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The study aimed to elucidate the modulatory role of MMP14 on mCD100 shedding and sCD100 production,and its subsequent effects on CD8+T cell dysfunction in lung cancer patients.Total of 56 non-small cell lung cancer(NSCLC)patients were from January 2020 to January 2023 and compared them with 88 healthy controls.Bronchoalveolar lavage fluid(BALF)was obtained from both tumor and non-tumor sites of the patient group.Peripheral blood mononuclear cells(PBMC)were isolated from both groups,and the expression of CD72 and mCD100 in PBMC were assessed via flow cytometry.CD8+T cells from tumor sites were stimulated with recombinant human MMP14 and CD100.Post-cultivation,supernatant levels of TNF-α and IFN-γ were determined by ELISA,while granulysin B and perforin levels were analyzed through an ELISPOT assay.The rate of target cell death was also observed.Data showed no significant difference in the proportion of CD3+mCD100+,CD3+CD72+ cells,and the average fluorescence intensity of CD72 in CD100 and CD3+ monocytes in CD3+CD8+T cells between the patient and control groups.However,as compared with non-tumor sites,these indexes of tumor sites were significantly elevated.Stimulation with CD100 led to increase in IFN-γ,TNF,perforin,and granulozyme B secretion levels in CD8+T cells.After MMP14 stimulation,the proportions of CD3+mCD10 0+ and target cell death,along with sCD100,TNF-α,IFN-γ,and granulozyme B levels in CD8+T cells from NSCLC tumor sites,were notably increased.Interestingly,the addition of anti-CD100 to MMP14-stimulated CD8+T cells resulted in a significant drop in the levels of sCD100,TNF-α,IL-1β,and granulozyme B,as well as in the proportion of target cell death.Taken together,in NSCLC patients,the inhibition of CD100 shedding in CD8+T cells at tumor sites and the blockade of sCD100 production result in impaired CD8+T cell killing function.MMP14 appears to enhance mCD100 shedding and sCD100 production,thereby potentially restoring the cytotoxic function of CD8+T cells against primary NSCLC cells.
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Objective To investigate tumor cell killing effect of superparamagnetic Fe3O4 nanoparticles with cubic phase through magneto-mechanical force under a low-frequency vibrating magnetic field ( VMF). Methods A kind of strong magnetic and irregular-shaped Fe3O4 nanoparticles with cubic phase was synthesized by coprecipitation method. The Fe3O4 nanoparticles were exposed to a self-developed VMF and cell killing efficiency of the Fe3O4-mediated magneto-mechanical force was investigated. Results VMF alone had no effects on cell viability. After Fe3O4 nanoparticles were added, the cell viability significantly decreased with prolonging the VMF treatment time and increasing the Fe3O4 nanoparticle concentration. Lactate dehydrogenase released by damaged cells also increased with prolonging the VMF exposure time. Conclusions The irregular-shaped Fe3O4 nanoparticles can transfer magneto-mechanical force to tumor cells under VMF, cause structural damage of cells and result in cell death. The VMF generator developed in this study has simple structure and it is safe for use and convenient for operation. The developed magnetic nanoparticles and the corresponding cancer cell killing technique have the potential for clinical transformation.
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@#Introduction: Patients with Nasopharyngeal carcinoma (NPC) usually diagnosed at advanced cancer stage and recurrent case. Rac1 have become an emerging therapeutic target for metastasis cancer. This gene is critically involved in cell polarization and reactive oxygen species-mediated cell killing. This study aims to investigate the Rac1 activities in NPC/HK1 cell line using siRNA approach and evaluate the calcium deposition profile. Methods: The NPC/ HK1cells were transfected with Rac1-siRNA (siRac1) at concentrations of 50nM, 100nM and 200nM for 24 hours and stained with alizarin red s for calcium mineralization profile. Levels of Rac1 gene expression were measured via qRT-PCR followed by the time dependent assessment for 24, 48 and 72 hours. Results: Findings revealed that siRac1 concentrations of 200nM (p-value <0.02) and 100nM (p-value <0.016) had significant Rac1 suppression while 50nM (p-value <0.076) had the least suppression. On the other hand, from alizarin red S staining showed no significant changes for calcium mineralization activity on treated and control cells. However, siRac1 treated cells at 200nM showed presence of intracellular organelle swelling and loss of membrane integrity in 70% of the cells. This observation could possibly be linked to early sign of necrosis activity, hypoxia and disruption in intracellular calcium influx. Conclusion: This study suggest that Rac1 gene suppression might be involved in disruption of calcium deposition and reactive oxygen species-mediated NPC/HK1 cell killing. Further insight on the Rac1 molecular mechanism are needed to understand its potential role as therapeutic target for NPC
Sujet(s)
Cancer du nasopharynxRÉSUMÉ
Objective To investigate the effects of c-Jun N-terminal kinase (JNK) 2 on NKG2D-mediated natural killer ( NK) cell cytotoxicity .Me thods NK cells were activated by polyinosinic-polycyti-dylic acid ( Poly I∶C ) .The activation status of JNK signaling pathway was detected .The cytotoxicity of ac-tivated NK cells and the level of IFN-γproduced were measured to determine the function of NK cells in the absence of JNK2.Tumor growth in wild type and JNK2-knockout (JNK2-/-) mice with lymphoma xenograft were measured to evaluate immune surveillance of NK cells .Results The phosphorylation of JNK and up-stream kinases were observed in the early stage of cell activation after treatment of Poly I ∶C.The expressed ligands of the activating receptors NKG2 D significantly increased NK cell cytotoxicity to lymphoma cells . JNK2 deficiency impaired the antitumor effects of NK cells , and then resulted in enhanced tumor growth in JNK2-/-mice.Conclusion JNK2 signaling pathway is involved in the NKG2 D-mediated activation of NK cells and regulates immune surveillance of NK cells against tumor .
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Objective To study the anti-tumor effects of mixed cultured B16 melanoma cells supernatant.Methods The supernatant from purely cultured B16 melanoma cells or mixedly cultured B16 melanoma cells with lymphocytes and macrophages at indicated time points were collected,respectively.The chemotaxis of the two different cell supernatants was determined by Boyden room method.The activation effects towards lymphocytes of the two different supernatants were determined by CCK-8 method.Results When the cells were mixed cultured for 0.5,1,2,4,8 and 12 h,the numbers of lymphocytes to travel from the upper well to the bottom well were (1.00±0.82) × 104,(7.00±1.63) × 104,(9.50±0.58) × 104,(11.25±2.36) ×104,(17.25±1.71) × 104 and (21.50±1.29) × 104,respectively.When the cells were purely cultured for 0.5,1,2,4,8 and 12 h,the numbers of lymphocytes to travel from the upper well to the bottom well were (0.00±0.00) ×104,(0.25±0.50) × 104,(1.75±0.96) × 104,(5.25±0.96) × 104,(5.75±1.26) × 104 and (10.75±3.20) × 104,respectively.The mixed cultured group showed higher chemotaxis effects towards lymphocytes in comparison with the purely cuhured one at the same points except for 0.5 h (P ＜ 0.05).The mixed cultured group showed higher activation effects towards lymphocytes in comparison with the purely cultured at the same points except for 0.5 and 1 h (P ＜ 0.05).Each group showed higher chemotaxis and activation effects towards lymphocytes when they were cultured for 12 h than the other time points (P ＜0.05).Conclusion The supernatant from mixed cultured cells shows much higher chemotaxis and activation effects towards lymphocytes to kill tumor cells.