designated strain P-50-3(T), was isolated from seawater of the Pacific. The strain grew at 10-40 °C (optimum at 30 °C) and with 0-12 % (w/v, optimum 2 %) NaCl. It reduced nitrate to nitrite but did not hydrolyse gelatin, starch or Tween 80. Analysis of 16S rRNA gene sequences showed that strain P-50-3(T) clustered tightly with the genus Albimonas and shared the highest 16S rRNA gene sequence similarity (94.3 %) with the type strain of Albimonas donghaensis. The major respiratory quinone was Q-10 and the major cellular fatty acids were C18 : 1ω7c, C18 : 0, 11-methyl C18 : 1ω7c and C16 : 0. Polar lipids included phosphatidylglycerol (PG), phosphatidylcholine (PC), two unidentified aminolipids and an unidentified lipid. The genomic DNA G+C content of strain P-50-3(T) was 69.0 mol%. On the basis of the data obtained in this polyphasic study, strain P-50-3(T) represents a novel species within the genus Albimonas, for which the name Albimonas pacifica sp. nov. is proposed. The type strain of Albimonas pacifica is P-50-3(T) ( = KACC 16527(T) = CGMCC 1.11030(T)). An emended description of the genus Albimonas Lim et al. 2008 is also proposed.

Arenitalea lutea gen. nov., sp. nov., a marine bacterium of the family Flavobacteriaceae isolated from intertidal sand.

A yellow, rod-shaped, Gram-negative, facultatively aerobic, gliding bacterium, designed strain P7-3-5(T), was isolated from intertidal sand of the Yellow Sea, China. Analysis of 16S rRNA gene sequences revealed that strain P7-3-5(T) formed a distinct lineage within the family Flavobacteriaceae, sharing 94.2-96.9 % sequence similarity with type strains of species of the most closely related genera, including Hyunsoonleella, Jejuia, Marinivirga and Algibacter. The strain grew at 4-40 °C and with 0.5-5.0 % (w/v) NaCl. It reduced nitrate to nitrite and hydrolysed gelatin and DNA. The major cellular fatty acids were iso-C15 : 0, iso-C15 : 1 G and anteiso-C15 : 0 and the major respiratory quinone was MK-6. Polar lipids included phosphatidylethanolamine (PE), three unidentified aminolipids (AL1-3) and four unidentified lipids (L1-4). The genomic DNA G+C content of strain P7-3-5(T) was 32.1 mol%. Data from this polyphasic study suggest that strain P7-3-5(T) represents a novel species in a new genus in the family Flavobacteriaceae, for which the name Arenitalea lutea gen. nov., sp. nov. is proposed. The type strain of Arenitalea lutea is P7-3-5(T) ( = CGMCC 1.12213(T) = KACC 16457(T)).

Pseudorhodobacter antarcticus sp. nov., isolated from Antarctic intertidal sandy sediment, and emended description of the genus Pseudorhodobacter Uchino et al. 2002 emend. Jung et al. 2012.

A Gram-negative, aerobic, non-motile, pink-pigmented and rod-shaped strain, designated ZS3-33(T), was isolated from Antarctic intertidal sandy sediment. The strain grew optimally at 15 °C and with 1.0 % (w/v) NaCl. It reduced nitrate to nitrite and hydrolysed Tween 20. It could not produce bacteriochlorophyll a. The predominant cellular fatty acid was C18 : 1ω7c and the predominant respiratory quinone was Q-10. The major polar lipids were phosphatidylglycerol, phosphatidylcholine, two unidentified aminophospholipids and an unidentified aminolipid. Analyses of 16S rRNA gene sequences revealed that strain ZS3-33(T) belonged to the genus Pseudorhodobacter, showing 97.4 % similarity to the type strain of Pseudorhodobacter ferrugineus and 95.3 % similarity to the type strain of Pseudorhodobacter aquimaris. Levels of gyrB gene sequence similarity between strain ZS3-33(T) and the type strains of P. ferrugineus and P. aquimaris were 87.6 and 81.7 %, respectively. DNA-DNA relatedness between strain ZS3-33(T) and P. ferrugineus DSM 5888(T) was 56.6 %. The genomic DNA G+C content of strain ZS3-33(T) was 57.1 mol%. Based on data from this polyphasic study, strain ZS3-33(T) represents a novel species of the genus Pseudorhodobacter, for which the name Pseudorhodobacter antarcticus sp. nov. is proposed. The type strain is ZS3-33(T) ( = CGMCC 1.10836(T) = KCTC 23700(T)). An emended description of the genus Pseudorhodobacter Uchino et al. 2002 emend. Jung et al. 2012 is also proposed.

Marinicauda pacifica gen. nov., sp. nov., a prosthecate alphaproteobacterium of the family Hyphomonadaceae isolated from deep seawater.

A marine prosthecate bacterium, designated strain P-1 km-3(T), was isolated from deep seawater from the Pacific. Cells of strain P-1 km-3(T) were Gram-stain-negative, aerobic, catalase- and oxidase-positive, dimorphic rods with a single polar prostheca or flagellum. The strain hydrolysed gelatin and grew at 6-40 °C (optimum, 30 °C) and with 0.5-12% (w/v) NaCl (optimum, 2%). Phylogenetic analysis of the 16S rRNA gene sequences revealed that strain P-1 km-3(T) belonged to the family Hyphomonadaceae in the class Alphaproteobacteria and represented a separate lineage, located between the genera Oceanicaulis and Woodsholea. Sequence similarities of strain P-1 km-3(T) with type strains of species of the genera Oceanicaulis and Woodsholea were 93.2-93.9%. The predominant cellular fatty acids in strain P-1 km-3(T) were C18:1ω7c, C18:0, 11-methyl C18:1ω7c, C17:0 and C19:0 cyclo ω8c. The major respiratory quinone of strain P-1 km-3(T) was Q-10. The polar lipids of strain P-1 km-3(T) comprised glucuronopyranosyldiglyceride (GUDG), monoglycosyldiglyceride (MGDG), sulfo-quinovosyl diacylglycerol (SQDG), phosphatidylglycerol (PG), an unidentified phospholipid (PL) and an unidentified lipid (L). The genomic DNA G+C content of strain P-1 km-3(T) was 66.0 mol%. On the basis of the polyphasic data presented in this study, strain P-1 km-3(T) is proposed to represent a novel species in a new genus, Marinicauda pacifica gen. nov., sp. nov., within the family Hyphomonadaceae. The type strain of the type species is P-1 km-3(T) (=KACC 16526(T)=CGMCC 1.11031(T)).

Marinobacter antarcticus sp. nov., a halotolerant bacterium isolated from Antarctic intertidal sandy sediment.

A Gram-staining-negative, aerobic, motile, oxidase- and catalase-positive, rod-shaped strain, designated ZS2-30(T), was isolated from Antarctic intertidal sandy sediment. The strain grew at 4-35 °C (optimum, 25 °C) and in 0-25% (w/v) NaCl (optimum, 3.0-4.0%). It could reduce nitrate to nitrite and hydrolyse Tween 80. The predominant cellular fatty acids of strain ZS2-30(T) were summed feature 3 (C(16:1)ω7c and/or C(16:1)ω6c), C(16:0), C(18:1)ω9c, C(16:1)ω9c, C(12:0) 3-OH and C(12:0). The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unidentified aminophospholipid. The genomic DNA G+C content of strain ZS2-30(T) was 55.8 mol%. Analyses of 16S rRNA gene sequences revealed that strain ZS2-30(T) was affiliated with the genus Marinobacter. It showed highest 16S rRNA gene sequence similarities to the type strains of three species of the genus Marinobacter, namely Marinobacter maritimus (98.3%), Marinobacter psychrophilus (98.1%) and Marinobacter goseongensis (97.1%), but the DNA-DNA relatedness values between strain ZS2-30(T) and the above three species were all lower than 45%. Moreover, strain ZS2-30(T) could be distinguished from closely related species of the genus Marinobacter by various phenotypic properties. Based on this taxonomic study using a polyphasic approach, strain ZS2-30(T) is considered to represent a novel species in the genus Marinobacter, for which the name Marinobacter antarcticus sp. nov. is proposed. The type strain of Marinobacter antarcticus is ZS2-30(T) ( = CGMCC 1.10835(T) = KCTC 23684(T)).

Immunization with truncated sequence of Telomerase Reverse Transcriptase induces a specific antitumor response in vivo.

To select the MHC-I-binding epitope-rich sequence of mice telomerase reverse transcriptase (mTERT) and study the antitumor immune response induced by truncated TERT through mRNA-transfected dendritic cells (DCs) immunization in mice. The MHC-I-binding epitopes of TERT were predicted using bioinformatics software. The selected sequence of TERT (Truncated mTERT, TERT(t), mTERT cDNA 1776 bp-2942 bp encoding 584 aa-969 aa) was cloned from B16 mouse melanoma cells and inserted into pBluescriptIIKS(+) plasmid downstream of the T7 promoter. TERT(t) RNA was prepared through in vitro transcription. Bone marrow-derived DCs were electroporated with TERT(t) RNA and used to immunize syngeneic naïve mice. The quantity and cytotoxic activity of TERT-specific cytotoxic T lymphocytes (CTLs) in mice spleen were evaluated using IFN-gamma enzyme-linked immunospot (ELISPOT) and Lactate dehydrogenase release assay. The immunoprophylactic effects against TERT positive tumor induced by TERT(t) RNA transfected DC in vivo were evaluated through an immunized-challenged mouse model. TERT(t) was cloned and in vitro transcribed into TERT(t) mRNA. As shown in FCM analysis, the efficiency of DC electroporation is 35.1% (29.7-41.2%). After electroporation, a subtle increase of costimulator and MHC-II molecules were expressed on the cell surface. Immunization of TERT(t) mRNA transfected DCs induced IFN-gamma-secreting CTLs which manifested specific cytotoxic activity against TERT-positive target cells. In a cancer mouse model, vaccination of TERT(t) mRNA-transfected DCs suppressed the growth of TERT positive tumors (p=0.001) and prolong the survival time of tumor-bearing animals (p=0.029). TERT(t) evokes an antitumor immune response in vivo which is targeted to TERT. TERT(t) can be used as an antigeneic sequence to produce anti-TERT tumor vaccine.

Heat-shocked tumor cell lysate-pulsed dendritic cells induce effective anti-tumor immune response in vivo.

AIM: To study whether heat-shocked tumor cells could enhance the effect of tumor cell lysate-pulsed dendritic cells (DCs) in evoking anti-tumor immune response in vivo. METHODS: Mouse undifferentiated colon cancer cells (CT-26) were heated at 42 degrees Celsius for 1 h and then frozen-thawed. The bone marrow-derived DCs pulsed with heat-shocked CT-26 cell lysate (HSCT-26 DCs) were recruited to immunize syngeneic naive BALB/c mice. The cytotoxic activity of tumor specific cytotoxic T lymphocytes (CTLs) in mouse spleen was evaluated by IFN-enzyme-linked immunospot (ELISpot) and LDH release assay. The immunoprophylactic effects induced by HSCT-26 DCs in mouse colon cancer model were compared to those induced by single CT-26 cell lysate-pulsed DCs (CT-26 DCs) on tumor volume, peritoneal metastasis and survival time of the mice. RESULTS: Heat-treated CT-26 cells showed a higher hsp70 protein expression. Heat-shocked CT-26 cell lysate pulsing elevated the co-stimulatory and MHC-II molecule expression of bone marrow-derived DCs as well as interleukin-12 p70 secretion. The IFN-gamma secreting CTLs induced by HSCT-26 DCs were significantly more than those induced by CT-26 DCs (P=0.002). The former CTLs' specific cytotoxic activity was higher than the latter CTLs' at a serial E/T ratio of 10:1, 20:1, and 40:1. Mouse colon cancer model showed that the tumor volume of HSCT-26 DC vaccination group was smaller than that of CT-26 DC vaccination group on tumor volume though there was no statistical difference between them (24 mm3 vs 8 mm3, P=0.480). The median survival time of mice immunized with HSCT-26 DCs was longer than that of those immunized with CT-26 DCs (57 d vs 43 d, P=0.0384). CONCLUSION: Heat-shocked tumor cell lysate-pulsed DCs can evoke anti-tumor immune response in vivo effectively and serve as a novel DC-based tumor vaccine.