miR-874 functions as a tumor suppressor by inhibiting angiogenesis through STAT3/VEGF-A pathway in gastric cancer

MicroRNAs are endogenously expressed, small non-coding RNAs that regulate gene expression by targeting mRNAs for translational repression or degradation. Our previous studies indicated that miR-874 played a suppressive role in gastric cancer (GC) development and progression. However, the role of miR-874 in tumor angiogenesis and the mechanisms underlying its function in GC remained to be clarified. Here, gain- and loss-of-function assays demonstrated that miR-874 inhibited the tumor angiogenesis of GC cells in vitro and in vivo. Through reporter gene and western blot assays, STAT3 was shown to be a direct target of miR-874. Overexpression of STAT3 rescued the loss of tumor angiogenesis caused by miR-874. Conversely, the STAT3-shRNA attenuated the increased tumor angiogenesis caused by the miR-874-inhibitor. Furthermore, the levels of miR-874 were inversely correlated with those of STAT3 protein in GC tissues. Taken together, these findings indicate that down-regulation of miR-874 contributes to tumor angiogenesis through STAT3 in GC, highlighting the potential of miR-874 as a target for human GC therapy.     

Involvement of Ras GTPase-activating protein SH3 domain-binding protein 1 in the epithelial-to-mesenchymal transition-induced metastasis of breast cancer cells via the Smad signaling pathway

In situ models of epithelial-to-mesenchymal transition (EMT)-induced carcinoma develop into metastatic carcinoma, which is associated with drug resistance and disease recurrence in human breast cancer. Ras GTPase-activating protein SH3 domain-binding protein 1 (G3BP1), an essential Ras mediator, has been implicated in cancer development, including cell growth, motility, invasion and apoptosis. Here, we demonstrated that the upregulation of G3BP1 activates the EMT in breast cancer cells. Silencing Smads almost completely blocked this G3BP1-induced EMT, suggesting that this process depends on the Smad signaling pathway. We also found that G3BP1 interacted with the Smad complex. Based on these results, we proposed that G3BP1 might act as a novel co-factor of Smads by regulating their phosphorylation status. Moreover, knockdown of G3BP1 suppressed the mesenchymal phenotype of MDA-MB-231 cells in vitro and suppressed tumor growth and lung metastasis of 4T1 cells in vivo. Our findings identified a novel function of G3BP1 in the progression of breast cancer via activation of the EMT, indicating that G3BP1 might represent a potential therapeutic target for metastatic human breast cancer.     

E2F7 overexpression leads to tamoxifen resistance in breast cancer cells by competing with E2F1 at miR-15a/16 promoter

About 50–70% of breast cancers are estrogen receptor α (ERα) positive and most of them are sensitive to endocrine therapy including tamoxifen. However, one third of these patients will eventually develop resistance and relapse. We found that the expression of miR-15a and miR-16 were significantly decreased in tamoxifen resistant ER positive breast cancer cell lines. Exogenous expression of miR-15a/16 mimics re-sensitized resistant cells to tamoxifen by inhibiting Cyclin E1 and B cell lymphoma-2 (Bcl-2) to induce cell growth arrest and apoptosis respectively. Further, we identified that a repressive member of E2F family, E2F7, was responsible for the suppression of miR-15a/16 cluster by competing with E2F1 for E2F binding site at the promoter of their host gene DLEU2. Moreover, high expression of E2F7 is correlated with high risk of relapse and poor prognosis in breast cancer patients receiving tamoxifen treatment. Together, our results suggest that overexpression of E2F7 represses miR-15a/16 and then increases Cyclin E1 and Bcl-2 that result in tamoxifen resistance. E2F7 may be a valuable prognostic marker and a therapeutic target of tamoxifen resistance in breast cancer.     

Retargeted human avidin-CAR T cells for adoptive immunotherapy of EGFRvIII expressing gliomas and their evaluation via optical imaging

There has been significant progress in the design of chimeric antigen receptors (CAR) for adoptive immunotherapy targeting tumor-associated antigens. However, the challenge of monitoring the therapy in real time has been continually ignored. To address this issue, we developed optical molecular imaging approaches to evaluate a recently reported novel CAR strategy for adoptive immunotherapy against glioma xenografts expressing EGFRvIII. We initially biotinylated a novel anti-EGFRvIII monoclonal antibody (biotin-4G1) to pre-target EGFRvIII⁺ gliomas and then redirect activated avidin-CAR expressing T cells against the pre-targeted biotin-4G1. By optical imaging study and bio-distribution analysis, we confirmed the specificity of pre-target and target and determined the optimal time for T cells adoptive transfer in vivo. The results showed this therapeutic strategy offered efficient therapy effect to EGFRvIII⁺ glioma-bearing mice and implied that optical imaging is a highly useful tool in aiding in the instruction of clinical CAR-T cells adoptive transfer in future.     

EBV-miR-BART10-3p facilitates epithelial-mesenchymal transition and promotes metastasis of nasopharyngeal carcinoma by targeting BTRC

Epstein-Barr virus (EBV) infection is closely associated with tumorigenesis and development of nasopharyngeal carcinoma (NPC), but the underlying molecular mechanisms remain poorly understood. It has been recently reported that EBV encodes 44 mature miRNAs, some of which were found to promote tumor development by targeting virus-infected host genes or self-viral genes. However, few targets of EBV encoded-miRNAs that are related to NPC development have been identified to date. In this study, we revealed that in NPC cells, EBV-miR-BART10-3p directly targets BTRC gene that encodes βTrCP (beta-transducin repeat containing E3 ubiquitin protein ligase). We found that EBV-miR-BART10-3p expression in clinical samples from a cohort of 106 NPC patients negatively correlated with BTRC expression levels. Over-expression of EBV-miR-BART10-3p and down-regulation of BTRC were associated with poor prognosis in NPC patients. EBV-miR-BART10-3p promoted the invasion and migration cabilities of NPC cells through the targeting of BTRC and regulation of the expression of the downstream substrates β-catenin and Snail. As a result, EBV-miR-BART10-3p facilitated epithelial-mesenchymal transition of NPC. Our study presents an unreported mechanism underlying EBV infection in NPC carcinogenesis, and provides a potential novel biomarker for NPC diagnosis, treatment and prognosis.     

体外循环手术中血小板单采对血液的保护作用

目的　探讨在体外循环心脏手术中血小板单采对血液的保护作用 ;方法　选择 2 0例行心脏瓣膜手术患者 ,随机分为实验组和对照组 ,实验组采用血小板单采技术 +常规体外循环 ,对照组只采用常规体外循环 ,其他血液保护措施两组相同 ;结果　①实验组血小板采集量均在患者血小板总数的 2 0 %以上 ;②实验组术中血液制品 (全血和血浆 )用量较对照组明显减少 ,差异有显著性 ;③两组间Hb及Hct各时点变化趋势一致 ;④围手术期患者血小板计数的动态变化两组差异不大 ,而实验组恢复较快 ;⑤实验组术后血小板聚集功能明显优于对照组 ,差异有显著性 ;⑥实验组术后凝血酶原恢复时间明显较对照组快 ,差异有显著性 ;⑦实验组术后各分时段引流量较对照组明显减少 ,差异有显著性。结论　体外循环术中血小板单采能够保护血小板免遭体外循环的破坏 ,明显减少术后出血 ,起到良好的血液保护作用。     

Awake language mapping and 3-Tesla intraoperative MRI-guided volumetric resection for gliomas in language areas

The use of both awake surgery and intraoperative MRI (iMRI) has been reported to optimize the maximal safe resection of gliomas. However, there has been little research into combining these two demanding procedures. We report our unique experience with, and methodology of, awake surgery in a movable iMRI system, and we quantitatively evaluate the contribution of the combination on the extent of resection (EOR) and functional outcome of patients with gliomas involving language areas. From March 2011 to November 2011, 30 consecutive patients who underwent awake surgery with iMRI guidance were prospectively investigated. The EOR was assessed by volumetric analysis. Language assessment was conducted before surgery and 1 week, 1 month, 3 months and 6 months after surgery using the Aphasia Battery of Chinese. Awake language mapping integrated with 3.0 Tesla iMRI was safely performed for all patients. An additional resection was conducted in 11 of 30 patients (36.7%) after iMRI. The median EOR significantly increased from 92.5% (range, 75.1–97.0%) to 100% (range, 92.6–100%) as a result of iMRI (p < 0.01). Gross total resection was achieved in 18 patients (60.0%), and in seven of those patients (23.3%), the gross total resection could be attributed to iMRI. A total of 12 patients (40.0%) suffered from transient language deficits; however, only one (3.3%) patient developed a permanent deficit. This study demonstrates the potential utility of combining awake craniotomy with iMRI; it is safe and reliable to perform awake surgery using a movable iMRI.     

HPLC法测定健脾养胃颗粒中厚朴酚的含量研究

目的:建立健脾养胃颗粒中厚朴酚的含量测定方法。方法:采用HPLC法,色谱柱:采用Allsphere ODS(4.6 mm×250 mm,5μm);流动相:甲醇-0.1%醋酸(78:22),流速:1.0 ml.min-1,检测波长为294 nm。结果:厚朴酚在0.05792~0.5792μg范围内线性良好,r=0.9999,平均回收率99.18%,RSD%=1.80%(n=6)。结论:该方法分离度高,重现性好,简便,准确,可用于健脾养胃颗粒的质量控制。