CDK1 siRNA干扰在Taxol诱导细胞凋亡中的作用

目的 研究转染细胞周期依赖性蛋白激酶1(cyclin-dependent kinase 1,CDK1)siRNA、以及转染后进行凋亡刺激对细胞周期和凋亡的影响,探讨CDK1在细胞凋亡中的确切作用,揭示细胞周期与细胞凋亡协调的分子机制.方法 以人宫颈癌细胞株HeLa细胞为研究对象,脂质体转染CDK1 siRNA,转染后48 h加紫杉醇(Taxol) (20 μg/ml)刺激凋亡,Western印迹检测CDK1和抗凋亡蛋白BCL2表达,AnnexinV/PI法检测细胞的凋亡,流式细胞仪分析DNA含量检测细胞周期.结果 转染CDK1 siRNA后,CDK1蛋白的表达下降,细胞周期G2/M期比例增加,细胞凋亡率与对照相比没有明显升高.只加Taxol刺激12 h后细胞凋亡率增加并伴有S期和G2/M期比例增加. 转染CDK1 siRNA后再用Taxol刺激,其细胞凋亡率没有明显改变,G2/M期阻滞效应也没有叠加.BCL2蛋白只在加Taxol刺激组表达下降,与CDK1表达减少没有相关性.结论 siRNA沉默导致的CDK1表达降低只导致细胞周期G2/M期阻滞,没有引起细胞凋亡;CDK1的表达降低对紫杉醇所诱导的细胞周期阻滞和细胞凋亡效应没有明显影响.     

二烯丙基二硫化物诱导人肺癌H1299细胞凋亡机理研究

目的:研究大蒜提取物二烯丙基二硫化物(DADS)诱导人非小细胞性肺癌细胞H1299凋亡及细胞周期阻滞对此过程的影响,探讨DADS抗肿瘤的可能机制。方法:MTT法检测细胞活性、Hoechst 33258染色法计数凋亡细胞、流式细胞术检测细胞凋亡率及周期分布。结果:与未处理的对照组相比,DADS可诱导H1299细胞产生典型的凋亡细胞形态学变化,并呈浓度依赖性诱导H1299细胞凋亡;较高浓度的DADS能诱导G2/M期细胞百分率显著升高。结论:DADS能诱导H1299细胞发生细胞凋亡,并诱导H1299细胞发生G2/M期阻滞。     

厄贝沙坦通过MAPK-ERK1/2信号通路影响乳腺癌MCF-7细胞的增殖

目的:初步研究血管紧张素Ⅱ1型受体(AT1R)阻断剂对人乳腺癌细胞系MCF-7的细胞增殖的抑制作用及其机制。方法:MTT法观察血管紧张素Ⅱ1型受体阻滞剂(ARBs)厄贝沙坦对MCF-7细胞增殖的抑制作用;采用流式细胞术分析厄贝沙坦对MCF-7细胞周期及凋亡的影响;应用Westernblot技术分别检测ERK1/2及p-ERK1/2蛋白。结果:厄贝沙坦对MCF-7细胞具有明显的抑制作用,并具有剂量依赖性;厄贝沙坦影响MCF-7细胞的周期,使得细胞的G0/G1期细胞比例增多(P<0.05),而S期细胞比例下降(P<0.05),抑制细胞的生长;但与对照组相比,只有高浓度的厄贝沙坦(1×10-4mol/L)具有促进细胞凋亡的作用(P<0.05);厄贝沙坦可抑制MCF-7细胞p-ERK蛋白的表达(P<0.05),但不影响总ERK1/2蛋白的表达(P>0.05)。结论:厄贝沙坦可以通过影响ERK1/2 MAPK信号转导通路抑制MCF-7乳腺癌细胞的生长。     

红景天苷对小鼠MSCs的增殖及其细胞周期的影响

目的本研究探讨了不同浓度红景天苷通过ERK信号通路对小鼠骨髓间充质干细胞增殖和细胞周期变化的影响及机制。方法实验分为对照组(D-MEM/F-12完全培养液)、不同浓度红景天苷(5~100μg/m L)诱导组和阻断组(5~100μg/m L+30μg/L PD98059),利用CCK8法、流式细胞术及Western blot分别检测ERK信号通路阻断前后细胞抑制率、细胞周期及其相关蛋白的表达。结果 10μg/m L诱导组和阻断组均能抑制细胞增殖,且5μg/m L(P0.01)、25μg/m L(P0.05)和100μg/m L(P0.05)诱导组对细胞抑制显著高于阻断组。25μg/m L诱导组和阻断组G_0/G_1期细胞比例均增加,细胞周期阻滞于G0/G1期。诱导组和阻断组cyclin A和cyclin D1蛋白的表达与对照组比较均下调,P21蛋白的表达上调。阻断组cyclin A蛋白的表达与诱导组比较明显下调(P0.01);50和100μg/m L诱导组与阻断组相比cyclin D1蛋白表达显著下调,P21蛋白的表达显著上调(P0.01)。结论红景天苷能通过调节细胞周期相关蛋白的表达抑制小鼠MSCs细胞的增殖和细胞周期的改变。     

TLR9激活后影响人非小细胞肺癌细胞对多西他赛化疗敏感性的体外实验

目的:探讨含非甲基化Cp G基序的寡脱氧核苷酸(Cp G oligodeoxyribonucleotides 7909,Cp G ODN7909)与Toll样受体(Toll-like receptor,TLR)9结合后是否影响人肺癌A549和H520细胞对多西他赛(doctaxel,DOC)的化疗敏感性及相关机制。方法:设计并合成特异性干扰siRNA序列,转染细胞,Western blot法检测TLR9 siRNA的沉默效果,CCK-8法检测细胞活力。设空白对照组、阴性对照组和TLR9 siRNA干扰组,分别接受Cp G ODN7909和多西他赛单独或联合治疗,流式细胞术检测细胞周期及凋亡,Western blot法检测P38及Bax蛋白表达。结果:在2种细胞中,Cp G ODN7909单独处理后,细胞活力和凋亡率都无明显变化,G2/M期细胞比例P38和Bax蛋白的表达明显升高(P0.01);Cp G ODN7909处理TLR9 siRNA干扰细胞各指标均无明显变化。多西他赛单独处理后3组细胞的细胞活力明显下降,G2/M期细胞比例、凋亡率和Bax蛋白表达都明显升高(P0.01),P38表达无明显变化。与多西他赛单独处理比较,多西他赛与Cp G ODN7909联合治疗后细胞活力明显下降,G2/M期细胞比例、凋亡率和Bax蛋白表达都明显升高(P0.01),P38表达无明显变化;联合处理TLR9 siRNA干扰细胞各指标均无明显变化。结论:Cp G ODN7909与TLR9结合后可增强多西他赛抑制人肺癌细胞的细胞活力,诱导G2/M期阻滞,增强多西他赛对细胞的凋亡诱导作用进而提高其对多西他赛的化疗敏感性。     

Probucol抑制氧化低密度脂蛋白诱导的大鼠主动脉平滑肌细胞增殖

目的:研究Probucol抑制ox-LDL诱导RASMCs增殖与信号蛋白分子ERK1/2、MKP-1、HO-1和Trx-1表达之间的关系。方法:采用MTT、流式细胞术和Western blotting观察ox-LDL刺激条件下probucol对细胞周期、细胞增殖和凋亡、ERK1/2、MKP-1、HO-1和Trx-1表达的影响。结果:(1)Probucol抑制ox-LDL刺激RASMCs增殖:100μmol/Lprobucol+35mg/Lox-LDL组与35mg/Lox-LDL组比较,A值下降了34.9%(P0.01);(2)Probucol通过使RASMCs停滞在G0/G1期和诱导细胞凋亡2种方式抑制ox-LDL刺激细胞增殖。(3)ox-LDL显著抑制MKP-1的蛋白表达,与对照组比较下降了60.0%(P0.01),同时使p-ERK1/2表达增加了34.7%;Probucol使MKP-1蛋白表达显著增加2倍,p-ERK1/2表达降低了15.7%(P0.01);(4)35mg/Lox-LDL使细胞内Trx-1蛋白表达下降28.9%(P0.05),HO-1蛋白表达轻度增加(P0.05)。与ox-LDL组比较,probucol使Trx-1蛋白表达增加了91.6%(P0.01),HO-1表达增加31.9%(P0.01)。结论:Probucol通过增强MKP-1和HO-1蛋白表达、抑制细胞周期运转和诱导细胞凋亡的机制抑制RASMCs增殖。     

JNK抑制剂对D-氨基葡萄糖衍生物诱导人食管癌Eca-109细胞周期阻滞和凋亡的影响

目的 观察特异性C-JUN氨基末端激酶(JNK)抑制剂SP600125对D-氨基葡萄糖衍生物2-(3-羧基-1-丙酰氨基)-2-脱氧-D-葡萄糖(COPADG)诱导的Eca-109细胞凋亡和细胞周期阻滞的影响,并探讨COPADG诱导Eca-109细胞凋亡的潜在分子机制.方法 体外培养Eca-109细胞,以COPADG及SP600125与细胞作用;Western blot法检测P-JNK蛋白表达,MTT法检测细胞增殖,流式细胞术检测细胞周期.结果 COPADG显著增加Eca-109细胞P-JNK蛋白的表达和细胞凋亡率,且诱导Eca-109细胞发生G0/G1期细胞阻滞,SP600125明显减少Eca-109细胞凋亡,并使G0/G1期细胞阻滞向G2/M期细胞阻滞发展.结论 COPADG可能通过激活JNK信号通路诱导Eca-109细胞凋亡.     

棕榈酸抑制胰岛MIN6 β细胞生长的分子机制

目的:观察饱和脂肪酸棕榈酸(Palmitate,PA)对小鼠MIN6β细胞生长的影响,从细胞周期角度来探讨其发生的分子机制.方法:采用5 g/L BSA代替血清培养36 h使MIN6细胞同步化处于G0期.然后用PA(0.25～1.0 mmol/L,45 min～24 h)干预,采用MTT法检测细胞活力,流式细胞术测定细胞周期,采用Westen blot检测细胞周期相关蛋白CDK4、 CyclinD1表达水平的变化.结果:和对照组相比(1)不同浓度PA均显著抑制MIN6细胞增殖(P＜0.01);(2)PA亦能明显抑制细胞周期进程,使MIN6细胞周期更多滞留在G0/G1期(P<0.01),G2/M期与S期细胞比例降低(P<0.01);(3)PA能显著的抑制细胞周期相关蛋白CDK-4、 CyclinD1的表达(P＜0.05),并与细胞周期延迟一致.结论:PA抑制MIN6β细胞生长可能是通过降低MIN6β细胞的细胞周期相关蛋白cyclin D1/CDK-4的表达,导致从G1-S期的阻滞,从而减弱细胞增殖.     

地高辛对人胃癌SGC7901细胞生长和凋亡的影响

目的:探究地高辛对人胃癌SGC7901细胞生长和凋亡的影响,并探讨其分子机制。方法:选取人胃癌细胞SGC7901作为研究对象,采用CCK-8法检测地高辛的细胞毒作用并测定IC_(50),采用流式细胞术检测细胞凋亡和细胞周期,采用Western blot法检测c-Src、p-c-Src(Tyr416)、Akt、p-Akt(Ser473)、ERK1/2和p-ERK1/2(Tyr204)的蛋白水平,采用RT-PCR法检测c-Src的mRNA表达水平。结果:随着地高辛浓度逐渐升高,自50nmol/L开始,SGC7901细胞的存活率逐渐降低(P0.05),当地高辛浓度达到500 nmol/L时SGC7901细胞的存活率降至最低(P0.05),地高辛作用24 h的IC_(50)为191.45 nmol/L;200 nmol/L地高辛作用SGC7901细胞24 h后,与对照组相比,细胞凋亡率显著升高(P0.05),G_0/G_1期细胞比例明显升高(P0.05),c-Src、ERK1/2和Akt蛋白的磷酸化水平显著下降(P0.05),c-Src的mRNA和蛋白表达水平均明显降低(P0.05),细胞迁移率降低(P0.05)。结论:地高辛可在体外诱导人胃癌细胞SGC7901的凋亡和G0/G1期阻滞,这可能与其下调c-Src基因表达以及抑制ERK1/2和Akt蛋白磷酸化有关。     

125I粒子对食管癌Eca-109细胞凋亡及细胞周期的影响

研究,125I粒子诱导体外培养的人食管癌Eca-109细胞凋亡及对其细胞周期的影响.设置A组:对照(不加粒子),B组:低剂量(7.4×106Bq 125I粒子1枚),C组:中剂量(14.8×106Bq 125I粒子1枚),D组:高剂量(29.6×106Bq 125I粒子1枚),细胞培养1周后收集细胞,流式细胞术检测细胞凋亡及细胞周期.4组凋亡细胞阳性指数(AI)分别为0.17%、1.17%、4.35%、4.72%,B、C和D组与A组之间均有非常显著性差异(P<0.01);C组与B组,D组与B组比较,均有非常显著性差异(P<0.01),D组与C组比较无显著性差异(P>0.05).随粒子剂量增加,G2/M期细胞所占比例增高,各组之间有非常显著性差异(P<0.01).125I粒子可以诱导体外培养的人食管癌Eca-109细胞凋亡,其诱导凋亡的能力可能是通过把细胞阻滞在G2/M期实现.     

黄芩素和U0126 体外协同抗人膀胱癌细胞的作用及机制研究

目的：探讨黄芩素和U0126体外协同抗人膀胱癌T-24细胞的作用及机制研究。方法：用黄芩素、U0126及二者联合处理T-24细胞,流式细胞术检测细胞周期、细胞凋亡;显微镜下计数细胞数量变化;TUNEL法检测细胞凋亡指数;实时定量PCR和Western blot分别检测T-24 细胞胞外信号调节激酶1/2(ERK1/2) 、CyclinD1、GSK-3β和AKT ＲNA水平、蛋白水平,分析黄芩素与U0126对膀胱癌细胞凋亡和增殖的影响。结果：T-24细胞经不同浓度黄芩素处理后, 细胞凋亡率显著增加;T-24细胞经黄芩素处理24 h,G0/G1 期细胞比例明显增多,S 期细胞比例明显下降,细胞数减少,采用黄芩素联合U0126 处理T-24细胞24 h,S 期细胞比例显著降低;黄芩素或U0126单独处理T-24细胞引起明显细胞凋亡,二者联合处理凋亡更明显;利用两种药物单独或联合作用T-24细胞24 h,GSK-3β、ERK1/2、AKT磷酸化水平显著降低, ERK1/2、CyclinD1的 mRNA表达减少,联合处理作用更显著。结论：黄芩素和U0126 均可诱导T-24细胞凋亡,增加G0/G1 期细胞比例,降低S 期细胞比例,联合应用效果更明显。     

Study of the thymocyte cell cycle by bivariate analysis of incorporated bromodeoxyuridine and DNA content

The in vivo cell cycle of normal murine thymocytes was studied by bivariate analysis of bromodeoxyuridine and total DNA content in the 24 h following a single injection of the thymidine analogue. Bromodeoxyuridine incorporation was strictly limited to cells in S phase and 98% of S phase cells were labeled, demonstrating high efficiency and specificity. Cell-cycle parameters were determined by measuring the DNA content of bromodeoxyuridine-labeled cells, related to their distribution in the different phases. The changes of this distribution as a function of time reflected the progression of the cells along the cell cycle. The duration of total cycle, S phase, and both G2/M and G1 was 10 h, 6.5 h and 1.5 h, respectively. All thymocytes labeled in S phase entered G2/M, divided and returned to the G0/G1. Seventy percent of them remained in the resting state, and the other 30% re-entered a second S phase. Cell-cycle parameters of isolated CD4-CD8- cells were also determined. No evidence of cell loss during S or G2/M phase was found, suggesting that intrathymic cell death is not directly linked to the proliferative phases of differentiation.     

Effect of macrophage-derived growth factor on the cell cycle kinetics of cultured arterial smooth muscle cells

Effects of macrophage-derived growth factor on the progression of the cell cycle of cultured aortic smooth muscle cells were investigated by using rabbit peritoneal macrophage conditioned-medium (M phi-CM) as the source of the growth factors. DNA content of individual SMC in the cell population which had been exposed either to platelet-poor plasma serum (PPPS) or to M phi-CM was also measured by flow cytometry and microspectrophotometry, as well as by phase contrast microscopy. The results showed that the cells quiescent in PPPS medium had a high population not only in G0/G1 but also in G2 phase. This indicated that the cell cycle process was blocked both in G0/G1 and G2 phase. The cell cycle distribution of SMCs exposed to M phi-CM for 48 hours was very similar to that cultured in 10% fetal calf serum medium. It is suggested that M phi-CM can promote the cell growth-arrested in ppps medium passing through G0/G1 into S phase as well as through G2 into M phase.     

PD98059抑制aFGF诱导的人膀胱癌细胞株EJ细胞增殖

目的探讨细胞外信号调节激酶(ERK)激酶MEK抑制剂PD98059对酸性成纤维细胞生长因子(aFGF)诱导的人膀胱癌细胞株EJ细胞增殖的抑制作用。方法以不同浓度的aFGF和PD98059作用EJ细胞,通过细胞计数法、MTT比色法观察PD98059对细胞增殖的抑制作用;流式细胞仪测定细胞各周期细胞百分数的变化;利用[γ3-2P]ATP掺入外源性底物的方法,液体闪烁测定ERK活性的变化。结果aFGF使EJ细胞株增殖比明显增加,而PD98059使EJ细胞株增殖比明显下降,在一定浓度范围内,其程度均随浓度增高而增强。EJ细胞受到aFGF刺激后,由G0+G1期进入S和G2+M期的细胞明显增多,在PD98059的作用下,由G0+G1期进入S和G2+M期的细胞明显减少,并在一定浓度范围内成剂量依赖性,与对照组比较差异显著(P<0.05);随着aFGF浓度的增加,EJ细胞ERK活性增高,PD98059抑制细胞内ERK活性,与PD98059浓度呈剂量依赖效应。结论PD98059对aFGF引起的EJ细胞增殖具有抑制作用,aFGF可能通过激活Ras-Raf-ERK信号转导途径来调控EJ细胞增殖,PD98059可有效阻滞此传导途径。     

Topoisomerase II-alpha expression in different cell cycle phases in fresh human breast carcinomas.

Topoisomerase II-alpha (topo II alpha) is the key target enzyme for the topoisomerase inhibitor class of anti-cancer drugs. In normal cells, topo II alpha is expressed predominantly in the S/G2/M phase of the cell cycle. In malignant cells, in vitro studies have indicated that the expression of topo II alpha is both higher and less dependent on proliferation state in the cell. We studied fresh specimens from 50 cases of primary breast cancer. The expression of topo II alpha in different cell cycle phases was analyzed with two-parameter flow cytometry using the monoclonal antibody SWT3D1 and propidium iodide staining. The expression of topo II alpha was significantly higher in the S/G2/M phase of the cell cycle than in the G0/G1 phase in both DNA diploid and DNA non-diploid tumors. In 18 of 21 diploid tumors, and in 25 of 29 non-diploid tumors, >50% of the topo II alpha-positive cells were in the G0/G1 phase. This significant expression of topo II alpha in the G0/G1 phase of the cell cycle may have clinically important implications for treatment efficacy of topoisomerase II inhibitors.     

The action of epidermal growth factor (EGF) is limited to specific phases of the cell cycle in an EGF dependent colonic cell line

In the presence of epidermal growth factor (EGF) a human colon cell line, LIM 1215, proliferates in serum-free medium. Under these culture conditions the cells are dependent on the presence of EGF for both proliferation and survival. In order to study the action of growth factors at different stages of the LIM 1215 cell cycle, pure populations of G1, S and G2/M cells were obtained by cell sorting after supravital staining of the DNA with Hoechst 33342. Conditions were established for Hoechst 33342 staining which produced satisfactory DNA histograms and greater than 80% survival of cells. The kinetics of passage for sorted S or G2/M cells into G1 were not affected by EGF or fetal calf serum. After sorting there appeared to be a 4 h delay before the cells proceeded in the cell cycle. Sorted S cells entered G2 over an 8 h period and maintained this same transition period from G2 into G1. If EGF or serum was present, these cells then re-entered the cell cycle after a variable delay and in an asynchronous manner. EGF was applied to S phase and G2/M phase LIM 1215 cells for periods of 2-10 h at various times after replating in serum-free conditions. Cells in S phase only responded to EGF as they passed from G2/M into G1. Exposure to EGF in S phase resulted in little growth stimulus once the cells returned to G1. For cells in G2/M phase, EGF was required immediately to give the maximum stimulus for re-entering the cell cycle. If the EGF was delayed for more than 8 h, the cells did not re-enter the cycle within the following 20 h. Exposure to EGF for less than 2 h failed to stimulate proliferation. These results indicate that EGF must be present as cells enter G1 from mitosis. Once the cells have entered G1, EGF is required for a 10 h period for a large number of cells to re-enter the cycle from G1.     

肠道病毒EV71相关受体在人二倍体细胞KMB17表面的构成分析

目的研究人二倍体细胞KMB17表面与肠道病毒EV71感染相关的受体Toll样受体4（TLR4）、血小板衍生生长因子受体α多肽（pDGFR—αR1）的表达情况，以及细胞周期、细胞代次、EV71感染对这两种受体表达的影响。方法使用荧光标记的特异抗体：PE—TLR4、FITC—PDGFR—αR1标记不同细胞代次、处于不同细胞周期时段的KMB27细胞，应用流式细胞仪检测表达这两种受体蛋白的阳性细胞率，以及EV71感染前后，阳性细胞率的变化。使用NCB Iprimer blast设计TLR4、PDGFR—αR1特异的引物，提取不同细胞代次、细胞周期时段KMB17细胞的RNA，使用real-time PCR检测这些受体蛋白特异mRNA的量。结果KMB17细胞在使用血清饥饿法进行G0／G1同步化后约22h进入S期，在36h进入G2期，在48h左右完成分裂。PDGFR—αR1、TLR4平均阳性细胞的比例分别为4．47％和11．82％：通过流式细胞仪检测，在G0／G1期，PDGFR—αR1、TLR4的平均阳性细胞比例分别为4％和8．9％；在S期，分别为2．6％和9％；在G2／M期，分别为4-4％和13．5％；EV71病毒感染后，PDGFR—αR1阳性的细胞比率由2．6％上升至4．0％，TLR4阳性细胞比率分别为11．6％和12．8％。结论人二倍体细胞KMB17表面表达肠道病毒相关的受体TLR4和PDGFR-αR1，表达PDGFR—αR1的阳性细胞较TLR4少，不同代次的KMB17细胞这2种受体阳性细胞比例的差异不大，PDGFR—αR1阳性细胞在GO期较多，TLR4阳性细胞在G2／M期较多；EV71病毒感染后，PDGFR—αR1、TLR4阳性细胞的比率变化不大，表明这两种受体分子在KMB17细胞表面表达较稳定，不受EV71感染的影响。     

Different accumulation of activated extracellular signal-regulated kinases (ERK 1/2) and role in cell-cycle alterations by epidermal growth factor, hydrogen peroxide, or asbestos in pulmonary epithelial cells

The extracellular signal-regulated kinase (ERK) pathway is induced by cytokines and oxidative stress. In this study we examined the patterns of localization of phosphorylated ERK proteins in relationship to subsequent phenotypic responses by the mitogenic agent epidermal growth factor (EGF) (5 ng/ ml); hydrogen peroxide (H(2)O(2)) (100 to 300 microM), an inducer of apoptosis; and crocidolite asbestos (5 microg/cm(2) dish) in a nontransformed murine alveolar type II epithelial cell line (C10). Laser scanning cytometry and flow cytometry were used to determine: (1) whether expression of phosphorylated ERKs was cell cycle-related; and (2) whether cell-cycle alterations by agents could be modified after addition of the mitogen-activated protein kinase/ERK kinase (MEK) 1 inhibitor PD98059. In contrast to other stimuli which induced transient increases in phosphorylated ERKs, asbestos caused fiber-associated localization of phosphorylated ERKs that were elevated from 1 to 24 h (P < or = 0.05), and striking apoptosis followed by increased numbers of cells in the S phase at 72 h. In both control and asbestos-exposed cells, the percentage of phosphorylated ERK-positive cells was greatest in cells in the G(2)/M and S phases of the cell cycle. All stimuli caused increased proportions of cells in G(2)/M at 24 h that were inhibited by PD98059 (30 microM). Increases in G(2)/M cells by H(2)O(2) and asbestos also were decreased at 48 h by the MEK1 inhibitor. In addition, PD98059 abrogated elevations in S-phase cells by EGF and H(2)O(2) at 24 h and by asbestos at 72 h. Our results suggest that ERKs mediate cell-cycle alterations during the development of epithelial cell apoptosis or proliferation by diverse ERK stimuli.     

陡脉冲对恶性肿瘤细胞增殖及细胞周期的影响

观察了陡脉冲对恶性肿瘤细胞生长抑制作用及其细胞周期的影响。人卵巢腺癌 SKOV3细胞受不同剂量的陡脉冲作用后 ,分别制作细胞生长曲线和应用流式细胞术检测肿瘤细胞增殖及其对细胞周期的影响。经陡脉冲作用后的癌细胞的生长受到不同程度的抑制 ,随着陡脉冲剂量的增加 ,癌细胞的死亡率和抑制效应更加显著 ;陡脉冲选择性作用于肿瘤细胞 DNA合成、分裂期 ,出现 G0 / G1 期阻滞 ,抑制肿瘤细胞的增殖 ,陡脉冲剂量较大的实验组和对照组间比较有显著性 (P<0 .0 1)。陡脉冲对恶性肿瘤细胞具有杀伤和抑制作用 ,为其用于肿瘤治疗提供依据。     

细胞外信号调节激酶1/2的阻断剂PD98059对小鼠淋巴细胞增殖的影响

目的探讨细胞外信号调节激酶1/2(ERK1/2)信号通路的特异性阻断剂PD98059对小鼠淋巴细胞体外增殖的影响。方法分离小鼠淋巴结细胞,藉羧基荧光素乙酰乙酸(CFDA-SE)染色,用多克隆刺激剂刀豆A蛋白(ConA)或者佛波醇酯(PDB)加离子霉素(Ion)刺激,以流式细胞术分析PD98059对淋巴细胞增殖的影响;利用碘化丙锭(PI)染色分析PD98059对细胞周期的影响。结果CFDA-SE染色分析显示,当浓度为5、10、20、30、40μmol/L时,PD98059对ConA诱导的淋巴细胞增殖具有明显的抑制作用,并呈现明显的剂量依赖关系(r=0.985,P<0.01)。选择最佳浓度30μmol/L的PD98059,观察其对不同时间淋巴细胞增殖的影响,结果发现,随着时间的延长,PD98059对淋巴细胞增殖的抑制作用明显降低(P<0.01),但抑制率随时间的延长而明显升高。细胞周期分析进一步表明,PD98059可阻止ConA刺激的淋巴细胞进入S期和G2/M期,而亚二倍体峰变化并不明显。PD98059对PDB Ion刺激的淋巴细胞细胞周期的影响与ConA刺激相似,不同的是S期和G2/M期的变化较ConA的作用更明显。结论PD98059对小鼠淋巴细胞的增殖有明显地抑制作用,并可阻止其进入S期和G2/M期,提示ERK1/2信号通路的活化在淋巴细胞增殖中起重要作用。