羟基喜树碱联合依托泊苷对人Tenon囊成纤维细胞凋亡的影响及其机制

背景研究表明,羟基喜树碱（HCPT）和依托泊苷（VP-16）均能诱导人Tenon囊成纤维细胞（HTFs）的凋亡,但这两种药物联合应用对体外培养的HTFs是否有协同作用尚不明确。目的观察HCPT与VP-16联合应用后在促进体外培养的HTFs凋亡方面是否有协同性,并探讨其作用机制。方法人眼Tenon囊成纤维组织取自江苏省人民医院眼库,采用组织块培养法培养和传代HTFs,采用免疫组织化学法检测HTFs中波形蛋白的表达。第4代HTFs接种于96孔板,分别将不同质量浓度的HCPT（1、5、10、50、100mg／L）、VP-16（0．6、2．5、5．0、25．0、50．0mg／L）、HCPT＋VP-16（质量比为2：1,终质量浓度分别为0．80、3．75、7．50、37．50、75．00mg／L）加入细胞培养孔处理24h,用CCK-8法检测各组药物对细胞增生的抑制率。将HTFs分为空白对照组、HCPT（50mg／L）处理组、VP-16（25mg／L）处理组、HCPT＋VP-16（37．5mg／L）处理组,药物处理24h后用流式细胞仪检测各组细胞的凋亡率。提取细胞内总蛋白,采用Westernblot法检测各药物处理组细胞内caspase-3、活化型caspase-3、bax、bcl-2、JNK、p-JNK、Akt、p-Akt的表达水平。结果培养的细胞呈多边形或不规则形,波形蛋白表达阳性。不同质量浓度HCPT、VP-16和HCPT＋VP-16作用24h后,随着药物质量浓度的增加,对HTFs增生的抑制率增加,差异均有统计学意义（HCPT：F=41．34,P=0．00;VP-16：F=62．60,P=0．00;HCPT＋VP-16：F=46．77,P=0．00）,HCPT处理组、VP-16处理组和HCPT＋VP-16处理组药物的半数抑制浓度（IC50）分别为80．99、27．93、19．81mg／L,HCPT与VP-16联合应用的合并指数（cI）为0．399,表明VP-16和HCPT具有强协同作用。空白对照组、HCPT处理组、VP-16处理组和HCPT＋VP-16处理组HTFs的凋亡率分别为（4．87±0．78）％、（11．20±1．94）％、（12．67±1．51）％和（19．77±2．01）％,差异有统计学意义（F=18．23,P〈0．01）,其中HCPT处理组、VP-16处理组和HCPT＋VP-16处理组HTFs凋亡率较空白对照组明显升高,差异均有统计学意义（q’=15．67、16．32、26．88,P〈0．01）。Westernblot法检测结果表明,与空白对照组相比,HCPT处理组、VP-16处理组和HCPT＋VP-16处理组HTFs中活化型easpase．3、bax、p-JNK表达的灰度值明显增加,差异均有统计学意义（P〈0．01）,且HCPT＋VP-16处理组HTFs中上述各因子表达的灰度值较HCPT处理组、VP-16处理组明显增加,差异均有统计学意义（P〈0．01）,而easpase-3、JNK、Akt的表达无明显变化。与空白对照组比较,bcl-2、p-Akt在HCPT处理组、VP-16处理组和HCPT＋VP-16处理组HTFs中的表达量明显下降,HCPT＋VP-16处理组较HCPT处理组、VP-16处理组的表达量明显减少,差异均有统计学意义（P〈0．01）。结论单独使用HCPT、VP-16均能诱导体外培养的HTFs凋亡,其作用均呈剂量依赖性,HCPT与VP-16联合应用有强协同作用。HTFs中p-JNK、bax的表达上调和p-Akt、bcl-2表达的下调参与联合用药引起的协同效应。     

依托泊苷联合泼尼松持续口服治疗老年人非特异性外周T细胞淋巴瘤

目的探讨持续口服依托泊苷联合泼尼松（EtoP）方案与CHOP方案治疗老年人非特异性外周T细胞淋巴瘤（PTCL—U）的疗效和患者不良反应。方法23例经病理确诊的老年PTCL—U患者,采用抽签法随机分为EtoP组（12例）与CHOP组（11例）,采用Log．rank检验和x2检验对两组的疗效及不良反应进行比较。结果EtoP组总有效率（RR）为66．67％（8／12）,CHOP组为63．64％（7／11）,两组差异无统计学意义（x2=0．023,P=0．879）。EtoP组无进展生存期（PFS）为7．55个月,总生存期（OS）为．15．02个月;CHOP组PFS为4．38个月,0S为12．26个月,两组PFS差异有统计学意义（x2=23．000,P=0．011）,0S差异无统计学意义（x2=14．985,P=0．597）。两组主要不良反应为血液学毒性和消化道反应。其中EtoP组无Ⅲ～Ⅳ度不良反应发生;CHOP组Ⅲ～Ⅳ度血液学毒性发生率为36．36％,Ⅲ-Ⅳ度消化道反应发生率为27．27％。结论对于老年人PTCL—U,EtoP方案疗效略优于CHOP方案,木良反应发生率低于CHOP方案,耐受性良好,应用方便且费用低,值得推广。     

顺铂及依托泊苷对神经母细胞瘤细胞中Trk基因家族表达的影响及临床意义

目的神经母细胞瘤(NB)是儿童常见的实体瘤,而三种不同Trk基因的表达变化与NB的预后及转归关系密切;为研究如何优化化疗药物疗效以提高NB患儿的生存率,本文检测了顺铂(DDP)及依托泊苷(VP16)对人NB的SK-N-SH细胞系的杀伤作用及对细胞中Trk基因家族表达的影响。方法通过CCK-8药物诱导细胞毒性实验检测不同浓度梯度的DDP及VP16对SKN-SH细胞的杀伤作用以及半数杀伤浓度(IC50);用qRT-PCR方法检测DDP及VP16在不同浓度下分别作用24 h及48 h后,对SK-N-SH细胞中TrkA、TrkB和TrkC基因表达变化。结果(1)IDDP及VP16药物浓度的增加与其对SK-N-SH细胞的杀伤作用呈线性关系;且48 h比24 h杀伤作用更明显。(2)SK-N-SH细胞中TrkA及TrkC的表达均随着DDP浓度和作用时间的增加而上调,尤其在10μg/mL浓度、24h最明显;而TrkB的表达在24 h时随着DDP浓度增加而上调,48 h时该促进作用不明显;(3)SK-N-SH细胞中TrkA、TrkB及TrkC的表达亦随着VP16浓度和作用时间的增加而上调。结论 DDP及VP16对NB细胞SK-N-SH有明显杀伤作用并对其Trk基因家族的表达有影响。     

TE方案与CE方案治疗复发性小细胞肺癌的对比研究

目的：观察紫杉醇（Taxol）联合足叶乙甙（VP-16）与卡铂（CBP）联合足叶乙甙治疗复发性小细胞肺癌（SCLC）的有效性和安全性.方法：将44例SCLC分为两组,A组20例,用TE方案（Taxol＋VP-16）,B组24例,用CE方案（CBP＋VP-16）.两组的治疗周期均为28天,完成2个周期后评价疗效及毒副作用.结果：TE方案有效率60%（12/20）,CE组有效率25%（6/24）,两组疗效有显著差异（P〈0.05）,两组毒副作用主要表现为骨髓抑制,恶心、呕吐和脱发,TE组有肌肉关节痛.结论：TE方案治疗复发性SCLC的疗效高于CE方案.     

Results with Floxuridine,Actinomycin D,Etoposide, and Vincristine in Gestational Trophoblastic Neoplasias with International Federation of Gynecology and Obstetrics Scores ≥5

Background5‐fluorouracil‐based multiagent chemotherapy has been used as the primary treatment for high‐risk gestational trophoblastic neoplasia (GTN) in China for a few decades. This study aims to assess the efficacy and toxicity of floxuridine, actinomycin D, etoposide, and vincristine (FAEV) as a primary treatment for patients with GTN who had International Federation of Gynecology and Obstetrics (FIGO) scores ≥5.Materials and MethodsA total of 207 patients with GTN who had FIGO scores ≥5 were treated with FAEV as first‐line chemotherapy at Peking Union Medical College Hospital between January 2002 and December 2017. Complete remission (CR), resistance, survival, toxicity, and reproductive outcomes were analyzed.ResultsOf the 207 patients treated with FAEV, 9 (4.3%) required a change of chemotherapy owing to toxicity and 1 (0.5%) died of cerebral hernia 5 weeks after commencing treatment. The remaining 197 patients were assessable to determine the response to FAEV; among them, 168 (85.3%) achieved CR with FAEV and 29 (14.7%) developed resistance to FAEV. The 5‐year overall survival rate of the entire cohort was 97.4%. Grade 3–4 neutropenia, thrombocytopenia, and anemia occurred in 28.4%, 6.8%, and 6.2% of cycles, respectively. No acute toxicity‐related deaths occurred. Five patients developed acute myeloid leukemia 10–50 months after exposure to chemotherapy; another patient developed duodenal cancer 2 years after completing therapy. Sixty‐one patients who preserved fertility wanted to become pregnant; 56 of them conceived.ConclusionThe FAEV regimen is an effective primary treatment for patients with GTN who have FIGO scores ≥5 and has predictable and manageable toxicity.Implications for PracticeThe most commonly used multiagent chemotherapy for high‐risk gestational trophoblastic neoplasia (GTN) is etoposide, methotrexate and actinomycin D/cyclophosphamide and vincristine (EMA/CO) worldwide. However, 5‐fluorouracil‐based multiagent chemotherapy has been used as a primary treatment for high‐risk GTN in China for a few decades. This study evaluated the efficacy and toxicity of floxuridine, actinomycin D, etoposide, and vincristine (FAEV) as a primary treatment for patients with GTN who have International Federation of Gynecology and Obstetrics (FIGO) scores ≥5. The study''s data demonstrated that FAEV as a primary treatment achieved favorable outcomes for patients with FIGO scores ≥5. Toxicities that result from the FAEV regimen are predictable and manageable. The FAEV regimen may provide another option for the treatment of GTN.     

洛铂或顺铂联合依托泊苷治疗广泛期小细胞肺癌的临床观察

目的 探讨依托泊苷联合洛铂化疗方案治疗小细胞肺癌（SCLC）的近期疗效、远期生存及毒副反应。方法 回顾性分析未经抗肿瘤治疗的广泛期SCLC患者85例,根据治疗方案分为两组,即依托泊苷+洛铂（EL）组42例和依托泊苷+顺铂（EP）组43例。EL组给予依托泊苷80 mg／m²,d₁～d₅,洛铂30 mg／m²,d₁;EP组给予依托泊苷80 mg／m²,d₁～d₅,顺铂25 mg／m²,d₁～d₃,21天为1周期。至少完成2个周期化疗以后评估疗效和毒副反应。 结果 EL组和EP组的有效率（RR）分别为59.5％和53.5％（P＞0.05）,疾病控制率（DCR）分别为80.9％和76.7％（P＞0.05）,但EL组的中位无进展生存时间（PFS）为6.5个月,高于EP组的4.5个月（P＜0.05）。EL组血小板减少的发生率高于EP组,恶心呕吐及肝肾功能损害的发生率均低于EP组（P＜0.05）。结论 同EP方案相比,EL方案可延长中位PFS,且消化道不良反应较轻,基本无肝肾毒性。     

p33ING1b增强骨肉瘤细胞U2OS对足叶乙甙的敏感性

背景与目的作为一个新的抑癌基因,ING1与p53有许多相似的生物学功能,如细胞生长抑制、DNA修复、凋亡和化疗敏感性等.本实验目的在于研究p33ING1b对骨肉瘤细胞药物敏感性的影响并探讨其作用机制.方法瞬时转染p33ING1b进入骨肉瘤细胞株U2OS后,用足叶乙甙(etoposide,VP-16)处理24 h,然后采用台盼蓝拒染法计数活细胞数并计算细胞生长抑制率,流式细胞仪、DAPI染色等方法检测细胞凋亡率,Western blot技术检测p53、p21WAF1、MDM2和Bax蛋白的表达.结果台盼蓝染色结果表明,瞬时转染p33ING1b进入骨肉瘤细胞株U2OS后,再用VP-16处理24 h,细胞生长抑制率明显增加[(63.1±5.1)%].流式细胞计数和DAPI染色检测结果表明,细胞凋亡率明显增加(62.7%).Western blot检测结果显示,外源性p33 ING1b高表达明显提高了p53及其下游基因p21WAF1和Bax蛋白的表达水平,转染p33ING1b进入骨肉瘤细胞株U2OS后,再用VP-16处理24 h,p53、p21和Bax蛋白表达增加.在各实验组中,MDM2的蛋白表达没有明显变化.结论p33ING1b能够上调p53蛋白,并上调p53下游因子--p21WAF1和Bax的表达水平,通过p53依赖性凋亡信号通路,提高骨肉瘤细胞U2OS对VP-16的敏感性.     

Complete response to multidisciplinary therapy in a patient with primary gastric choriocarcinoma

Primary gastric choriocarcinoma is a rapidly growing neoplasm with an average survival of several months in untreated patients.Gastrectomy with lymph node dissection followed by chemotherapy is the treatment of choice.Regimens used for gastric adenocarcinoma are usually selected.However,median survival remains less than six months.In this case report,we describe a case of primary gastric choriocarcinoma with a clinical complete response to multidisciplinary treatment including surgery,chemotherapy,and radiofrequency ablation(RFA).The patient was originally referred for general malaise.Esophagogastroduodenoscopy demonstrated a large tumor occupying the fornix,and total gastrectomy with lymph node dissection was performed.Seven days later,multiple liver metastatic recurrences with high serum levels of beta-human chorionic gonadotropin(β-hCG) were recognized.Chemotherapy with a gonadal choriocarcinoma regimen consisting of etoposide,methotrexate,actinomycin D,cyclophosphamide,and vincristine(EMA/CO),was initiated.After three cycles,serum β-hCG decreased markedly and the tumors disappeared.Six months later,multiple lung metastatic recurrences were found.After one cycle of EMA/CO,only one nodule remained.Computed tomography-guided RFA was performed for this oligometastatic tumor.The patient has been alive with no evidence of disease for 10 years after the initial diagnosis.To the best of our knowledge,this patient with recurrent primary gastric choriocarcinoma has achieved the longest survival.The present case is the first report of choriocarcinoma metastatic to the lung successfully treated with RFA.From our retrospective analysis of recurrent or unresectable primary gastric choriocarcinoma,we propose that gonadal choriocarcinoma regimens can be considered as first-line for primary gastric choriocarcinoma.     

siRNA-mediated Silencing of Survivin Inhibits Proliferation and Enhances Etoposide Chemosensitivity in Acute Myeloid Leukemia Cells

Background: Overexpression of survivin, a known inhibitor of apoptosis, is associated with tumor progressionand drug resistance in numerous malignancies, including leukemias. The aim of this study was to investigate theeffect of a specific survivin small interference RNA (siRNA) on proliferation and the sensitivity of HL-60 acutemyeloid leukemia (AML) cells to the chemotherapeutic drug etoposide. Materials and Methods: The cells weretransfected with siRNAs using Lipofectamine™2000 transfection reagent. Relative survivin mRNA and proteinlevels were measured by quantitative real-time PCR and Western blotting, respectively. Trypan blue exclusionassays were performed to monitor tumor cell proliferation after siRNA transfection. The cytotoxic effects ofetoposide and survivin siRNA, alone and in combination, on leukemic cells were determined using MTT assay.Apoptosis was assessed by ELISA cell death assay. Results: Survivin siRNA markedly reduced both mRNA andprotein expression levels in a time-dependent manner, leading to distinct inhibition of cell proliferation andincreased spontaneous apoptosis. Surprisingly, survivin siRNA synergistically increased the cell toxic effects ofetoposide. Moreover, survivin down-regulation significantly enhanced its induction of apoptosis. Conclusions: Ourstudy suggests that down-regulation of survivin by siRNA can trigger apoptosis and overcome drug resistanceof leukemia cells. Therefore, survivin siRNA may be an effective adjuvant in AML chemotherapy