Participation of CCL1 in Snail-Positive Fibroblasts in Colorectal Cancer Contribute to 5-Fluorouracil/Paclitaxel Chemoresistance / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Cancer-associated fibroblasts (CAFs) activated by cancer cells has a central role in development and malignant biological behavior in colorectal cancer (CRC). Adult fibroblasts do not express Snail, but Snail-positive fibroblasts are discovered in the stroma of malignant CRC and reported to be the key role to chemoresistance. However, the reciprocal effect of CAFs expressed Snail to chemoresistance on CRC cells and the underlying molecular mechanisms are not fully characterized. MATERIALS AND METHODS: Snail-overexpressed 3T3 stable cell lines were generated by lipidosome and CT26 mixed with 3T3-Snail subcutaneous transplanted CRC models were established by subcutaneous injection. Cell Counting Kit-8, flow cytometry and western blotting assays were performed, and immunohistochemistry staining was studied. The cytokines participated in chemoresistance was validated with reverse transcriptase-polymerase chain reaction and heatmap. RESULTS: Snail-expression fibroblasts are discovered in human and mouse spontaneous CRCs. Overexpression of Snail induces 3T3 fibroblasts transdifferentiation to CAFs. CT26 co-cultured with 3T3-Snail resisted the impairment from 5-fluorouracil and paclitaxel in vitro. The subcutaneous transplanted tumor models included 3T3-Snail cells develop without restrictions even after treating with 5-fluorouracil or paclitaxel. Moreover, these chemoresistant processes may be mediated by CCL1 secreted by Snail-expression fibroblasts via transforming growth factor β/nuclear factor-κB signaling pathways. CONCLUSION: Taken together, Snail-expressing 3T3 fibroblasts display CAFs properties that support 5-fluorouracil and paclitaxel chemoresistance in CRC via participation of CCL1 and suggest that inhibition of the Snail-expression fibroblasts in tumor may be a useful strategy to limit chemoresistance.

Relationship among levels of peripheral blood miRNA-21 and TGF-β1,CCL5 in patients with acute myocardial infarction and its significance / 心血管康复医学杂志

Objective:To observe levels of peripheral blood miRNA-21,transforming growth factor β1 (TGF-β1) and CC type chemokine ligand 5 (CCL5) in patients with acute myocardial infarction (AMI),and explore their relation-ship and clinical significance.Methods:A total of 57 AMI patients treated in our hospital were selected as AMI group.Another 50 healthy people undergoing physical examination in our hospital simultaneously were selected as healthy control group.Peripheral blood miRNA-21 expression,levels of CCL5,TGF-β1,creatine kinase (CK),CK isoenzyme MB (CK-MB) and cardiac troponin I (cTnI),and coronary Gensini score were measured in all subjects.Relationship among above indexes was analyzed in AMI patients.Results:Compared with healthy control group,there were significant rise in miRNA-21 expression [ (0.67 ± 0.06) vs.(0.89 ± 0.09)],levels of CCL5 [ (17.53 ± 3.75) ng/ml vs.(35.37 ± 5.06) ng/ml],CK [ (85.24 ± 18.33) U/L vs.(173.29 ± 32.56) U/L],CK-MB [ (16.37 ± 2.03) U/L vs.(23.34 ± 2.51) U/L],cTnI [ (11.25 ± 3.23) ng/ml vs.(31.45 ± 8.79) ng/ml] and Gensini score [ (1.65 ± 0.26) scores vs.(43.25 ± 12.13) scores],and significant reduction in TGF-β1 level [ (31.45 ± 8.79) ng/ml vs.(11.25 ± 3.23) ng/ml] in AMI group,P＝0.001 all;in AMI group,peripheral blood miRNA-21 expression was significant positively correlated with levels of CCL5,CK,CK-MB and cTnI (r＝0.476～0.593,P＜0.05 all),and significant inversely correlated with TGF-β1 level and coronary Gensini score (r＝ -0.417,-0.625,P＜0.05 both).Conclusion:Peripheral blood miRNA-21 expression is high,and closely associated with levels of CCL5,TGF-β1 and coronary stenotic degree in AMI patients.It may be used as a potential target for clinical diagnosis and treat-ment of AMI.

The identification of human microRNA335′s predicated target gene CCL1 1,CCL26 and SOX4 / 重庆医学

Objective To identify the targeted-regulating relationship between human MicroRNA335 (hsa-miR-335 )and CCL1 1,CCL26 and SOX4.Methods The potential fragments of hsa-miR-335 target genes CCL1 1,CCL26 and SOX4 were predicted by the bioinformatics analyzing tools online.The 3′untranslated regions(3′UTR)of the CCL1 1,CCL26 and SOX4 were connected to the eukaryotic expression vectors pMIR REPORT.The constructs of pMIR-REPORT-CCL1 13′UTR,pMIR-REPORT-CCL26 3′UTR,pMIR-REPORT-SOX4 3′UTR and positive control were co-transfected with Pre-miRTM miRNA335 Precursor or negative control into 293 T7/1 7 cell line by lipofectamine 2000,respectively.Both Firefly and Renilla luciferase activity were detected by dual luciferase reporter assay system.Results Compared with the negative control group,luciferase assay revealed that has-miR-335 could significantly diminish luciferase activity from SOX4 reporter vector (P 0.05).Conclusion The results suggested that hsa-miR-335 targeted regu-lated SOX4,but not targeted CCL1 1 and CCL26.

CCL1, Specific Manifestation in Lesional Atopic Skin / 대한피부과학회지

BACKGROUND: Chemokines represent a superfamily of small cytokine-like chemoattractive proteins, which regulate leukocyte trafficking under homeostatic and inflammatory conditions. There have been previous reports that chemokine (C-C motif) ligand 1 (CCL1), a member of the family of chemoattractive proteins, is increasingly expressed in atopic dermatitis. OBJECTIVE: We aimed to evaluate the quantity and the pattern of CCL1 expression in atopic dermatitis (AD). METHODS: Biopsy specimens were taken from atopic skin and normal-appearing skin of AD patients as well as psoriatic skin of psoriasis patients. Quantitative real-time PCR analyses and immunohistochemistry of CCL1 expression were performed, and the quantity of CCL1 expression in acute AD was compared with those of normal-appearing atopic skin and psoriatic skin. The serum level of CCL1 was measured by ELISA. RESULTS: CCL1 was most often expressed in acute atopic skin lesions, and the absolute amount of CCL1 18s rRNA in lesional atopic skin was 14.5-fold higher than that in non-lesional atopic skin. Moreover, CCL1 was expressed within the basal layer of the epidermis as well as in the dermis of the lesional atopic skin. However, CCL1 was expressed mostly in the dermis. CONCLUSION: Therefore, CCL1 represents a chemokine that is associated with flare-up of AD, and it may play an important role as a trigger of AD in the initiation and amplification of atopic skin inflammation in the acute phase.

Identification of CCL1 as a Gene Differentially Expressed in CD4+ T Cells Expressing TIM-3

BACKGROUND: T cell immunoglobulin mucin containing molecule (TIM)-3 is expressed in differentiated Th1 cells and is involved in the suppression of the cytokine production by these cells. However, the regulation of the expression of other T cell genes by TIM-3 is unclear. Herein, we attempted to identify differentially expressed genes in cells abundantly expressing TIM-3 compared to cells with low expression of TIM-3. METHODS: TIM-3 overexpressing cell clones were established by transfection of Jurkat T cells with TIM-3 expression vector. For screening of differentially expressed genes, gene fishing technology based on reverse transcription-polymerase chain reaction (RT-PCR) using an annealing control primer system was used. The selected candidate genes were validated by semi quantitative and real-time RT-PCR. RESULTS: The transcription of TIMP-1, IFITM1, PAR3 and CCL1 was different between TIM-3 overexpressing cells and control cells. However, only CCL1 transcription was significantly different in cells transiently transfected with TIM3 expression vector compared with control cells. CCL1 transcription was increased in primary human CD4+ T cells abundantly expressing TIM-3 but not in cells with low expression of TIM-3. CONCLUSION: CCL1 was identified as a differentially transcribed gene in TIM-3-expressing CD4+ T cells.