Nobiletin and its derivatives overcome multidrug resistance (MDR) in cancer: total synthesis and discovery of potent MDR reversal agents

Our recent studies demonstrated that the natural product nobiletin (NOB) served as a promising multidrug resistance (MDR) reversal agent and improved the effectiveness of cancer chemotherapy in vitro. However, low aqueous solubility and difficulty in total synthesis limited its application as a therapeutic agent. To tackle these challenges, NOB was synthesized in a high yield by a concise route of six steps and fourteen derivatives were synthesized with remarkable solubility and efficacy. All the compounds showed improved sensitivity to paclitaxel (PTX) in P-glycoprotein (P-gp) overexpressing MDR cancer cells. Among them, compound 29d exhibited water solubility 280-fold higher than NOB. A drug-resistance A549/T xenograft model showed that 29d, at a dose of 50 mg/kg co-administered with PTX (15 mg/kg), inhibited tumor growth more effective than NOB and remarkably increased PTX concentration in the tumors via P-gp inhibition. Moreover, Western blot experiments revealed that 29d inhibited expression of NRF2, phosphorylated ERK and AKT in MDR cancer cells, thus implying 29d of multiple mechanisms to reverse MDR in lung cancer.     

左旋肉碱修饰的壳聚糖-硬脂酸协载槲皮素口服紫杉醇纳米胶束的制备、表征及在体肠循环研究

目的 制备左旋肉碱修饰的壳聚糖-硬脂酸（LC-SA/CS-SA）纳米胶束，包载紫杉醇（PTX）且协载槲皮素，考察胶束特性，并以大鼠在体肠循环评估给药系统对PTX口服吸收的促进作用。方法 将硬脂酸（SA）通过酰胺化反应接枝于壳聚糖（CS），形成共聚物CS-SA；采用核磁共振H谱、红外光谱鉴定产物结构；以PTX为主药，槲皮素为辅药，采用激光粒径分析、Zeta电位分析和HPLC分析分别考察了胶束的粒径、Zeta电位、载药量、包封率；透射电子显微镜观察胶束形貌；芘荧光探针法测定LC-SA/CS-SA胶束的临界胶束浓度（critical micelle concentration，CMC）；透析袋法考察胶束的体外释放行为；大鼠在体肠吸收实验评估载药胶束的促吸收作用。结果 红外与核磁结果表明SA通过酰胺键接枝于CS；协载槲皮素的LC-SA/CS-SA载PTX胶束呈类球形，粒径为（148.3±1.7）nm，多分散系数（PDI）为0.16±0.07，Zeta电位为（24.600±0.167）mV，CMC为14.31 μg/mL；体外释放结果表明，与市售紫杉醇注射剂相比，协载槲皮素的LC-SA/CS-SA载PTX胶束、LC-SA/CS-SA载PTX胶束具有明显缓释效应；大鼠在体肠吸收实验表明，协载槲皮素的LC-SA/CS-SA胶束对载药PTX的胃肠吸收具有显著促进作用。结论 构建的协载槲皮素的LC-SA/CS-SA载PTX胶束性能优良，促进了PTX的大鼠肠吸收，具有增强药物口服吸收潜力。     

载紫杉醇的大黄酸偶联物纳米胶束的制备及初步评价

目的制备载紫杉醇的D-α-生育酚聚乙二醇1000琥珀酸酯(D-α-tocopheryl polyethylene glycol 1000 succinate,TPGS)修饰的羧甲基壳聚糖-大黄酸偶联物(PTX/TPGS-CR)纳米胶束,并对其进行初步评价。方法采用透析法,以载药量、包封率及粒径为指标,通过单因素考察优化PTX/TPGS-CR纳米胶束的制备工艺并进行验证。以溶血实验及血管刺激性实验初步考察PTX/TPGS-CR纳米胶束的安全性。四甲基偶氮唑盐微量酶反应比色法(MTT)法考察PTX/TPGS-CR纳米胶束对Hela细胞的毒性。通过激光扫描共聚焦显微镜定性和流式细胞仪定量考察Hela细胞对PTX/TPGS-CR纳米胶束的摄取情况。结果制备工艺优化后制得的PTX/TPGS-CR纳米胶束粒径为(197.3±4.4)nm,PDI为(0.131±0.021),电位为(-31.8±0.5)mV,载药量为(48.20±3.03)%,包封率为(87.26±4.91)%。溶血实验结果表明,其溶血率低于1.71%;血管静脉注射无明显刺激性。其对Hela细胞的杀伤作用具有浓度和时间依赖性,能被Hela细胞高效摄取。结论PTX/TPGS-CR纳米胶束载药量和包封率高,安全性好,其体外抗肿瘤活性稍优于Taxol?。     

Multidisciplinary treatment strategy for locally advanced gastric cancer: A systematic review

BackgroundMultidisciplinary management of patients with locally advanced gastric cancer (LAGC) remains unstandardized worldwide. We performed a systemic review to summarize the advancements, regional differences, and current recommended multidisciplinary treatment strategies for LAGC.MethodsEligible studies were identified through a comprehensive search of PubMed, Web of Science, Cochrane Library databases and Embase. Phase 3 randomized controlled trials which investigated survival of patients with LAGC who underwent gastrectomy with pre-/perioperative, postoperative chemotherapy, or chemoradiotherapy were included.ResultsIn total, we identified 11 studies of pre-/perioperative chemotherapy, 38 of postoperative chemotherapy, and 14 of chemoradiotherapy. In Europe and the USA, the current standard of care is perioperative chemotherapy for patients with LAGC using the regimen of 5-FU, folinic acid, oxaliplatin and docetaxel (FLOT). In Eastern Asia, upfront gastrectomy and postoperative chemotherapy is commonly used. The S-1 monotherapy or a regimen of capecitabine and oxaliplatin (CapOx) are used for patients with stage II disease, and the CapOx regimen or the S-1 plus docetaxel regimen are recommended for those with stage III Gastric cancer (GC). The addition of postoperative radiotherapy to peri- or postoperative chemotherapy is currently not recommended. Additionally, clinical trials testing targeted therapy and immunotherapy are increasingly performed worldwide.ConclusionsRecent clinical trials showed a survival benefit of peri-over postoperative chemotherapy and chemoradiotherapy. As such, this strategy may have a potential as a global standard for patients with LAGC. Outcome of the ongoing clinical trials is expected to establish the global standard of multidisciplinary treatment strategy in patients with LAGC.     

lncRNA 178030.2通过TRPS1对三阴性乳腺癌细胞紫杉醇耐药的影响与机制研究

目的:探讨lncRNA 178030.2通过TRPS1对三阴性乳腺癌细胞紫杉醇耐药的影响与机制。方法:采用逐步增加剂量间歇作用的方法诱导三阴性乳腺癌紫杉醇耐药细胞系并命名为MDA-MB-231/R。采用定量PCR和Western blot检测耐药细胞和亲本细胞中lncRNA 178030.2和TRPS1的表达；应用Lipofectamine 2000将lncRNA 178030.2高表达质粒pcmv-178030.2和对照质粒pcmv转染至MDA-MB-231细胞，分别为高表达组和对照组，定量PCR检测lncRNA 178030.2水平的变化，分别用定量PCR和Western blot检测TRPS1 mRNA及蛋白水平的变化；RIP实验检测lncRNA 178030.2是否与TRPS1相结合；MTT法检测MDA-MB-231细胞对紫杉醇的敏感性及细胞增殖。应用Lipofectamine 2000将lncRNA 178030.2小干扰RNA si178030.2和对照siNC转染至MDA-MB-231/R细胞，分别为干扰组和对照组，定量PCR检测lncRNA 178030.2水平的变化，分别用定量PCR和Western blot检测TRPS1 mRNA及蛋白水平的变化；MTT法检测MDA-MB-231/R细胞对紫杉醇的敏感性及细胞增殖。应用Lipofectamine 2000将TRPS1过表达质粒pcmv-TRPS1和对照质粒pcmv转染至MDA-MB-231细胞，分别为高表达组和对照组，分别用定量PCR和Western blot检测两组细胞中TRPS1 mRNA及蛋白水平的表达情况，然后再分别应用Lipofectamine 2000将lncRNA 178030.2高表达质粒pcmv-178030.2转染入两组细胞，MTT法检测MDA-MB-231细胞对紫杉醇的敏感性及细胞增殖。结果:成功构建在3 μg/ml紫杉醇中稳定生长的三阴性乳腺癌耐药细胞系MDA-MB-231/R。定量PCR结果显示:lncRNA 178030.2在紫杉醇耐药细胞系MDA-MB-231/R中的表达明显高于在其亲本细胞MDA-MB-231中的表达；定量PCR和Western blot显示:TRPS1 mRNA和蛋白在紫杉醇耐药细胞系MDA-MB-231/R中的表达明显低于在其亲本细胞系MDA-MB-231中的表达；与对照组相比，高表达lncRNA 178030.2组的MDA-MB-231细胞中TRPS1表达下降，细胞对紫杉醇的敏感性降低，细胞增殖增强，且lncRNA 178030.2确实可以与TRPS1相结合；与对照组相比，低表达lncRNA 178030.2组的MDA-MB-231/R细胞中TRPS1表达升高，细胞对紫杉醇的敏感性升高，细胞增殖减弱；在MDA-MB-231细胞中过表达TRPS1后再过表达lncRNA 178030.2，其促进紫杉醇耐药、促进细胞增殖的作用也明显减弱。结论:lncRNA 178030.2促进三阴性乳腺癌细胞MDA-MB-231的紫杉醇耐药，促进细胞增殖，其发生机制可能与下调TRPS1的表达有关。     

Stathmin and phospho-stathmin protein signature is associated with survival outcomes of breast cancer patients

Currently, Stathmin1 (STMN1) and phospho-STMN1 levels in breast cancers and their clinical implications are unknown. We examined the expression of STMN1 and its serine phospho-site (Ser16, Ser25, Ser38, and Ser63) status by immunohistochemistry. Using Cox regression analysis, a STMN1 expression signature and phosphorylation profile plus clinicopathological characteristics (STMN1-E/P/C) was developed in the training set (n = 204) and applied to the validation set (n = 106). This tool enabled us to separate breast cancer patients into high- and low-risk groups with significantly different disease-free survival (DFS) rates (P < 0.001). Importantly, this STMN1-E/P/C model had a greater prognostic value than the traditional TNM classifier, especially in luminal subtype breast cancer (P = 0.002). Further analysis showed that patients in the low-risk group would benefit more from adjuvant paclitaxel-based chemotherapy (P = 0.002). In conclusion, the STMN1-E/P/C signature is a reliable prognostic indicator for luminal subtype breast cancer and may predict the therapeutic response to paclitaxel-based treatments, potentially facilitating individualized management.     

康莱特注射液联合紫杉醇治疗晚期恶性胸腺瘤的疗效观察

目的:比较康莱特注射液联合紫杉醇及紫杉醇单纯化疗治疗晚期恶性胸腺瘤的不良反应和疗效。方法:2013年8月至2017年12月，经病理学和免疫组化确诊为恶性胸腺瘤并在我院肿瘤中心进行姑息性化疗的患者68例。康莱特注射液联合化疗为观察组（37例）；单纯化疗为对照组（31例）。28天为一周期，共化疗4周期。结果:两组患者均无人发生过敏反应。观察组可有效减轻患者化疗不良反应:恶性、呕吐、四肢麻木、关节肌肉酸痛和白细胞下降(P＜0.05)，可以有效减轻患者胸闷症状和全身疼痛(P＜0.05)，在肿瘤控制方面，康莱特联合化疗的疗效较单纯化疗稍好，但均未见有统计学差异(P＞0.05)。结论:康莱特注射液联合紫杉醇治疗晚期胸腺瘤，具有减轻患者疼痛、减轻化疗不良反应，从而提高患者化疗依从性的作用，值得临床进一步推广和研究。     

红景天苷下调KRT17增加紫杉醇对卵巢癌细胞的化疗敏感性

[目的] 红景天苷是从景天科植物大株红景天的干燥根及根茎或干燥全草中提取的一种化合物，具有多种药理作用。研究旨在探讨红景天苷下调角蛋白家族成员17（KRT17）影响紫杉醇对卵巢癌细胞的化疗敏感性。[方法] 红景天苷、紫杉醇单独或联用处理SKOV-3细胞，将细胞随机分为4组：对照组、红景天苷单独处理组、紫杉醇单独处理组和联合处理组，Cell Counting Kit-8（CCK-8）法检测细胞毒性，5-乙炔基-2’-脱氧尿苷（EDU）检测细胞增殖，Hoechst 33258检测细胞凋亡。免疫荧光法检测自噬体形成情况，蛋白免疫印迹法检测KRT17和细胞自噬相关蛋白表达水平。[结果] 与对照组比较，红景天苷、紫杉醇单独处理组随浓度的升高，对SKOV-3细胞的毒性增强，细胞增殖数、P62蛋白表达降低，细胞凋亡率、LC3荧光斑点数、LC3-Ⅱ/LC3-I比值、Beclin1蛋白表达升高。且红景天苷与紫杉醇协同处理效果优于红景天苷、紫杉醇单独处理组，可能是红景天苷通过下调KRT17蛋白表达实现的。[结论] 红景天苷下调KRT17增加紫杉醇对卵巢癌细胞的化疗敏感性。     

Gemcitabine,paclitaxel, and oxaliplatin (GEMPOX) in the treatment of relapsed/refractory intracranial nongerminomatous germ cell tumors

Intracranial germ cell tumors (GCT) account for less than 5% of all central nervous system tumors in children in Western countries. Approximately 40% are nongerminomatous GCT (NGGCT). Despite correct treatment, 16% to 47% of the patients will relapse. There are no standard approaches in case of recurrence, and treatment in this situation remains a challenge. We report three patients diagnosed with relapsed intracranial NGGCT treated with gemcitabine, paclitaxel, and oxaliplatin, in whom the tumor showed a remarkable response with normalization of tumor markers.