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1.
建立了检测1640培养液中顺铂含量的火焰原子吸收光谱实验方法,并用该方法来检测人白血病细胞K562经10μg/mL顺铂孵育8 h后,细胞外液的顺铂含量.具体方法为:将含有顺铂的培养液加热干燥后用含有释放剂的无机酸溶剂溶解,火焰原子吸收光谱法测定铂的含量.对无机酸和释放剂进行了筛选,确定了最佳测定条件并检测了上述实验条件下细胞外液中的顺铂含量.结果显示,含顺铂的培养液或细胞外液加热干燥后,可用体积分数为1%HNO3溶液完全溶解,在溶解过程中加入质量分数为1%的La(NO3)3,可使铂成分获得良好的释放效果.K562细胞经10μg/mL顺铂孵育8 h可致细胞活性显著下降(P0.05),此时约6.2%顺铂进入胞内.与传统顺铂检测方法相比,文中的制样方法可使样品不经灰化即可有效检测肿瘤细胞对顺铂的吸收量.  相似文献   

2.
目的:研究凋亡调控相关蛋白来了解顺铂耐药成因,同时考察乙烷硒啉(Ethaselen)在K562耐药细胞中逆转顺铂耐药的作用,并初步探讨其作用机制。创新点:首次研究乙烷硒啉在逆转顺铂耐药中的作用,且此作用与乙烷硒啉诱导细胞凋亡相关。方法:通过长时间脉冲诱导得到顺铂耐药K562细胞,并观察耐药细胞形态及倍增时间。采用MTT法考察乙烷硒啉、顺铂及其联用组在不同细胞株间的生长抑制作用。流式细胞术分析细胞凋亡情况以及细胞内活性氧(ROS)水平。最后,通过蛋白质免疫印迹(Western blot)考察凋亡调控相关蛋白水平的变化。结论:脉冲诱导得到的K562耐药细胞对顺铂的耐受性是原K562细胞的5.34倍。形态学观察发现,耐药细胞体积增大,粘附性进一步降低。乙烷硒啉与顺铂联用表现出协同效应。当加入少量的乙烷硒啉(顺铂与乙烷硒啉的摩尔比率为10:1),顺铂作用K562耐药细胞的半抑制浓度(IC50)值可以减少21倍。流式细胞术及Western blot表明,乙烷硒啉能够诱导耐药细胞凋亡。其逆转顺铂耐药主要是通过调控Bcl-2及Bax蛋白比例以及通过提高细胞内活性氧水平引起线粒体通透转运孔道(PTP)蛋白孔道的形成来促使释放细胞色素c,进而引起Caspase凋亡途径。  相似文献   

3.
目的:研究刺五加多糖对体外培养人白血病K562细胞有无增殖抑制和凋亡诱导作用。方法:取培养至对数生长期的K562细胞(密度为5×104/mL和1×106/mL)分别接种于96孔培养板(100μL/孔)及50mL培养瓶(1.5mL/瓶)中,加入不同浓度的刺五加多糖作用24h后,用CCK-8法检测刺五加多糖对K562细胞增殖抑制作用;荧光显微镜检测细胞凋亡。结果:不同浓度刺五加多糖(0.405、0.810、1.620、2.430、3.240mg/mL)作用K562细胞24 h,抑制率分别为16%、27%、48%、50%、55%;荧光显微镜下观察发现培养K562细胞中出现核固缩、凋亡小体。结论:刺五加多糖对体外培养K562细胞生长有明显的抑制作用,可诱导K562细胞凋亡。  相似文献   

4.
[目的]研究三氧化二砷对K562细胞凋亡的诱导.[方法]采用人红白血病细胞株K562细胞常规培养,给不同浓度的三氧化二砷,在不同的时间收获细胞,用台盼蓝排染法,DNA荧光染料Hoechst33342荧光染色法,及碘化丙啶(PI)与Hoechst33342共染计数坏死细胞的PI阳性率等方法,检测其对K562细胞的影响.[结果]三氧化二砷能够诱导K562细胞凋亡.并且呈现浓度依赖性和时间依赖性.[结论]三氧化二砷主要以诱导肿瘤细胞凋亡而表现其毒性作用.  相似文献   

5.
[目的]研究三氧化二砷对人红白血病细胞株K562凋亡作用及凋亡推测办法.[方法]姬姆萨染色,胎盘兰排染法,双苯并咪唑,碘化丙啶排斥法等.[结果]As2O3诱导12h,K562细胞开始出现凋亡.凋亡细胞随药物浓度的增加和作用时间的延长而增加.各种凋亡推测方法利弊存在差异.[结论]建议使用较有效的Hoechst33342法检测凋亡.  相似文献   

6.
砷剂对肿瘤多药耐药细胞株作用机理的研究   总被引:1,自引:0,他引:1  
三氧化二砷(As2O3)是人们观念中的剧毒物砒霜中的有效成分.它既是一种致癌剂又有一定有益的生物学作用.本实验研究证实,砷剂在非细胞毒性剂量下能降低化疗药物阿霉素(ADM)对多耐药(multi drug resistance,MDR)肿瘤细胞株K562/ADM细胞的IC50(细胞抑制率达50%时化疗药物的浓度),表明三氧化二砷具有逆转人红白血病细胞株K562/ADM细胞MDR作用.  相似文献   

7.
1.静息电位静息电位是指细胞未受刺激时,存在于细胞膜内外两侧的电位差.产生的机理是细胞膜内K+浓度高于细胞外.安静状态下膜对K+通透性大,K+顺浓度差向膜外扩散,膜内的蛋白质负离子不能通过膜而被阻止在膜内,结果引起膜外正电荷增多,电位变正;膜内负电荷相对增多,电位变负,产生膜内外电位差.这个电位差阻止K+进一步外流,当促使K+外流浓度差和阻止K+外流的电位差这两种相互对抗的力量相等时,K+外流停止.膜内外电位差便维持在一个稳定的状态,即静息电位.  相似文献   

8.
目的:通过观察大蒜素对体外培养的人上皮性卵巢癌耐顺铂细胞株(SKOV-3/DDP)的超微结构的影响,以期寻找对耐顺铂治疗的上皮性卵巢癌患者的有效治疗药物。方法:细胞培养:体外培养人上皮性卵巢癌耐顺铂细胞株SKOV-3/DDP,细胞生长至对数生长期后,调整细胞密度为5×l04/ml置于200ml的培养瓶中培养。用透射电子显微镜观察大蒜素作用后SKOV-3/DDP细胞的超微结构:大蒜素组用含25μg/ml大蒜素的培养液培养SKOV-3/DDP细胞,空白对照组用不含药的培养液培养SKOV-3/DDP细胞,两组作用24小时后收集细胞,通过透射电子显微镜观察SKOV-3/DDP的超微结构。结果:大蒜素对SKOV-3/DDP细胞超微结构的影响:透射电子显微镜观察未加药物培养的SKOV-3/DDP细胞呈椭圆形,细胞核为卵圆形,未出现凋亡细胞,经过大蒜素作用后的SKOV-3/DDP细胞则出现明显的超微结构改变并具有典型凋亡细胞特征:细胞核固缩,核膜皱缩,胞浆浓缩并出现大小不等的空泡,染色质浓聚,形成凋亡小体。结论:大蒜素可诱导SKOV-3/DDP细胞呈凋亡改变。大蒜素可诱导SKOV-3/DDP细胞凋亡可能是其对人耐顺铂上皮性卵巢癌的抗癌机制之一,为使大蒜素成为治疗耐顺铂上皮性卵巢癌患者的有效药物提供了可靠的体外实验依据。  相似文献   

9.
顺铂是一种二价铂化合物 ,属细胞周期非特异性化疗药物 ,于 70年代应用于临床肿瘤化疗 ,是治疗头颈部及泌尿生殖系统恶性肿瘤的首选药物之一 ,对食道鳞状细胞癌的姑息治疗也有一定价值。但是 ,临床研究和动物实验表明 ,顺铂与其他抗肿瘤药物一样 ,有许多毒副作用 ,其中主要有 :消化道反应、骨髓抑制、肾脏损害、内耳中毒及周围感觉神经末梢病变等 ,其中以肾毒性和耳毒性最为严重 ,本文主要综述顺铂对耳蜗微循环的中毒性作用及防护机制。1 耳蜗的血液供应及其微循环特点耳蜗有丰富复杂的血管 ,稳定的血流供应在维持耳蜗正常功能及内环境中起…  相似文献   

10.
目的:研究离体热处理和顺铂联合热处理对术中回收血红细胞功能的影响及其中混杂的肝肿瘤细胞株(HepG2)的杀灭作用。创新点:(1)采用多种评价指标研究了不同时间离体热处理对术中回收血中混杂的HepG2的杀灭作用及对红细胞的影响,并确定了对红细胞安全且能有效杀灭HepG2的离体热处理时间。(2)从多个角度评价了离体顺铂联合热处理对术中回收血红细胞的影响及对其中混杂的HepG2的杀灭作用,确定了该方案中对红细胞安全且能有效去除HepG2的顺铂浓度。方法:采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)、5-乙炔基-2’脱氧尿嘧啶核苷(EdU)和平板克隆形成评估HepG2的细胞存活率、DNA复制率和克隆形成能力(图1和3);从红细胞渗透脆性、携氧能力(2,3-二磷酸甘油酸(2,3-DPG)、半饱和氧分压(P50))、能量代谢(Na+-K+-ATPase、pH)、膜完整性(游离血红蛋白(Hb)、血清K+和Na+浓度、细胞膜磷脂酰丝氨酸外翻比例)等角度评估红细胞功能(图2和4;表3)。结论:肝肿瘤术中回收血经离体顺铂联合热处理(42°C,50μg/ml)60min后,能有效清除其中混杂的HepG2,但对红细胞无显著影响,值得体内进一步研究顺铂热处理有效应用于肿瘤手术自体血液回输的安全方案。  相似文献   

11.
It has been reported that Ethaselen shows inhibitory effects on thioredoxin reductase (TrxR) activity and human tumor cell growth. In order to find an efficient way to reverse cisplatin resistance, we investigated the reversal effects of Ethaselen on cisplatin resistance in K562/cisplatin (CDDP) cells that were established by pulse-inducing human erythrocyte leukemic cell line K562, which are fivefold more resistant to cisplatin compared to K562 cells. The morphology and growth showed that the adhesion of K562/CDDP further decreased while the cell volume increased. The proliferation of K562/CDDP is strengthened. The antitumor activities in vitro were assessed by MTT (3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and combination index (CI), showing the significant synergic effects of cisplatin and Ethaselen. Focusing on apoptosis, a series of comparisons was made between K562 and K562/CDDP. Cisplatin induced higher reactive oxygen species (ROS) generation in K562 and subsequently induced the formation of mitochondrial permeability transition pores (PTPs). In addition, cisplatin increased the ratio of Bax to Bcl-2 in K562, which can influence the mitochondrial membrane permeability. PTP formation and mitochondrial membrane permeabilization eventually resulted in the release of cytochrome c and activation of the Caspase pathway. However, these effects were not clearly seen in K562/CDDP, which may be the reason for the acquired CDDP resistance. However, Ethaselen can induce a high level of ROS in K562/CDDP by TrxR activity inhibition and increased ratio of Bax to Bcl-2 in K562/CDDP by nuclear factor κB (NF-κB) suppression, which subsequently induces the release of cytochrome c in K562/CDDP. This response is partly responsible for the reversal of the cisplatin resistance in K562/CDDP cells.  相似文献   

12.
稳恒磁场对人白血病细胞K562生长抑制作用研究初报   总被引:5,自引:0,他引:5  
人白血病细胞K5 6 2经3,6 ,9,12 ,2 4 ,36 ,4 8,6 0 ,72h不同时间磁场处理后,利用MTT法检测其生长活性,台盼蓝拒染法进行活细胞计数.结果表明,K5 6 2细胞经9mT稳恒磁场处理0~72h后,从12h起磁场对细胞生长有显著的抑制效应(p <0 .0 5 ) .K5 6 2细胞生长曲线结果与MTT实验具有良好的一致性.  相似文献   

13.
Homoharringtonine (HHT) has currently been used successfully in the treatment of acute and chronic myeloid leukemias and has been shown to induce apoptosis of different types of leukemic cells in vitro. Emerging evidence suggests that angiogenesis may play an important role in hematological malignancies, such as leukemia. How ever, whether HHT can relieve leukemia by anti-angiogenesis is still unknown. We investigated the anti-angiogenesis potential of HHT with the human umbilical vein endothelial cell line (ECV304) and leukemic cell line (K562) in vitro. Cellular proliferation was determined by MTT assay and apoptosis was analyzed by flow cytometry. The mRNA expression of vascular endothelial growth factor (VEGF) was assessed by RT-PCR and VEGF protein production was detected by Western blot. Inhibition of cell proliferation and induction of apoptosis by HHT were discovered in ECV304 cells, and appeared in a dose- and time-dependent manner. Also, treatment with HHT caused down-regulation of VEGF mRNA expression in K562 cells in similar dose- and time-dependent manner and inhibition of VEGF protein production in K562 cells in response to the enhancing concentration of HHT. The results demonstrated that HHT could also induce apoptosis in endothelium and down-regulate VEGF expression in K562 cells. In conclusion, we believe HHT has anti-angiogenesis potential and speculate that HHT might exert its anti-leukemia effects via reduction of angiogenesis.  相似文献   

14.
INTRODUCTION Homoharringtonine (HHT) is a cephalotoxin alkaloid with anti-leukemic activity and had been used successfully in the treatment of acute and chronic myeloid leukemias (O払rien et al., 1995; 1999; Feldman et al., 1992). The principal mecha-nism of action by HHT is the inhibition of protein synthesis in a dose- and time-dependent manner by binding to ribosome and inhibiting polypeptide chain elongation (Tujebajeva et al., 1989; Zhou et al., 1995). HHT had been shown to indu…  相似文献   

15.
研究噻二唑类杂环化合物对人白血病K562癌细胞的体外抗肿瘤活性筛选。方法:利用MTT法间接测定各化合物对肿瘤细胞后的生长情况,并计算出肿瘤细胞的抑制率,选择抑制率较高的化合物进行深入研究,计算其半数抑制浓度(IC50),并考察其浓度-生长率线性关系。结果:大部分化合物均显示了较好的人白血病K562癌细胞抑制活性,化合物1抑制率达92.01%,IC50为7.07×10-5mol/L,浓度-生长率R2=0.986。结论:噻二唑类杂环化合物具有较好的抗肿瘤活性。  相似文献   

16.
17.
目的:将深海来源的16株真菌经发酵培养与活性筛选,获取活性菌株以供筛选药源活性产物。分别经真菌普通培养基和人工海水培养基发酵获得样品,采用MTT法测试抗肿瘤活性,纸片法测试抗真菌活性。结果:菌株中有3株经过普通发酵培养基发酵和4株经过海水培养基发酵的样品在100μg/mL浓度下对K562细胞的抑制率大于60%;抗真菌活性测试中,仅有菌株16-02-1的发酵样品对受试白色念珠菌ATCC 10231和土曲霉W-1均呈现一定的抑制活性。结论:深海来源真菌在不同培养基中发酵获得的样品,抗肿瘤活性各不相同,经过筛选获得高活性菌株为寻找药源活性产物提供了菌株。  相似文献   

18.
Human thioredoxin reductase (TrxR) system is associated with cancer cell growth and anti-apoptosis process. Effects of 1,2-[bis(1,2-benzisoselenazolone-3(2H)-ketone)]ethane (BBSKE), a novel TrxR inhibitor, were investigated on human leukemia cell lines HL-60 and K562. BBSKE treatment induced cell growth inhibition and apoptosis in both cell lines. Apoptosis induced by BBSKE is through Bcl-2/Bax and caspase-3 pathways. Ehrlich's ascites carcinoma-bearing mice were used to investigate the anti-tumor effect of BBSKE in vivo. Tumor-bearing mice treated with BBSKE showed an increase of life span with a comparable effect to cyclophosphamide (CTX). These results suggest a potential usage of BBSKE as a therapeutic agent against non-solid tumors.  相似文献   

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