3-Methyladenine potentiates paclitaxel-induced apoptosis and phosphorylation of cyclin-dependent kinase 1 at thr161 in nasopharyngeal carcinoma cell

Background:Nasopharyngeal carcinoma(NPC)exhibits a significant prevalence in the southern regions of China,and paclitaxel(PTX)is frequently employed as a medication for managing advanced NPC.However,drug resistance is typically accompanied by a poor prognosis.Exploring the synergistic potential of combining multiple chemotherapeutic agents may represent a promising avenue for optimizing treatment efficacy.Methods:This study investigated whether 3-Methyladenine(3-MA)could potentiated the effect of PTX and its potential molecular mechanism.Samples were divided into the following categories:Negative control(NC)with the solvent dimethyl sulfoxide(DMSO,0.5%v/v),PTX(400 nM),3-MA(4 mM),and PTX(400 nM)+3-MA(4 mM).The viability of NPC cells was assessed using both the cell counting kit-8(CCK-8)assay and the colony formation assay.Microscopic observation was performed to identify morphological cell changes.Flow cytometry was used to assess cell cycle status,mitochondrial membrane potential(MMP),and apoptotic cells.Western blotting was conducted to quantify the protein expression.Results:3-MA enhanced PTX-specific inhibition of NPC cell proliferation.PTX,either alone or in combination with 3-MA,caused cell cycle halt at the G2/M phase in the majority of NPC cells,and the combination treatment of PTX with 3-MA induced a higher rate of NPC cell death compared to PTX alone.Western blotting results revealed the combination of PTX with 3-MA heightened activation of cyclin-dependent kinase 1(CDK1),a key molecule in shifting cells from mitotic arrest to apoptosis,led to a reduction in Myeloid Cell Leukemia 1(MCL-1)expression and an increase in Poly(ADP-ribose)polymerase(PARP)cleavage.Conclusion:The concurrent administration of PTX with 3-MA effectively enhances PTX’s inhibitory impact on NPC and activates the apoptosis signal regulated by CDK1.

Paclitaxel induces human KOSC3 oral cancer cell apoptosis through caspase pathways

Background:Paclitaxel is a compound derived from Pacific yew bark that induces various cancer cell apoptosis.However,whether it also has anticancer activities in KOSC3 cells,an oral cancer cell line,is unclear.Methods:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,flow cytometry,and western blotting assays were carried out to assess cell viability,subG1 phase of the cell cycle,and apoptosis-related protein expression,respectively.Results:Ourfindings indicate that paclitaxel could inhibit cell viability and increase the expression of apoptotic markers,including plasma membrane blebbing and the cleavage of poly ADP-ribose polymerase in KOSC3 cells.Also,the treatment with paclitaxel remarkably elevated the percentage of the subG1 phase in KOSC3 cells.In addition,treatment with a pan-caspase inhibitor could recover paclitaxel-inhibited cell viability.Moreover,caspase-8,caspase-9,caspase-7,and BH3 interacting domain death agonist(Bid)were activated in paclitaxel-treated KOSC3 cells.Conclusions:Paclitaxel induced apoptosis through caspase cascade in KOSC3 cells.

Hemorrhagic cystitis in gastric cancer after nanoparticle albuminbound paclitaxel:A case report

BACKGROUND The advanced first-line regimen for advanced gastric cancer is based on a combination of fluoropyrimidine and platinum and/or paclitaxel(PTX),forming a two-or three-drug regimen.Compared to conventional PTX,nanoparticle albumin-bound PTX(Nab-PTX)has better therapeutic effects and fewer adverse effects reported in studies.Nab-PTX is a great option for patients presenting with advanced gastric cancer.Herein,we highlight an adverse event(hemorrhagic cystitis)of Nab-PTX in advanced gastric cancer.CASE SUMMARY A 55-year-old male was diagnosed with lymph node metastasis after a laparo-scopic-assisted radical gastrectomy for gastric cancer that was treated by Nab-PTX and S-1(AS).On the 15th day after treatment with AS,he was diagnosed with hemorrhagic cystitis.CONCLUSION Physicians should be aware that hemorrhagic cystitis is a potential adverse event associated with Nab-PTX treatment.

Efficacy and safety of paclitaxel liposome versus paclitaxel in combination with carboplatin in the first-line chemotherapy for ovarian cancer:a multicenter,open-label,non-inferiority,randomized controlled trial

Background:The paclitaxel liposome formulation,encapsulating paclitaxel within a phospholipid bilayer,ad-dresses the insolubility of traditional paclitaxel formulations,thereby reducing toxicity without compromising its antitumor efficacy.Methods:This multicenter,open-label,non-inferiority randomized controlled trial(ChiCTR2000038555)evalu-ates the efficacy and safety of paclitaxel liposome in comparison to the standard regimen of paclitaxel combined with carboplatin(PLC vs.PC)as first-line therapy in patients with epithelial ovarian cancer.Results:An analysis of median progression-free survival(PFS)revealed non-inferior outcomes between 263 pa-tients in the PLC group and 260 patients in the PC group(32.3 vs.29.9 months,hazard ratio[HR],0.89[95%CI,0.64−1.25]),using a non-inferior margin of 1.3.Although the overall incidence of treatment-related adverse events was comparable between groups,the PLC group experienced significantly fewer non-hematologic toxicities than those treated with the PC regimen.Conclusion:The findings affirm the non-inferiority of paclitaxel liposome compared to the combination of pa-clitaxel and carboplatin regarding therapeutic efficacy,with an enhanced safety profile marked by reduced non-hematologic toxicities.

Ganoderma lucidum spore oil enhances the effect of paclitaxel,improves the tolerance to paclitaxel and prolongs the survival in Lewis tumor-bearing mice

Purpose:This study aims to investigate whether Ganoderma lucidum spore oil(GLSO)could enhance the effect of paclitaxel(PTX),improve the tolerance to PTX and prolong the overall survival of Lewis tumor-bearing mice,which has never been reported before.Methods:The tumor,spleen,and thymus were weighed at the end of the experiment.Whole blood was collected for hematological index analysis,and the intact femur was removed to determine the bone marrow nucleated cell count(BMN).The percentage of lymphocytes in the spleen of mice was detected by flow cytometry,the activity of NK cells was detected by LDH assay,and the proliferation index of lymphocytes was determined by CCK-8 assay.The overall and mean survival time and life extension rate were calculated using SPSS software.Results:Our data showed that GLSO could enhance the anti-tumor effect of PTX and prolong the survival of mice.The underlying mechanisms of the above effects might be related to the toxic reduction effect of GLSO by relieving hematotoxicity,myelosuppression and immunosuppression.Specifically,GLSO could increase the number of blood cells and bone marrow cells,alleviate the thymic index,and elevate the number and activity of NK cells in mice treated with PTX.Conclusion:GLSO may enhance the efficacy of PTX by boosting the activity of immune NK cells and prolong survival by counteracting PTX-induced bone marrow alterations and improving hematopoiesis.These findings suggested the promising role of GLSO in combination with PTX to extend the survival and increase the tolerance of patients in clinical chemotherapy of lung cancer.

产紫杉醇(Taxol)内生真菌的生物多样性

主要阐述了产紫杉醇内生真菌的分离和生物多样性的研究状况 ,紫杉醇产生菌的生物多样性意义及微生物发酵法生产紫杉醇的研究进展 ,并结合内生真菌的特点 ,阐述了紫杉醇产生菌的宿主。

PI-3K/Akt抑制剂LY294002对卵巢癌细胞A2780/Taxol多药耐药性的逆转作用

背景与目的:磷脂酰肌醇(phosphatidylinositol 3 kinase,PI-3K)可抑制细胞凋亡,对PI-3K抑制剂的研究可以更好地了解PI-3K的促癌机制,为多种癌症如卵巢癌、乳腺癌等的基因治疗提供线索。本研究目的在于探讨PI-3K/Akt信号通路抑制剂LY294002对卵巢癌耐紫杉醇细胞株A2780/Taxol多药耐药的逆转作用。方法:用LY294002处理A2780/Taxol细胞,流式细胞术检测细胞凋亡,MTT法检测细胞对紫杉醇的药物敏感性,RT-PCR检测MDR1mRNA的表达,Western blot方法分析LY294002作用前后磷酸化Akt及P-gp蛋白的表达。结果:10和50μmol/L LY294002干预A2780/Taxol细胞24h后,A2780/Taxol细胞凋亡率分别为(8.84±1.65)%和(20.78±2.47)%,显著高于未干预细胞的凋亡率(1.25±0.78)%(P<0.05);A2780/Taxol细胞对紫杉醇的半数抑制浓度(IC50)显著降低(P<0.01),相对逆转效率最高可达(78.08±0.37)%;MDR-1mRNA、磷酸化Akt及P-gp蛋白均明显降低。结论:PI-3K/Akt信号通路的激活与卵巢癌细胞多药耐药的产生有关,PI-3K/Akt抑制剂LY294002可逆转卵巢癌细胞A2780/Taxol的多药耐药。

条件培养液对红豆杉细胞Paclitaxel生产的促进作用

在两步法红豆杉 ( Taxus chinensis)细胞悬浮培养体系的生产阶段 ,加入从生长阶段悬浮培养物中制得的条件培养液 ( Conditioned Medium,CM) ,既能促进细胞的生长 ,又能提高紫杉醇 ( paclitaxel)的产率 ;解决了生产培养时 ,细胞生长受抑制的问题 .特别是 ,取自生长 1 2 d的细胞悬浮培养物的 CM按体积分数为2 5%添加到新鲜生产培养基中时 ,可使细胞紫杉醇最高产量达 2 8.5mg/ L,细胞干重达 32 .2 g/ L,分别是对照的 2 .4倍和 2 .2倍 .对 CM中的蔗糖、果糖、NO- 3 和 PO3- 4

Taxol对缺氧培养乳鼠心肌细胞Cx43蛋白表达及分布的影响

目的:观察微管稳定剂Taxol对缺氧导致的培养乳鼠心肌细胞Cx43表达和分布的影响。方法:差速贴壁梯度离心法分离培养乳大鼠心肌细胞,取培养4d细胞缺氧120min,分别以不同浓度的Taxol干预;免疫印迹法检测聚合态微管蛋白含量、心肌细胞Cx43的蛋白表达,免疫荧光染色后激光共聚焦显微镜观察Cx43的分布。结果:正常培养心肌细胞Cx43分布在核膜和细胞的闰盘处,缺氧120min可导致心肌细胞微管解聚,Cx43蛋白表达降低,在心肌细胞间连接处分布规律散失,核膜Cx43分布减弱或消失,而均匀分布在细胞膜上;在低剂量的Taxol作用下,心肌细胞微管解聚状态缓解,Cx43蛋白表达和分布异常明显改善;随着Taxol剂量增加,这种改善作用更明显,呈现剂量依赖性。结论:缺氧引起乳鼠心肌细胞微管解聚,Cx43蛋白表达降低分布紊乱微管稳定剂Taxol可以显著地保护缺氧导致的心肌Cx43异常,具有潜在的抗缺血性心律失常价值。

Productions of Taxol and Related Taxanes by Cell Suspension Cultures of Taxus yunnanensis

A high taxol yield cell line of Taxus yunnanensis Cheng et L. K. Fu keeps a high taxol_producing level after successive subcultures for more than eight years. In this study, eight taxanes were isolated from the suspension cell cultures of this cell line. Based on NMR and MS analyses, and comparison with literature data and standards, their structures were determined to be 2α,5α,10β_triacetoxy_14β_propionyloxy_4(20),11_taxadiene (1), 2α,5α,10β_triacetoxy_14β_(2′_methyl)_butyryloxy_4(20),11_taxadiene (2), 2α,5α,10β_14β_tetra_acetoxy_4 (20),11_taxadiene (3, taxuyunnanine C), 2α,5α,10β_triacetoxy_14β_(2′_methyl_3′_hydroxy)_butyryloxy_4(20),11_taxadiene (4, yunnanxane) and its 3′_epimer (5), baccatin Ⅳ (6), baccatin Ⅲ (7) and taxol (8), respectively. Among those compounds, 3, 5, 6 and 7 were reported to be isolated from the suspension cell cultures of T. yunnanensis for the first time. TLC and HPLC analyses indicated that the chemical constituents of the culture solution were similar to those of cultured cells. Moreover, the highest taxol content of this cell line reached 0.3% and the cell line could be applied for a large_scale culture.

紫杉醇(Paclitaxel)治疗晚期鼻咽癌的临床研究

评价紫杉醇对晚期鼻咽癌的客观疗效及毒副反应。方法 :采用紫杉醇175mg/m2,3h静脉滴注 (每3周1次 )方案共治疗晚期鼻咽癌病人22例 ,所有病人均接受两个疗程以上的化疗。结果 :22例病人均可评价疗效 ,有7例病人达到PR ,有效率为31 .82 %。主要毒副反应为骨髓抑制、恶心和呕吐、肌痛和关节痛、脱发等。大部分病人为Ⅰ～Ⅱ度反应 ,病人耐受良好。经常规预防用药后 ,未观察到有严重的过敏反应。结论 :紫杉醇是一种对晚期鼻咽癌有效的化疗药物 ,该剂量的紫杉醇临床使用较为安全 ,值得进一步的临床研究。

Isolation and Identification of a Taxol-producing Endophytic Fungus Identified from Taxus media

[Objective] The aim was to isolate and identify a taxol-producing endophytic fungus from Taxus media. [Method] 32 strains of endophytic fungi were identified form the inner bark of T. media,and their fermentation products were detected by high performance liquid chromatography (HPLC). [Result] Through the screening,a strain of taxol-producing endophytic fungi M57 was obtained,which could produce 45-50 μg/L of taxol,and M57 was defined as Rhizopus sp. through morphological observation and 18S rDNA sequence analysis. [Conclusion] The finding of Rhizopus sp. M57 provided a promising strain for producing taxol with taxol-producing fungi fermentation process.

Preparation,physicochemical characterization and cyctotoxicity of solid dispersion of paclitaxel and polyvinylpyrrolidone

The objective of this study was to prepare and characterize paclitaxel-polyvinylpyrrolidone （PTX-PVP） solid dispersions with the intention of improving its solubility and dissolution properties. The PTX-PVP solid dispersion systems were prepared by solvent method. The release rate ofpaclitaxel was determined from dissolution studies and the physicochemical properties of solid dispersion were investigated by differential scanning calorimetry （DSC）, powder X-ray diffraction （PXRD） and scanning electron microscopy （SEM）. The cytotoxicities ofpaclitaxel in solid dispersion to the SKOV-3 cells were assayed by a SRB staining method. The results showed that the solubility and dissolution rate of paclitaxel were significantly improved in solid dispersion system compared with that of the pure drug and physical mixture. The results of DSC and PXRD showed that the paclitaxel in solid dispersion was amorphous form. No paclitaxel crystals in the solid dispersions was found during SEM analysis. Cytotoxicity study suggested that the inhibitory rates of PTX-PVP solid dispersion to SKOV-3 cells were higher than that of pure paclitaxel. The solubility and dissolution of paclitaxel were improved by solid dispersion technique. In vitro cytotoxicity of paclitaxel in solid dispersion was higher than that of pure drug.

微管稳定剂Taxol对缺氧心肌细胞Cx43蛋白的影响

目的观察微管稳定剂Taxol对缺氧心肌细胞Cx43表达和分布的影响,探讨其作为新的抗心律失常药物的潜在价值。方法酶解法分离的大鼠心肌细胞缺氧120 min,并以不同浓度的Taxol干预;台盼蓝排斥实验测定细胞存活率;免疫印迹检测Cx43的蛋白表达,免疫荧光染色后激光共聚焦显微镜观察分析Cx43的分布。结果 Taxol在0.1～10nmol·L-1浓度下成剂量依赖性地提高杆状细胞数,促进细胞存活,而100 nmol·L-1～10μmol·L-1浓度下杆状细胞数开始减少;缺氧时心肌细胞Cx43蛋白表达下降,Taxol呈剂量依赖性地改善心肌细胞的Cx43表达,但过量的Taxol则抑制心肌Cx43;正常心肌细胞Cx43主要分布在细胞两端,缺氧时细胞侧边出现Cx43分布。0.1～10nmol·L-1的Tax-ol呈剂量依赖性地抑制侧边Cx43,100 nmol·L-1～10μmol·L-1的Taxol也抑制心肌细胞两端Cx43。结论缺氧导致心肌细胞Cx43表达降低,分布紊乱,低剂量微管稳定剂Tax-ol可以明显地保护缺氧心肌Cx43,具有潜在的抗缺血性心律失常的临床应用价值。

曼地亚红豆杉双黄酮结构表征和Taxol定量

目的：对曼地亚红豆杉黄酮类成分进行研究，对其不同药用部位紫杉醇含量进行测定。方法：采用反复硅胶柱层析进行分离纯化，根据理化性质、光谱特征鉴定其结构。采用HPLC法测定曼地亚红豆杉枝叶、主根、须根三种药用部位中紫杉醇的含量，色谱柱为C18ODS（150mm×4．6mm，5μm），流动相：甲醇-水（3：1），流速0．8ml／min，检测波长228nm。结果：从曼地亚红豆杉中分离得到三个双黄酮类化合物并进行结构鉴定为金松双黄酮（Ⅰ）、银杏黄素（Ⅱ）、7，7″，4＇-Tri—O—Methylamentoflavone（Ⅲ）。曼地亚红豆杉枝叶、主根、须根中紫杉醇含量依次为：0．0308、0．02191、0．01983mg／g。结论：化合物Ⅰ～Ⅲ为首次从该种植物中得到；曼地亚红豆杉中除树皮外的药用部位枝叶的含量最高，主根次之，须根含量最低。

Nuclestemin siRNA对人肺腺癌紫杉醇耐药株A549/Taxol的耐药性逆转的研究

目的:探讨Nuclestemin基因(NS)特异性RNA干扰对人肺腺癌紫杉醇耐药株A549/Taxol细胞耐药性的逆转作用。方法:NS特异性RNAi表达载体转染A549/Taxol细胞。提取肿瘤细胞总RNA,用RT-PCR方法检测NS的表达;利用MTT法检测紫杉醇的半数抑制浓度(IC50)。结果:NS特异性RNAi表达载体转染A549/Taxol细胞后,NS的表达量明显降低,并且对紫杉醇敏感性的相对逆转率达65.3%。结论:Nuclestemin基因siRNA能有效逆转A549/Taxol细胞的耐药性,明显提高耐药细胞对紫杉醇的敏感性。

人鼻咽癌紫杉醇耐药细胞株CNE-1/Taxol的建立及其机制初探

目的建立耐紫杉醇的人鼻咽癌细胞株,并对其生物学特性及耐药机制进行初步探讨。方法采用大剂量冲击和逐步增加剂量相结合的方法,建立耐紫杉醇鼻咽癌细胞株。检测两种细胞的形态差异、生长曲线及倍增时间;用集落形成实验检测它们对紫杉醇、顺铂和长春新碱化疗药的敏感性;使用RT-PCR比较两种细胞β-微管蛋白Ⅲ,鸟苷酸结合蛋白1的表达差异。结果成功建立了耐药指数为8.43的CNE-1/Taxol耐药细胞株,其增殖速度减慢,倍增期延长,体积变小,且对长春新碱低度耐药,耐药指数为1.33;但与亲本细胞株比较,其对顺铂的敏感性明显增加(P<0.01);β-微管蛋白Ⅲ和鸟苷酸结合蛋白1的表达在耐药细胞株中均升高,并与耐药指数呈正相关(p<0.05)。结论CNE-1/Taxol是一株对紫杉醇耐药的细胞株,是研究紫杉醇耐药机制的重要实验模型;CNE-1/Taxol逆向增加了对顺铂的敏感性;β-微管蛋白Ⅲ和鸟苷酸结合蛋白1表达增强可能是鼻咽癌细胞产生获得性耐药的主要机制之一。

Chitosan对红豆杉PAL及Taxol合成的影响

研究了Ｃｈｉｔｏｓａｎ（聚氨基葡糖）对红豆杉悬浮培养细胞ＰＡＬ（苯丙氨酸解氨酶）活性及Ｔａｘｏｌ（紫杉醇）生物合成的影响．种胚来源的红豆杉细胞培养在改良ＭＳ液体培养基中，于培养的第１８ｄ添加不同浓度的Ｃｈｉｔｏｓａｎ．Ｃｈｉｔｏｓａｎ浓度为１００～１０００ｍｇ·Ｌ－１时对红豆杉细胞ＰＡＬ活性有明显的诱导作用，且诱导作用随浓度的增加而增强，在诱导作用下ＰＡＬ活性在１６ｈ达到峰值；Ｃｈｉｔｏｓａｎ浓度在１００ｍｇ·Ｌ－１时对红豆杉细胞的生长基本无抑制作用，增产效果最显著（约为对照组的１０倍），Ｃｈｉｔｏｓａｎ可作为Ｔａｘｏｌ合成的诱导子．

UTP23基因在A2780/Taxol细胞中表达及对紫杉醇化疗耐药的影响

目的探讨上皮性卵巢癌紫杉醇耐药细胞(A2780/Taxol)中UTP23基因表达水平及其对紫杉醇化疗耐药的影响。方法利用Real-Time PCR与Western blot分别从基因与蛋白表达水平检测A2780/Taxol细胞中UTP23的表达水平,通过脂质体在A2780/Taxol细胞中特异性瞬时过表达UTP23基因,利用MTT细胞增殖法观察过表达UTP23基因对A2780/Taxol细胞药物敏感性的影响;通过Real-Time PCR检测A2780/Taxol细胞过表达UTP23后抗凋亡基因Bcl-2蛋白表达水平。结果 A2780/Taxol中UTP23的m RNA表达水平降低,约为A2780细胞的50%,UTP23蛋白表达水平下降,约为A2780细胞的48%(P<0.01);相对于阴性转染组,UTP23基因过表达可显著提高A2780/Taxol细胞中UTP23基因表达水平(P<0.05);相对于阴性转染组,UTP23基因过表达A2780/Taxol细胞对紫杉醇的敏感性显著提高(P<0.01);与阴性转染组相比,过表达UTP23基因后A2780/Taxol细胞Bcl-2基因表达水平显著下降(P<0.05)。结论 UTP23基因在A2780/Taxol细胞中低表达,可能通过促进Bcl-2基因表达,从而参与A2780/Taxol细胞的耐药作用。

紫杉醇(Paclitaxel,紫素)治疗恶性肿瘤Ⅲ期临床研究报告

为了对紫杉醇的临床应用价值和药物不良反应进行进一步评价，根据协作组共同制定的Ⅲ期临床试用计划通过前瞻性多中心１６单位进行临床研究。共收治２４３例恶性肿瘤患者，均为有病理或细胞学证实的中晚期病人。单药治疗：所选病人大多为一般状况较好，首次治疗的晚期患者。应用紫杉醇１５０～１７５理学ｍｇ·ｍ－２，静脉滴注，３～５ｈ，每３～４周一次，２～３周期为一疗程。联合化疗主要为经手术、化疗、放疗后的晚期患者，所用方案为：紫杉醇静脉滴注１３５ｍｇ·ｍ－２，卵巢癌加顺铂８０ｍｇ·ｍ－２；乳腺癌加阿霉素４０ｍｇ·ｍ－２；肺癌加顺铂８０ｍｇ·ｍ－２或静脉滴注卡铂３５０ｍｇ·ｍ－２；食管癌加静脉滴注顺铂８０ｍｇ·ｍ－２，第１周和平阳霉素８ｍｇ，肌注２周，第１、２周使用。均每３周重复一次，２～３周期为一个疗程。结果本组可统计近期疗效的１９０例，治后完全缓解１４例，部分缓解７３例，无变化７７例，进展２６例，总有效率为４５．８％。卵巢癌单药治疗的有效比为３／４，与顺铂联合应用的有效率为３０％（６／２０）；乳腺癌单药治疗为６２．５％（１０／１６），与阿霉素联合应用为６０．０（２４／４０）；食管癌单用有效比为４／５，与顺铂及平阳霉素联合应用为?