The role of growth factor administration and T-cell recovery after peripheral blood progenitor cell transplantation in the treatment of solid tumors: results from a randomized comparison of G–CSF and GM–CSF

BACKGROUND: Peripheral blood progenitor cell (PBPC) transplantation (PBPCT) combined with post-PBPCT administration of myelopoietic growth factors is a valid therapeutic intervention to rapidly restore hematopoiesis after the delivery of intensive, myeloablative cancer chemotherapy. On the other hand, the best growth factor regimen to potentiate PBPC-mediated immunohematopoietic recovery has yet to be determined. STUDY DESIGN AND METHODS: In a randomized evaluation, the effects produced by post-PBPCT G-CSF and GM-CSF on myeloid/lymphoid recovery and transplant outcome in women with chemosensitive cancer were compared. Thirty-seven ovarian cancer patients and 34 breast cancer patients ranging in age from 24 to 60 years were treated with carboplatin, etoposide, and melphalan (CEM) high-dose chemotherapy and then randomly assigned to receive G-CSF (5 microg/kg subcutaneously) or GM-CSF (5 microg/kg subcutaneously) until Day 13 after PBPCT. Patients were compared in regard to hematopoietic recovery, posttransplant clinical management, and immune recovery. Finally, clinical outcome was estimated as time to progression and overall survival. RESULTS: Hematopoietic recovery and posttransplant clinical management were comparable in both the G-CSF and GM-CSF series. Conversely, significantly higher T-cell counts were observed in G-CSF-treated patients during the early and late posttransplant follow-up. Patients who received G-CSF showed a significantly longer median time to progression. A parallel analysis revealed that patients in whom a higher CD3+ count was recovered had a significantly longer overall survival and time to progression. CONCLUSION: The enhancement of post-PBPCT T-cell recovery observed in G-CSF-treated patients encourages the use of G-CSF to ameliorate immune recovery, which seems to play a role in post-PBPCT control of disease in cancer patients. GM-CSF might be administered to prolong immunosuppression after autologous PBPCT for autoimmune diseases or allogeneic PBPCT.     

奥美沙坦酯对阿霉素肾病大鼠klotho蛋白表达及氧化应激的影响

目的探讨奥美沙坦酯对阿霉素肾病大鼠klotho蛋白(KL)表达和氧化应激的影响。方法阿霉素肾病大鼠随机分为阿霉素肾病组(ADR组)、奥美沙坦酯干预组(OLM组),每组11只,另设10只正常大鼠作为对照组(NC组)。OLM组大鼠给予奥美沙坦酯10mg/(Kg·d)灌胃,ADR组及NC组大鼠给予等量生理盐水灌胃。于第8周时检测大鼠24h尿蛋白、血清白蛋白、总胆固醇、甘油三酯、尿素氮、肌酐、氧化应激指标(SOD、MDA)、KL mRNA及klotho蛋白表达水平。结果与NC组比较,第8周时ADR组与OLM组24h尿蛋白、总胆固醇、甘油三酯、MDA水平明显增加,KL mRNA、klotho蛋白表达、SOD活性降低,差异具有统计学意义(P<0.05);ADR组与OLM组相比,上述改变更加明显,差异具有统计学意义(P<0.05)。结论阿霉素肾病大鼠klotho蛋白的表达降低,奥美沙坦酯可提高阿霉素肾病大鼠肾脏klotho蛋白的表达,抑制氧化应激,延缓肾病进展。     

siRNA对骨肉瘤U2OS细胞株MDM2-mRNA表达的抑制作用

目的 研究载体介导的siRNA（small interfere RNA）技术对骨肉瘤细胞株U20S中MDM2-mRNA表达的抑制作用.方法 依据设计siRNA的原则,以MDM2为靶基因设计并合成有小发夹结构的两条DNA序列,经退火成互补双链,再克隆到载体PGCsilencerTM中构建重组体（PGCsilencerTM-MDM2 siRNA）.实验分转染重组质粒组,转染阴性对照质粒组和只加脂质体的空白组;RT-PCR法观察U20S细胞中MDM2-mRNA表达改变.结果 成功构建发卡样MDM2 siRNA真核表达载体（PGCsilencer TM-MDM2 siRNA）;PGCsilencer TM-MDM2 siRNA转染到U20S细胞后,MDM2-mRNA表达较空白组显著下降（P＜0.05）,转染阴性对照质粒组与空白组无显著差异（P＞0.05）.结论 成功构建PGCsilencer TM-MDM2 siRNA重组体,并能有效抑制U20S细胞中MDM2-mRNA表达.     

Smoking and FGFR2 rs2981582 variant independently modulate male breast cancer survival: A population-based study in Tuscany,Italy

AimMale breast cancer (MBC) is a rare disease and recommendations for its clinical management are often extrapolated from those for female breast cancer, even if breast cancer (BC) has different characteristics in the two sexes. The purpose of this study was to assess the influence of several individual characteristics including clinico-pathological, lifestyle and genetic factors on overall survival (OS) of a relatively large and well characterized population-based series of 166 MBCs enrolled in Tuscany.MethodsWe genotyped MBC cases at BRCA1/2 genes and at 9 candidate BC susceptibility SNPs. Kaplan-Meier method and multivariate Cox regression, adjusted for several individual characteristics were used. To reduce a possible selection bias related to the interval between diagnosis and enrolment of MBC cases into the study, we used the date of blood donation as the date of the start of observation for survival analysis.ResultsOnly smoking habits had a significant effect on OS at 10 years (for current smokers, HR: 3.34; 95% CI 1.45–7.68; p = 0.004), while lymph node status fell short of reaching statistical significance (for pN positive, HR: 2.07; 95% CI 0.93–4.55; p = 0.07). In the same multivariate analysis we found a significantly higher OS in cases with FGFR2 rs2981582 variant in the dominant transmission model (HR: 0.29; 95% CI: 0.13–0.62; p = 0.028). A sensitivity analysis with left truncation showed similar results.ConclusionsOur results may contribute to shed light on factors influencing MBC survival suggesting an important role for cigarette smoking and FGFR2 rs2981582 variant, and provide clues for better patient management.     

Is OSNA better than imprint cytology for intraoperative diagnosis of cancer involvement of axillary sentinel node in breast cancer?

ObjectiveThe study aim was to establish Sensitivity, Specificity, Positive Predictive Value, Negative Predictive Value (NPV), and Accuracy Values of both imprint cytology (IC) and the OSNA assay for intraoperative assessment of axillary sentinel node (SN) cancer involvement in breast cancer. Specifically, we wished to find out if true positive and false negative results of IC were associated to axillary lymphadenectomy (ALND). Also, we addressed a comparative cost analysis between techniques.Methods244 patients treated for breast cancer in the Breast Unit of Hospital Germans Trias i Pujol from 2011 to 2015 were prospectively included. A transversal, consecutive design was applied to assess IC compared to the reference test (OSNA). Inclusion criteria were: T1 and T2 tumors with negative nodes, both clinically and on ultrasound.ResultsSensitivity of IC for macrometastases was 70%. The NPV of IC for macrometastases was 95,75%. Accuracy of IC was 96,12%. In the comparative cost analysis, the release time of results for OSNA doubled that of IC and was associated with an increased cost of € 370.ConclusionsIC has been stated as a good technique for intraoperative cancer involvement SN with high sensitivity and NPV compared to the OSNA assay. It allows keeping the whole node tissue and thus the possibility of improved histopathological evaluation, which can be useful for adjuvant, and offers the advantage of being less time consuming. Cost analysis shows a higher cost for OSNA, which may exceed the benefit of sorting out false negatives from IC.     

沉默Sema6A基因对骨肉瘤细胞U-2OS增殖和转移的影响

目的检测信号素6A(Sema6A)基因在骨肉瘤细胞系中的表达情况,探讨RNA干扰Sema6A基因对骨肉瘤U-2OS细胞增殖、侵袭及迁移的影响。方法实时定量逆转录聚合酶链反应(qRT-PCR)检测骨肉瘤细胞株U-2OS、Saos-2及MG-63中Sema6A的mRNA表达水平。设计并合成针对Sema6A的小干扰RNA(siRNA)3条及阴性对照siRNA,在Lipofectamine RNAi MAX介导下转染U-2OS细胞,通过qRT-PCR检测Sema6A mRNA水平表达及蛋白质印迹法检测Sema6A蛋白水平,进而筛选出转染效率高的siRNA,CCK-8检测各组细胞活力,Transwell细胞侵袭和迁移实验检测各组细胞的侵袭及迁移能力。结果骨肉瘤细胞株中,Sema6A mRNA在U-2OS中表达最高。瞬时转染Sema6A siRNA的U-2OS细胞中Sema6A的mRNA及蛋白水平较阴性对照组均下降,与阴性对照组相比,实验组细胞的增殖、侵袭及迁移能力均增强,差异有统计学意义(P0.05)。结论 RNA干扰沉默Sema6A基因可以增强U-2OS细胞的增殖、侵袭及迁移能力。