氨磷汀对顺铂肾毒性损伤的保护作用及其机制的研究

目的 观察顺铂的肾毒性损伤部位、形式与肾功能检查结果的相关性,以了解细胞凋亡的发生机制和氨磷汀的保护机制是否与肾组织Fas和FasL表达改变有关。方法 随机将大鼠分成3组,即对照组（生理盐水）、顺铂组（6mg／kg）和氨磷汀组（顺铂6mg／kg＋氨磷汀200mg／kg）,取其血清标本和肾组织,分别做血清BUN、Cr检测和肾组织病理学检查,并用原位缺口末端标记法（TUNEL）做肾组织凋亡细胞检测、Fas和FasL免疫组化染色,再用图像分析软件对其做阳性细胞计数和染色总灰度值测定。结果 顺铂组动物血清BUN、Cr值均明显高于对照组和氨磷汀保护组,3d时,差异已有统计学意义（P〈0．05）;5d时,两者差异分别为P〈0．01和P〈0．05;10d时,恢复正常。顺铂组肾小管上皮细胞坏死和凋亡均很严重,其凋亡细胞计数明显高于对照组和氨磷汀组（P值均〈0．01）,肾组织Fas和FasL表达的总灰度值,明显高于对照组和氨磷汀组（P值均〈0．01）。结论 氨磷汀对顺铂的肾毒性损伤有保护作用,其机制可能与抑制肾组织Fas和FasL的表达有关。     

顺铂的肾毒性作用及其防治

顺铂(cisplatin)是一种广谱抗癌药物,但其对肾脏的毒性作用,在一定程度上限制了它的应用。单用顺铂时,肾毒性发生率高达28～36％。近年来许多学者致力于探讨顺铂的肾毒性机理及其防治方法,并取得了一定进展。本文对此作一简要综述。 一、肾毒性及其机理 顺铂的肾毒性作用主要是由金属铂离子产生。血浆中高浓度的铂离子在用药后6小时就可对肾小管产     

顺铂的肾脏毒性及其预防

 顺铂(Cisplatin DDP)是当前最有效的化疗药物之一,但是它也有许多不良反应,常见的有重度消化道反应、耳毒性、肾脏毒性、骨髓抑制、末梢神经病、离子紊乱等,其中的肾脏毒性尤为突出，一直是限制DDP临床应用的主要因素.现就DDP的肾损伤及其预防措施综述如下。     

免疫毒素与顺铂协同杀伤肿瘤细胞机制的探讨

背景与目的:免疫毒素HELβ-PE38KDEL(HeregulinEGFlikedomainβ1-PsedomonasAeurginosa38KDEL,本文简称ITH)已经被证实是一种具有特异性杀伤erbB2、3、4阳性乳腺癌细胞的新的生物学制剂,然而它与化疗药物的联合作用效果目前尚未有报道。本文探讨ITH与化疗药物顺铂(DDP)的协同抗肿瘤作用。方法:采用AnnexinV结合实验和Westernblot检测乳腺癌细胞MDA-MB-453、2LMP和胃癌细胞N87分别在ITH和DDP单独及联合用药前后的细胞凋亡变化。结果:在高表达erbB-2、3、4的MDA-MB-453和N87中,联合用药组和单药组相比,凋亡细胞成倍增加(P<0.01);而在低表达erbB-2、3、4的2LMP,联合用药组和单药组相比,凋亡细胞无明显增加(P>0.05)。MDA-MB-453和N87细胞在联合用药组中PARP、caspase-3降解增加,bcl-2、mp53过度表达受抑制;而2LMP细胞中PARP、caspase-3降解无增加,bcl-2、mp53过度表达未受抑制。结论:ITH和DDP联合后可促进高表达erbB-2、3、4的乳腺癌细胞凋亡,这可能是两者协同作用的机制之一。     

顺铂肾毒性作用的初步实验研究

顺铂(DDP)的肾毒性作用是影响临床应用的关键因素,其机理目前尚不清楚。本文进行了初步动物试验观察,以提供初步参考材料。     

高温合并顺铂对人肺腺癌细胞株H1299的协同杀伤作用

目的:观察高温合并顺铂对人肺腺癌细胞株H1299的协同杀伤作用。方法:用CCK-8法检测高温处理或未处理的细胞对顺铂的敏感性。单独42℃作用2h、顺铂10μg/ml作用24h和两者联合作用后,采用CCK-8法检测细胞抑制率和流式细胞仪分析细胞各时相DNA含量,观察两者对细胞毒性和细胞周期的影响。结果:高温作用后,细胞对顺铂的半数抑制浓度(IC50)从9·51μg/ml降低至6·07μg/ml;高温和顺铂联合作用对细胞有明显的协同杀伤作用,流式细胞检测也显示高温对细胞周期有明显的影响,G0/G1期细胞比例明显增加,S期细胞比例明显下降。结论:高温可以明显提高H1299细胞对顺铂的敏感性。     

顺铂诱导宫颈癌Hela细胞凋亡及其作用机制的研究

目的:研究顺铂在体外诱导宫颈癌Hela细胞凋亡及其作用机制。方法:采用MTT法测定顺铂对Hela细胞增殖的影响;流式细胞仪和Hochest33258检测药物作用前后的细胞凋亡情况;RT-PCR检测HPVE6的mRNA水平表达;WesternBlot测定HPVE6、p53、p21、Bax、Bcl-2蛋白水平的表达。结果:顺铂抑制Hela细胞生长呈时效和量效关系;经10μg/ml顺铂分别在12、24、36、48小时作用Hela细胞,亚G1峰与对照组有显著性差异;RT-PCR提示顺铂作用Hela细胞后HPVE6的mRNA水平表达逐渐降低;WesternBlot提示顺铂作用Hela细胞后HPVE6的蛋白水平表达逐渐降低,p53、p21和Bax蛋白水平表达逐步升高,Bcl-2的表达无变化。结论:顺铂通过抑制HPVE6的表达,恢复p53的功能,引起细胞凋亡,起到杀伤肿瘤的作用。     

丹皮酚协同顺铂抗人宫颈鳞癌SiHa细胞及其机制

[目的]探讨丹皮酚（paeonol,Pae）和顺铂（CDDP）联合用药对人宫颈癌SiHa细胞的增殖抑制、凋亡诱导作用。[方法]应用MTT法检测不同浓度的Pae或CDDP单独及联合用药对宫颈癌SiHa细胞增殖的抑制作用,同时观察Pae与CDDP的协同抗肿瘤作用。流式细胞仪测定Pae联合CDDP用药对SiHa细胞凋亡的诱导作用。[结果]各浓度Pae（9.375～300mg/L）和CDDP（0.625～10mg/L）对人宫颈癌SiHa细胞均有明显增殖抑制作用,且呈时间—剂量依赖效应关系。18.75、37.5、75mg/LPae分别与2.5、5、75mg/LCDDP联用时具有协同作用,且以75mg/LPae与5mg/LCDDP联用时协同作用最显著（CDI=0.544）。75mg/LPae组细胞凋亡率为8.26%±1.12%,5mg/LCDDP组细胞凋亡率为29.62%±2.48%,与75mg/LPae联合5mg/LCDDP组细胞凋亡率（52.49%±4.39%）比较均有显著性差异（P〈0.01）。[结论]Pae与CDDP联合应用具有显著的协同抗人宫颈癌SiHa细胞增殖作用,其机制可能与两者协同诱导SiHa细胞凋亡有关。     

咖啡因与苯巴比妥联合对顺铂的体外抗癌增效作用

目的：研究咖啡因（CAF）对顺铂（DDP）的抗癌增效作用及苯巴比妥（PBT）是否抑制咖啡因的抗癌增效作用。方法：以SPC－A－1细胞为,要用MTT法研究甲基黄嘌呤类药物－CAF及其与PBT联合应用对DDP的体外细胞毒作用的影响,结果：120ug/ml的CAF及10ug/ml PBT对细胞生长无明显的细胞毒作用,120ug/ml的CAF可明显增强DDP对SPC－A－1细胞的细胞毒作用（P＜0．05）,10ug/ml PBT对DDP无增效作用（P＞0．05）,120ug/ml CAF与10ug/ml PBT的混合物也能增强DDP的细胞毒作用（P＜0．01）,其增效作用与单纯CAF的增效作用比较无显著性差异（P＞0．05）,。结论：咖啡因可明显增强DDP对SPC－A－1细胞的体外抗癌效应,而苯巴比妥对咖啡因的体外抗癌增效作用无拮抗作用,提示苯巴比妥作为降低咖啡因副反应的成分,有可能与咖啡因联合作用作为化疗增效剂用于肺癌治疗。     

顺铂对肝癌细胞凋亡及其细胞周期的影响

目的：探讨顺铂对Hca-F细胞凋亡及细胞周期的影响。方法：采用荧光，透射电镜及流式细胞术分析法检测肝癌细胞凋亡，细胞周期和增殖指数（PI）的变化。结果：CDDP可引起肿瘤细胞凋亡，并随着给药时间的延长，药物浓度的增高，细胞凋亡率随之增高，细胞凋亡形态结构越发典型，表现为核固缩，染色质边集，核小体形成等，同时药物浓度的升高，细胞周期S期比例下降，G0/G1期比例上升，PI增殖指数下降。结论：CDDP可引起细胞凋亡，细胞周期G0/G1期阻滞，使细胞分裂增殖活动受到抑制。     

奈达铂体外诱导人卵巢癌顺铂耐药细胞凋亡的机制

目的探讨研究奈达铂（NDP）体外对人卵巢癌顺铂原发耐药细胞株SKOV3和继发耐药株SKOV3/DDP的抑制作用及其机制。方法采用MTT法检测体外不同时间不同浓度的NDP对SKOV3及SKOV3/DDP细胞的杀伤作用,计算半数抑制浓度(IC50);流式细胞术(FCM)测定给药前后细胞凋亡率及周期分布变化;半定量PCR分析凋亡基因Bcl-2,Bax,caspase-3,caspase-9及反应肿瘤细胞侵袭力的基因MMP-2的表达变化。结果NDP能明显抑制SKOV3及SKOV3/DDP生长,呈时间-剂量依赖性;流式结果显示随时间、浓度增加,SKOV3和SKOV3/DDP细胞凋亡明显增加,S期细胞的比例增加,差异均有统计学意义（P<0.05）。与对照组相比,Bcl-2、MMP-2表达减少,Bax,caspase-3,caspase-9表达增加。结论NDP可抑制SKOV3和SKOV3/DDP细胞增殖,其机制可能与诱导细胞凋亡,细胞S期阻滞,下调Bcl-2及上调Bax,caspase-3,caspase-9的表达有关;同时可下调MMP-2的表达,有利于降低卵巢癌细胞的侵袭力。     

Comparative Studies to Evaluate Relative in vitro Potency ofLuteolin in Inducing Cell Cycle Arrest and Apoptosis in HaCaTand A375 Cells

Luteolin is a naturally occurring flavonoid present in many plants with diverse applications in pharmacology.Despite several studies elucidating its significant anti-cancer activity against various cancer cells, the mechanismof action in skin cancer is not well addressed. Hence, we investigated the effects of luteolin in HaCaT (humanimmortalized keratinocytes) and A375 (human melanoma) cells. The radical scavenging abilities of luteolin weredetermined spectrophotometrically, prior to a cytotoxic study (XTT assay). Inhibitory effects were assessedby colony formation assay. Further, the capability of luteolin to induce cell cycle arrest and apoptosis weredemonstrated by flow cytometry and cellular DNA fragmentation ELISA, respectively. The results revealedthat luteolin possesses considerable cytotoxicity against both HaCaT and A375 cells with IC50 values of 37.1μM and 115.1 μM, respectively. Luteolin also inhibited colony formation and induced apoptosis in a dose andtime-dependent manner by disturbing cellular integrity as evident from morphological evaluation by Wright-Giemsa staining. Accumulation of cells in G2/M (0.83-8.14%) phase for HaCaT cells and G0/G1 (60.4-72.6%)phase for A375 cells after 24 h treatment indicated cell cycle arresting potential of this flavonoid. These datasuggest that luteolin inhibits cell proliferation and promotes cell cycle arrest and apoptosis in skin cancer cellswith possible involvement of programmed cell death, providing a substantial basis for it to be developed into apotent chemopreventive template for skin cancer.     

热疗合用顺铂的体外热动力学研究

用ＭＴＴ比色法进行热疗联合顺铂对人胃癌细胞ＭＫＮ２８细胞毒作用的动力学研究发现，随着温度的升高和加温时间的延长，细胞毒作用增强；４３℃３０分钟以上的热疗有较强的细胞毒作用；４３℃３０分钟以下的热疗作用较弱，与顺铂合用有明显的协同作用，二者同时合用的效果优于热疗后化疗或先化疗后热疗。     

Genotoxicity and Interference with Cell Cycle Activities by an Ethanolic Extract from Thai Plumbago indica Roots in Human Lymphocytes in vitro

In Thai traditional medicine, Plumbago indica or Jetamul-Pleung-Dang in Thai is known to have healthbenefit especially for anti-inflammatory, antibacterial, and antitumor activities. However, the mechanisms ofits action are still uncertain. One of which might be genotoxic effects. In the present study, we investigated thegenotoxicity of an ethanolic extract of Plumbago indica root (EEPIR) by sister chromatid exchange (SCE) assayin human lymphocytes. Results have shown that all treatments with EEPIR (12.5-100 μg/ml) could induce cellcycle delay as shown by significant increase in the number of metaphase cells in the first cell cycle but neither inthe second nor the third cell cycle. Only at concentrations of 25, 50, and 100 μg/ml were SCE levels significantlyincreased above that of the control (p<0.05) . EEPIR at a concentration of 500 μg/ml induced cell death as fewmitotic cells were shown. Accordingly, EEPIR (25-100 μg/ml) is genotoxic in human lymphocytes and cytotoxic atconcentrations of ≥500 μg/ml in vitro. Therefore, these activities of the EEPIR could serve its potential therapeuticeffects, especially as an anticancer agent. Further study of EEPIR in vivo is now needed to support this in vitroevidence.     

家蝇抗菌肽Cecropin对人肝癌BEL-7402细胞周期的影响及其机制

目的探讨家蝇抗菌肽Cecropin对人肝癌BEL-7402细胞周期的影响及其机制。方法人肝癌细胞BEL-7402经家蝇抗菌肽Cecropin作用后,光学显微镜观察细胞生长情况,PI单染流式细胞术检测各组细胞周期分布情况,并采用间接免疫荧光标记技术通过流式细胞仪检测细胞周期检测点激酶1(CHK1) 和细胞周期依赖性蛋白激酶2(CDK2)。结果光学显微镜下,对照组细胞形态规则,呈梭形贴壁生长,抗菌肽组细胞部分变圆,贴壁不佳,数量减少;细胞增殖周期发生改变, G₀/G₁与G₂/M期细胞比例降低,S期细胞比例增高,与对照组相比差异有统计学意义(P<0.05) ,并且细胞周期调控关键蛋白分子CHK1和CDK2的表达增高。结论家蝇抗菌肽Cecropin可通过调节CHK1和CDK2的表达影响BEL-7402细胞增殖周期。     

半边旗抗肿瘤有效成分6F对HL-60细胞周期的影响及体外增效作用

目的：研究半边旗抗肿瘤有效成分６Ｆ对ＨＬ－６０细胞周期的影响及对常用抗肿瘤药的体外增效作用。方法：应用流式细胞光度术（ＦＣＭ）测定细胞周期，应用噻唑蓝（ＭＴＴ）法测定药物对细胞的抑制率。结果：不同浓度６Ｆ作用６小时即可使ＨＬ－６０细胞Ｓ期及Ｇ２／Ｍ期比例升高，Ｇ１期比例下降，并呈一定的剂量效应关系，当作用到２４小时后，Ｓ期比例进一步升高，但Ｇ２／Ｍ期比例稍有回落。低浓度６Ｆ分别与２－氯代脱氧腺苷（２－ＣＬｄＡｄｏ），顺铂（ＣＤＤＰ），长春新碱（ＶＣＲ），氟尿嘧啶（５ＦＵ）合用可增强它们对ＨＬ－６０细胞的杀伤作用，ｑ值大于０．８５，与各药有相加或协同作用，６Ｆ对２－ＣｌｄＡｄｏ，ＣＤＤＰ，ＶＣＲ，５ＦＵ的增效倍数分别为１．５８，１．５３，１．５５，１．３８。结论：６Ｆ可明显阻断ＨＬ－６０细胞在Ｓ期及Ｇ２／Ｍ期；６Ｆ可增强上述药物对ＨＬ－６０细胞的杀伤作用。已知，２－ＣｌｄＡｄｏ阻断细胞在Ｓ期，ＣＤＤＰ和ＶＣＲ阻断Ｇ２／Ｍ期，５ＦＵ阻断Ｇ１期。鉴于所试药物对细胞周期的影响不同，提示６Ｆ的体外增效作用可能与此有关。     

Temperature-sensitive Ovarian Carcinoma Cell Line (OvBH-1)

OvBH-1 cells from a patient with ovarian clear cell carcinoma were established and their biochemical status was analyzed. Cells grown at 37°C exhibited normal cell cycle distribution, whereas the cells shifted to 31°C were arrested in the G₂/M phase of the cell cycle. Immunochemical analysis using anti-p53 antibodies (DO-1, PAb240, PAb421, and PAbl620) revealed that only the DO-1 antibody reacted with p53 with a high and similar percentage at both temperatures. PAb240 reacted with a low percentage of cells at 37°C and no reaction was observed at 31°C. PAb421 antibody stained a significantly lower percentage of cells at 37°C than at 31°C. Cells were not stained with PAbl620 antibody and were negative for antibodies against p21^WAF1 and MDM2 proteins independently of the temperature. Sequencing of all coding exons of the p53 gene demonstrated only a neutral genetic polymorphism, i.e. a G-to-A substitution (GAG to GAA) at nucleotide position 13 432. Thus, the observed temperature sensitivity of OvBH-1 cells cannot be ascribed to a p53 primary structure mutation. Based upon immunochemical analyses, we consider, however, that p53 in nuclei of OvBH-1 cells is in a highly unstable conformation. Furthermore, the N-terminal portion of the p53 protein at Ser20 has not been modified, and Lys373 and/or Ser378 of the C-terminus is acetylated and/or phosphorylated. The nuclear location signal of p53 is preserved. Induction of MDM2 protein is uncoupled from the cell regulatory machinery and the induction of p21^WAF1 by p53 is unpaired in OvBH-1 cells.     

新型光敏剂联合阿霉素对乳腺癌细胞增殖及周期的影响

目的:研究新型光敏剂叶绿素衍生物(Chlorophyl derivative4,CPD4),联合阿霉素对人乳腺癌MCF-7细胞增殖及周期的影响,初步探讨联合用药的作用机制,为临床开辟新的治疗方法提供实验依据.方法:以人乳腺癌MCF-7细胞系为研究对象,新型光敏剂和乳腺癌传统化疗药物阿霉素(ADM)联合给药.采用流式细胞仪检测ADM组(2个浓度组分别预处理24小时、48小时)、光动力效应组、联合用药组细胞凋亡率和周期分布:流式细胞仪分析ADM(20ng/mL)预处理24小时、48小时对细胞平均荧光强度的影响以及ADM预处理24小时、48小时后加入CPD4 1.5μg/mL孵育不同时间细胞平均荧光强度的变化.结果:联合用药组细胞凋亡率明显高于单药组,差异有统计学意义(P＜0.01);光动力效应可造成MCF-7细胞G_0/G_1期阻滞,低浓度ADM预处理后GdM期细胞增加,联合用药时G_2/M期细胞升高.ADM预处理MCF-7细胞24小时、48小时细胞平均荧光强度与对照组荧光强度比较差异无统计学意义(P＞0.05);ADM预处理MCF-7后能增加光敏剂CPD4进入细胞的量,在CPD4孵育2小时细胞平均荧光强度最强,且ADM预处理48小时组＞预处理24小时组＞对照组.结论:ADM预处理MCF-7细胞后能够增加光敏剂CPD4进入细胞的量;光动力效应联合ADM具有协同作用.     

Cell cycle and apoptotic effects of SAHA are regulated by the cellular microenvironment in HCT116 multicellular tumour spheroids

Using multicellular tumour spheroids (MCTS) of HCT116 colon carcinoma cells, we analysed the effects of SAHA (suberoylanilide hydroxamic acid), a histone deacetylase inhibitor (HDACi). We found that, although SAHA-induced histone acetylation and ROS level upregulation occur throughout the spheroid, inhibition of cell cycle progression and induction of apoptosis are dependent on cell microenvironment. SAHA-induced growth inhibition of HCT116 MCTS results from the inhibition of cell cycle progression and induction of apoptosis. At a low concentration SAHA decreases Ki-67 and cyclin A positive cells and increases p21 positive cells in the outer layer while it induces a ROS-dependent apoptosis in the central zone of the spheroid. Coimmunostaining of p21 and apoptotic cells confirms that SAHA effects are different depending on the position of the cells within the spheroid. At a higher dose, SAHA induces mitotic defects and survivin downregulation in the outer layer of cells resulting in an additional cytotoxic effect in this part of the spheroid. Together these findings show that SAHA-induced cytostatic and cytotoxic effects occur in different cell populations, indicating that the cellular microenvironment is an important determinant in the regulation of the effects of SAHA treatment. Consequently, the MCTS model appears to be a valuable advanced tool for evaluating the effects of SAHA treatment in combination with other anticancer agents.