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Objective:To investigate the influence of IL-24 on monocyte activity in patients with non-small cell lung cancer (NSCLC) in vitro. Methods:Twenty-five NSCLC patients and 20 healthy controls were enrolled in the current study. Peripheral blood mononuclear cells (PBMCs) and bronchoalveolar lavage fluid (BALF) samples were collected to isolate monocytes. Expression of IL-22R1, IL-20R1 and IL-20R2 at mRNA level in monocytes was semi-quantified by real-time RT-PCR. Flow cytometry was used to detect the expression of Fas ligand (FasL) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and enzyme linked immunosorbent assay was performed to measure the levels of granzyme A, granzyme B and granzyme H after stimulating monocytes with recombinant human IL-24 (10 ng/ml and 100 ng/ml). The monocytes isolated from peripheral blood of NSCLC patients were co-cultured with A549 cells in vitro. Monocyte-induced target cell death in response to IL-24 stimulation was investigated and changes in the cytotoxicity of monocytes were also assessed after inhibiting granzyme B with Z-AAD-CMK. Student′s t test or LSD- t test was used for comparison. Results:IL-22R1 mRNA was not detected in monocytes. There were no remarkable differences in either IL-20R1 mRNA or IL-20R2 mRNA expression in monocytes between healthy controls and NSCLC patients, or between non-tumor site and tumor site ( P>0.05). FasL and TRAIL expression and granzyme secretion were significantly reduced in monocytes from peripheral blood and tumor sites of NSCLC patients as compared with those from controls ( P<0.05). IL-24 stimulation did not affect the expression of FasL or TRAIL or the secretion of granzyme A or granzyme H ( P>0.05). Low concentration of IL-24 (10 ng/ml) did not affect granzyme B secretion ( P>0.05), while high concentration of IL-24 (100 ng/ml) significantly elevated the secretion of granzyme B by monocytes ( P<0.05). Low concentration of IL-24 (10 ng/ml) did not affect the monocyte-induced target cell death ( P>0.05), while high concentration of IL-24 (100 ng/ml) promoted NSCLC patient-derived monocyte-induced target cell death ( P<0.05). Z-AAD-CMK stimulation suppressed the high concentration of IL-24-mediated elevation of monocyte cytolytic function ( P<0.05). Conclusions:High concentration of IL-24 promoted the cytolytic function of monocytes in NSCLC patients through elevating granzyme B secretion in vitro. However, IL-24 might not influence monocyte function in vivo.
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Objective:To investigate the in vitro influence of IL-24 on the functions of CD8 + T cells from patients with non-small cell lung cancer (NSCLC). Methods:Twenty-eight NSCLC patients and 17 healthy individuals were enrolled in this study. Peripheral blood mononuclear cells (PBMC) and bronchoalveolar lavage fluid (BALF) were collected to isolate CD8 + T cells. Real-time reverse transcription-PCR was used to detect the expression of IL-24 receptors (IL-20R1, IL-20R2 and IL-22R1) at mRNA level in CD8 + T cells. Changes in the expression of perforin and granzyme B were measured by flow cytometry after stimulating purified CD8 + T cells with different concentrations of recombinant human IL-24 (10 ng/ml and 100 ng/ml). In vitro direct and indirect contact co-culture systems were established for CD8 + T cells and NSCLC cell line (NCI-H1882 cells). CD8 + T cells induced target cell death and expression of IFN-γ and TNF-α in response to IL-24 stimulation were analyzed. Student′s t test or LSD- t test was used for intergroup comparison. Results:The expression of IL-22R1 at mRNA level was not detected in CD8 + T cells. No significant difference in IL-20R1 or IL-20R2 expression at mRNA level in CD8 + T cells was observed between healthy individuals and NSCLC patients, or between non-tumor sites and tumor sites ( P>0.05). Perforin and granzyme B expression was significantly reduced in CD8 + T cells from peripheral bloods and tumor sites of NSCLC patients as compared with those from healthy individuals and non-tumor sites (all P<0.05). Low concentration of IL-24 (10 ng/ml) did not affect perforin or granzyme B expression in CD8 + T cells ( P>0.05), but high concentration of IL-24 (100 ng/ml) significantly enhanced the expression of perforin and granzyme B in CD8 + T cells from NSCLC patients ( P<0.05). In the direct contact co-culture system, increased ratio of dead target cells and up-regulated IFN-γ and TNF-α expression were induced after stimulating CD8 + T cells from tumor sites in NSCLC patients with high concentration of IL-24 (100 ng/ml), but low concentration of IL-24 (10ng/ml) had no significant influence on CD8 + T cell-induced target cell death and cytokine production. In the indirect contact co-culture system, neither target cell death nor cytokine production induced by CD8 + T cells was affected by IL-24 stimulation. Conclusions:High concentration of IL-24 promoted the in vitro cytolytic function of CD8 + T cells from NSCLC patients, but might not influence the in vivo functions of CD8 + T cells.
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Objective To investigate the expression of IL-24 in patients with non-small cell lung cancer (NSCLC),and to evaluate its influence on the bioactivity of NSCLC cells. Methods Thirty-nine patient with NSCLC (23 patients with adenocarcinoma and 16 patients with squamous carcinoma) and 17 healthy subjects were enrolled in this study. Serum samples and lung cancer tissues were collected. IL-24 expression in the serum samples was measured using enzyme-linked immunosorbent assay. Its expression at mRNA level in the lung cancer tissues was measured using reverse transcriptional real-time PCR. Adenocar-cinoma cell line A549 and squamous carcinoma cell line NCI-H520 were stimulated with recombinant human IL-24 (10 ng/ml and 100 ng/ml) for 24 hours. Cell proliferation was measured using CCK-8 method. Ap-optosis and cell cycle were measured using flow cytometry. Cell invasion was measured using Transwell as-say. Results Serum IL-24 was significantly elevated in patients with NSCLC in comparison with that in healthy subjects [(144.10±64.43) vs(48.47±18.00) pg/ml]. No significant difference in IL-24 expres-sion was found between patients with adenocarcinoma and squamous carcinoma. IL-24 expression at mRNA level in lung cancer tissues of patients with NSCLC was also significantly increased with an approximately 5-fold enhancement in comparison with that in normal lung tissues. Stimulation with low concentration of re-combinant IL-24(10 ng/ml) promoted the proliferation and suppressed the apoptosis of A549 and NCI-H520 cells. In contrast, high concentration of recombinant IL-24 (100 ng/ml) stimulation notably inhibited the proliferation and enhanced the apoptosis of lung cancer cell lines. No remarkable changes in cell cycle of the two kinds of lung cancer cells in response to IL-24 stimulation were observed. Moreover,low concentration of recombinant IL-24 (10 ng/ml) did not affect the invasion of A549 and NCI-H520 cells,while high concen-tration of recombinant IL-24 (100 ng/ml) significantly inhibited the invasion of lung cancer cells. Conclu-sion IL-24 might influence the bioactivity of NSCLC cells in a concentration-dependent manner. High con-centration of IL-24 might counteract the invasion and metastasis of NSCLC,which is important to prevent dis-ease promotion.
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Objective To investigate the effect of blood glucose level on pulmonary diffusion capacity in elderly patients with type 2 diabetes and its clinical significance.Methods Totally 132 older adults with type 2 diabetes were enrolled in this study.According to blood glucose level,the patients were divided into well-controlled group (n =57) and poor controlled group (n =75).Additionally,52 age-matched non-diabetic healthy subjects were selected as control group.Levels of fasting blood glucose (FBG),glycosylated hemoglobin (HbA1c),fasting insulin (FINS) and 2-hour postprandial glucose (2h PG) in the diabetic patients were detected,and homeostasis model assessment of insulin resistance (HOMA IR) was calculated.The patients with type 2 diabetes underwent urinary albumin excretion rate (UAER) detection,fundus examination and nerve conduction velocity test.pulmonary ventilation,diffusion of the lungs for carbon monoxide (DLCO) and DLCO corrected by alveolar volume (DLCO/VA) were examined in all subjects.Results Levels of FBG,2h PG,HbA1c and HOMA-IR were higher in poor-controlled group than in good glycemicwell-controlled group (all P＜0.05).Compared with the control group,body mass index (BMI) was increased in diabetic groups (both P＜0.05).The pulmonary ventilation function in the three groups had no significant differences (P＞0.05).DLCO and DLCO/VA were lower in diabetic groups than in the control group(all P＜0.05).DLCO and DLCO/VA in poor-controlled group were lower than those in well controlled group (both P＜0.05).DLCO and DLCO/VA were lower in patients with microangiopathy score ≥ 2 than those without microangiopathy (both P ＜ 0.05).Multiple liner regression analysis showed that DCLO and DLCO/VA were negatively correlated with HbA1c,HOMA-IR,duration of diabetes and microangiopathy (r=-2.51,-2.35,-2.42,-2.37,-2.41,-2.52,-2.47,-2.36,all P＜0.05).Conclusions The pulmonary diffusion capacity is significantly impaired in elderly patients with type 2 diabetes.Pulmonary diffusion capacity is negatively correlated with the blood glucose level.The lung may be one of the target organs of type 2 diabetes mellitus.