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Objective@#To investigate the effects of macrophage colony-stimulating factor (M-CSF) on the polarization and infiltration of M2 macrophages and the invasion and metastasis of tumor cells in ovarian cancer microenvironment. @*Methods@#A co-culture system consisting of ovarian cancer cells (A2780 and SKOV3) and THP-1 derived macrophages was established in vitro. The M-CSF levels in culture medium and M-CSF mRNA levels in cancer cells and macrophages were detected by ELISA and qRT-PCR, respectively. The proportion of CD68+CD163+ M2 macrophages (polarization cells) was determined by flow cytometry. The invasive and metastatic ability of A2780 and SKOV3 cells after co-culturing with M2 macrophages were analyzed using Transwell assay. The expression levels of M-CSF, CD68+, CD163+ and E-cad in paraffin sections of 52 patients with ovarian cancer and 18 patients with benign ovarian tumor were detected by the immunohistochemistry staining, and their correlations and the relationship between M-CSF and clinicopathological features of ovarian cancer patients were analyzed. @*Results@#The M-CSF levels in culture medium of the co-culture group (A2780 and SKOV3 cells co-cultured with M2 macrophages) were significantly higher than that of A2780 and SKOV3 cells alone (t=14.315 and 12.338, P＜0.01). Fluorescence quantitative PCR results showed that the increased M-CSF originated from the secretion of co-cultured ovarian cancer cells (t=29.915 and 36.826, P＜0.01). The proportions of CD68+CD163+ M2 macrophages in the A2780 cells co-cultured with M2 macrophages group and SKOV3 cells co-cultured with M2 macrophages group were (6.14±0.50)% and (7.32±0.67)%, respectively, which were significantly higher than that in the M2 macrophages alone group ([1.82±0.34]%, t=12.289 and 12.711, P＜0.01). Transwell assay showed that the co-culture environment enhanced the invasion of A2780 and SKOV3 cells (24.00±4.81 vs 75.20±6.42, t=11.058; 18.40±2.31 vs 61.60±9.66, t=7.537, P＜0.01). The expression levels of M-CSF in ovarian cancer tissues were positively correlated with the number of CD68+ cells and CD163+ cells (r=0.690 and 0.596, P＜0.01), and negatively with the expression levels of E-cad (r=-0.566, P＜0.01). Moreover, the expression levels of M-CSF and the number of CD68+ cells and CD163+ cells in ovarian cancer tissues were significantly higher than that in benign ovarian tumor tissues, however, the expression levels of E-cad were on the contrary. The expression levels of M-CSF in ovarian cancer tissues were significantly correlated with tumor stage, differentiation and lymphatic node metastasis (χ2=6.240, 6.612 and 4.544, respectively, P＜0.05). @*Conclusion@#The increased expression of M-CSF in ovarian cancer microenvironment may induce the polarization and infiltration of CD68+CD163+ M2 macrophages, and then promote the invasion and metastasis of ovarian cancer cells.
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Objective To investigate the association between the tumor necrosis factor (TNF)-α-308 G＞A polymorphism and risk of non-Hodgkin lymphoma (NHL).Methods PubMed,CNKI and Wanfang dataknowledge service platform were searched to obtain relevant literature by using the following terms:lymphoma and (tumor necrosis factor or TNF) and (polymorphism or SNP or variant or mutation).Overall results were combined by using fixed effect model or random effect model.Publication bias was evaluated by using the Begg funnel plot and Egger test.Results Fifteen eligible studies were identified,including 9 738 NHL cases and 10 854 controls.The results of the overall meta-analysis showed that the TNF-α-308 AA genotype was associated with increased risk of NHL in two genetic models (AA vs.GG:OR =1.55,95 ％ CI 1.30-1.86,I2 =42.4 ％;AA vs.AG+GG:OR =1.53,95 ％ CI 1.27-1.83,I2 =41.8％).Stratified by ethnicity,TNF-α-308 A allele was associated with an increased NHL risk in Caucasians (A vs.G;AA vs.GG;AG vs.GG;AA vs.AG+GG;AA+AG vs.GG) but a decreased risk in Asians (A vs.G;AG vs.GG;AA+AG vs.GG).Conclusion The TNF-o-308 G＞A polymorphism is associated with the risk of NHL.
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Objective To establish docetaxel (Doc) resistant MDA‐MB‐231/Doc model and epirubicin (Epi) resistant MDA‐MB‐231/Epi mode from triple negative breast cancer cell line MDA‐MB‐231 and to explore their biological characteristics .Methods The MDA‐MB‐231/Doc and MDA‐MB‐231/Epi drug‐resistant cell lines were respectively established by gradually increasing Doc or Epi concentrations induction method in 12 months .The biological characteristics of the cell lines were compared by the cell mor‐phological observation ,MTT and flow cytometry ;the real‐time fluorescent quantitative PCR was used to detect multi‐drug resist‐ance gene (MDR1) mRNA expression;the expression of P glycoprotein(P‐gp) ,estrogen receptor (ER) ,progesterone receptor (PR) and human epidermal growth factor receptor 2 (Her‐2) was detected by Western Blot .Results After the 12‐month induction ,the established MDA‐MB‐231/Doc could grow stably in the medium containing 12 nmol/L Doc ,and MDA‐MB‐231/Epi could grow stably in the medium containing 800 nmol/L Epi;in the same drug concentration ,the growth proliferation rate of the drug resistant cell line was significantly higher than that of the parental generation cells ,their drug resistance indexes were 8 .32 times and 64 .93 rimes of parental generation sensitive cells ,moreover which showed the mutual cross drug resistant status .Compared to the parental generation cells ,the cells of stage G1 and G2 in two cell lines were increased and the cells of stage S were decreased ,with the prolon‐gation of drug withdrawal time ,the cell proliferation speed was accelerated .The expression level of MDR1 gene was increased in the two drug‐resistant cell lines ,which were 4 .05 times and 5 .96 times of parental generation cells respectively ,P‐gp protein expression was positive .Compared with the MCF‐7 cell line ,ER ,PR and Her2 expression in the MDA‐MB‐231 cell line was negative and typi‐cal triple negative breast cancer cell line .Conclusion The drug resistance cell lines of MDA‐MB‐231/Doc and MDA‐MB‐231/Epi are successfully established with stable growth and drug resistance .
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<p><b>OBJECTIVE</b>To explore whether docetaxel-resistant cells (MCF-7/Doc) and doxorubicin-resistant cells (MCF-7/ADM) can secrete Exosomes and their potential role in cell-cell drug-resistance transfer.</p><p><b>METHODS</b>Exosomes were extracted from the cell culture supernatants of MCF-7/Doc and MCF-7/ADM cells by fractionation ultracentrifugation, and were identified by transmission electron microscopy and Western blot analysis. GFP-MCF-7/S, a breast cancer parental sensitive cell line stably expressing green fluorescent protein (GFP), was constructed by recombinant lentiviral vector with GFP. Then the resistance experiment of cells and the experiment of resistance transfer by exosomes were designed to observe the phenomenon of cell-to-cell drug-resistance transfer.</p><p><b>RESULTS</b>Similar to the breast cancer parental sensitive cells (MCF-7/S), the breast cancer resistant sublines could secrete exosomes, which exhibited round or elliptic shape ranging from 30 to 100 nm in diameter with intact membrane, and only expressed the protein marker of exosomes, Tsg101, did not express the endoplasmic reticulum marker calnexin. After MCF-7/S, MCF-7/DOC and MCF-7/ADM cells we cocultured with GFP-MCF-7/S cells for 72 h, there were no significant differences in the expression of fluorescence-labeled cells among the four groups. When treated by the drug ADM or DOC for 24 hours, the MCF-7/DOC+GFP-MCF-7/S group was in favor of a significant higher survival rate of fluorescence-labeled cells compared with the MCF-7/S+GFP-MCF-7/S group (65.5% vs. 25.5%, P < 0.001), and so did the MCF-7/ADM+GFP-MCF-7/S group (53.6% vs. 25.4%, P < 0.001). The exosomes extracted from MCF-7/S, MCF-7/DOC and MCF-7/ADM cells were cultured with the GFP-MCF-7/S cells for 48 h. Among these groups, no significant differences in the expression of fluorescence-labeled cells were found. After treated by the drug ADM or DOC for 24 hours, the exosomes extracted from MCF-7/DOC+GFP-MCF-7/S group was associated with a significant higher survival rate of fluorescence-labeled cells compared with the exosomes extracted from MCF-7/S+GFP-MCF-7/S group (59.9% vs. 32.4%, P < 0.001), and so did the exosomes extracted from the MCF-7/ADM)+GFP-MCF-7/S group (58.3% vs. 27.2%, P < 0.001).</p><p><b>CONCLUSION</b>Our results suggest that drug-resistance can be transferred between breast cancer cells, and exosomes are probably the transporter of the drug resistance.</p>