枸杞多糖对人前列腺癌PC-3细胞凋亡的影响

目的研究枸杞多糖(LBP)对人前列腺癌PC-3细胞凋亡的影响。方法体外培养的人前列腺癌PC-3细胞经枸(LBP)处理后,采用MTT法测定细胞的增殖情况,流式细胞仪分析LBP对PC-3细胞周期和凋亡的影响,TUNEL法观察PC-3细胞凋亡的变化,用免疫组化法检测其Bcl-2和Bax蛋白的表达。结果LBP可明显抑制人前列腺癌PC-3细胞的生长,并能诱导PC-3细胞凋亡,凋亡率分别为8.5%、17.5%、29.5%、37.5%和41.5%,与对照组比较,差异有非常显著性(P<0.01);同时PC-3细胞Bcl-2/Bax蛋白比值显著下降,呈明显的剂量-效应关系。结论LBP对人前列腺癌PC-3细胞DNA具有损伤作用,能诱导PC-3细胞的凋亡,调节其相关基因的表达。     

丁香酚诱导癌细胞PC-12凋亡的研究

目的 研究丁香酚诱导大鼠嗜铬细胞瘤PC-12的凋亡作用机制。方法 用不同浓度的丁香酚处理PC-12细胞,MTT法检测细胞相对存活率,荧光染色观察细胞凋亡情况;比色法检测Caspase-3酶活性;real-time PCR检测Bax和Bcl-2 mRNA水平的变化,Western blot检测蛋白水平的变化。结果 丁香酚能显著降低PC-12细胞存活率,促进Bax mRNA及蛋白表达,抑制Bcl-2 mRNA及蛋白表达,进而提高Caspase-3酶活性。结论 丁香酚可能通过上调Bax、下调Bcl-2表达,激活Caspase-3酶活性,诱导PC-12细胞凋亡。     

姜黄素对前列腺癌PC-3M细胞生长抑制和凋亡的影响

为探讨姜黄素对人前列腺癌PC-3M细胞生长抑制及凋亡调控的作用,按姜黄素浓度分成10、20、30、40μmol/L4个处理组和对照组(未加姜黄素),处理PC-3M细胞不同时间后,倒置显微镜观察细胞形态;采用MTT法检测细胞的增殖抑制情况;荧光染色法检测细胞凋亡;流式细胞仪进行细胞周期时相分析;免疫组化SP法检测细胞内caspase-3蛋白的表达水平。结果表明,与对照组比较,姜黄素处理组可使细胞明显变圆,体积缩小,脱壁细胞增多;MTT法检测显示,姜黄素处理组随着浓度增加和作用时间延长,对PC-3M细胞的增殖抑制作用增强,并呈时间、剂量依赖性(P<0.01);荧光显微镜下部分细胞发生凋亡形态学改变,对照组和4个处理组凋亡率分别为(9.83±1.53)%、(19.50±1.00)%、(24.83±2.52)%、(30.17±2.08)%和(38.50±2.65)%;流式细胞仪显示,处理组停滞在S、G2/M期细胞增加,而G0/G1期细胞减少;免疫组化结果显示,随着姜黄素浓度增加,caspase-3蛋白表达增强,呈剂量依赖性(P<0.01)。结论:姜黄素对人前列腺癌PC-3M细胞的生长具有明显抑制作用,上调caspase-3蛋白的表达,诱导细胞凋亡可能是其作用机制之一。     

葡萄汁对人前列腺癌PC-3细胞的凋亡作用

目的研究葡萄汁对人前列腺癌PC-3细胞的影响。方法将低、中、高剂量(分别为16、32和64μl/ml)的葡萄汁加入体外培养的人前列腺癌PC-3细胞,检测葡萄汁对PC-3细胞的毒性作用,用单细胞凝胶电泳测其对DNA损伤作用,流式细胞术测细胞凋亡,免疫组化法测凋亡相关基因Bax、Bcl-2的表达。结果3种剂量的葡萄汁能抑制人前列腺癌PC-3细胞增殖;DNA迁移长度与彗星细胞拖尾率逐渐增高,拖尾细胞率最高为71%;能诱导人前列腺癌PC-3细胞凋亡,细胞凋亡率分别是14.5%3、2.1%和40.5%,促凋亡基因Bax表达率分别是36.8%、50.4%和64.8%;抑凋亡基因Bcl-2表达率依次为39.1%、25.0%和12.4%。结论葡萄汁可抑制人前列腺癌PC-3细胞增长,诱导其细胞凋亡,能上调促凋亡基因Bax的表达,下调抑凋亡基因Bcl-2的表达。     

TFAR19蛋白协同米非司酮对前列腺癌PC-3M细胞凋亡的影响

目的初步探讨TFAR19协同米非司酮(MIF)对前列腺癌PC-3M细胞凋亡的影响。方法构建TFAR19真核表达载体,用脂质体介导的方法转染PC-3M细胞。MTT法检测5、10、20、50和100μmol·L-1MIF作用于前列腺癌PC-3M细胞24~96h的吸光度(A)值。在转染TFAR19的细胞中加入20mol·L-1MIF培养24、48h,MTT比色法检测细胞增殖,原位末端标记(TUNEL)法检测细胞凋亡率,透射电镜进一步观察细胞超微结构的改变。结果构建了PCI-neo-TFAR19真核表达载体并在转染的PC-3M细胞中得到了瞬时表达。MTT实验表明,与对照组相比,5、10μmol·L-1MIF组的A值差异无统计学意义(P>0.05),20、50和100μmol·L-1MIF组的A值差异有统计学意义(P<0.01),MIF对前列腺癌PC-3M细胞的抑制作用呈时间、剂量依赖性;转染PCI-neo-TFAR19并加入20 mol·L-1MIF后,与对照组及单独应用MIF组相比,细胞生长明显受到抑制(P<0.01),细胞凋亡率明显增加(P<0.01),透射电镜观察到典型的细胞凋亡特征(细胞体积缩小,核皱缩、碎裂,染色质呈块状边集等)。结论TFAR19蛋白能够协同米非司酮促进前列腺癌PC-3M细胞凋亡,有望成为前列腺癌的辅助治疗药物。     

TFARl9蛋白协同米非司酮对前列腺癌PC-3M细胞凋亡的影响

目的初步探讨TFARl9协同米非司酮（MIF）对前列腺癌PC-3M细胞凋亡的影响。方法构建TFARl9真核表达载体，用脂质体介导的方法转染PC-3M细胞。MTT法检测5、10、20、50和100μmol&#183;L^-1 MIF作用于前列腺癌PC-3M细胞24～96h的吸光度（A）值。在转染TFARl9的细胞中加入20mol&#183;L^-1 MIF培养24、48h，MTT比色法检测细胞增殖，原位末端标记（TUNEL）法检测细胞凋亡率，透射电镜进一步观察细胞超微结构的改变。结果构建了PCI-neo-TFARl9真核表达载体并在转染的PC-3M细胞中得到了瞬时表达。MTT实验表明，与对照组相比，5、10μmol&#183;L^-1 MIF组的A值差异无统计学意义（P〉0．05），20、50和100μmol&#183;L^-1MIF组的A值差异有统计学意义（P〈0．01），MIF对前列腺癌PC-3M细胞的抑制作用呈时间、剂量依赖性；转染PCI-neo-TFARl9并加入20mol&#183;L^-1 MIF后，与对照组及单独应用MIF组相比，细胞生长明显受到抑制（P〈0．01），细胞凋亡率明显增加（P〈0．01），透射电镜观察到典型的细胞凋亡特征（细胞体积缩小，核皱缩、碎裂，染色质呈块状边集等）。结论TFARl9蛋白能够协同米非司酮促进前列腺癌PC-3M细胞凋亡，有望成为前列腺癌的辅助治疗药物。     

氯化钴对PC12细胞的凋亡作用

目的　确定氯化钴(CoCl2 )对PC12细胞的影响。方法　构建CoCl2 诱导的PC12细胞模型,检测CoCl2 对PC12细胞的毒性作用;将Caspases的抑制基因p3 5转染PC12细胞得到可稳定表达p3 5基因的细胞株PC12 p3 5 ,检测p3 5对CoCl2 诱导的PC12细胞的作用;检测Caspases特异性多肽抑制剂Z VAD FMK对CoCl2 诱导的PC12细胞的作用。结果　分别以10 0、3 0 0、5 0 0、70 0和10 0 0 μmol LCoCl2 诱导PC12细胞2 4h或以5 0 0 μmol LCoCl2 分别诱导PC12细胞12、2 4、3 6、48和60h后,PC12细胞存活率均明显下降(P <0 0 1) ,并与CoCl2 诱导的时间和浓度呈正相关;细胞亚显微结构检测,流式细胞分析和DNA片段化结果也表明5 0 0 μmol LCoCl2 诱导PC12细胞2 4h可使PC12细胞出现明显凋亡特征。构建可稳定表达Caspase蛋白酶抑制基因p3 5的PC12细胞株,或分别加入5 0和10 0 μmol LCaspases多肽抑制剂Z VAD FMK预处理PC12细胞1h后,以5 0 0 μmol LCoCl2 诱导PC12细胞2 4h ,形态学观察结果,细胞存活率检测和流式细胞分析均表明p3 5基因和Z VAD FMK均可有效地抑制CoCl2 诱导的PC12细胞凋亡(P <0 0 1)。结论　CoCl2 可诱导PC12细胞凋亡。     

姜黄素对MPP~+诱导PC12细胞凋亡的影响

目的观察姜黄素对MPP+诱导的PC12细胞凋亡的影响。方法采用透射电镜,hoechst染色和流式细胞仪(FCM)观察PC12细胞凋亡,间接免疫荧光流式细胞术检测PC12细胞bcl2的表达。结果PC12细胞自然凋亡率为(15±01)%,05mmol·L-1、1mmol·L-1和2mmol·L-1MPP+作用24h后,PC12细胞凋亡率分别为(441±38)%、(549±21)%和(822±26)%;20μmol·L-1和40μmol·L-1姜黄素对PC12细胞作用24h后无明显凋亡诱导作用,但可分别使05mmol·L-1MPP+处理组细胞凋亡率由(441±38)%下降到(341±38)%和(179±15)%(P<001);PC12细胞经20μmol·L-1姜黄素处理24h后,其bcl2表达率由正常对照的(3643±790)%增加到(7673±860)%(P<001)。结论姜黄素可以抑制MPP+诱导的PC12细胞凋亡,其作用机制之一可能与促进bcl2的表达有关。     

对氨基水杨酸钠对铅诱导体外培养PC12细胞凋亡的影响

目的探讨对氨基水杨酸钠(PAS-Na)对铅诱导PC12细胞凋亡的影响。方法 PC12细胞培养至其对数生长期后,正常对照组和正常+PAS-Na 500μmol·L~(-1)组细胞于正常培养液中培养24 h后弃培养液,前者加入正常培养液、后者加入PAS-Na继续培养24 h。醋酸铅模型组及醋酸铅+PAS-Na 20,100和500μmol·L~(-1)组先与醋酸铅(终浓度10μmol·L~(-1))共培养24 h后弃培养液,再加入相应浓度PAS-Na继续培养24 h。用MTT法测定细胞存活率,试剂盒方法测定细胞内还原型谷胱甘肽(GSH)含量,Hoechst33342荧光染色检测凋亡率,AnnexinⅤ/PI双染流式细胞术检测PC12细胞早期凋亡率,Western蛋白质印迹法检测Bcl-2、Bax和P53蛋白水平。结果与正常对照组比较,醋酸铅模型组细胞存活率降低(P<0.05),细胞凋亡形态变化典型,细胞早期凋亡率增高(P<0.05),细胞内GSH含量降低(P<0.01),P53蛋白水平升高(P<0.01),Bax/Bcl-2比值升高(P<0.01);正常+PAS-Na 500μmol·L~(-1)组上述指标未见明显变化。与醋酸铅模型组比较,醋酸铅+PAS-Na组细胞存活率增高(P<0.05),细胞凋亡率和早期凋亡率均降低(P<0.05),细胞内GSH含量增高(P<0.01),P53蛋白水平和Bax/Bcl-2比值降低(P<0.05)。结论 PAS-Na对铅致PC12细胞凋亡有一定的拮抗作用,其机制可能与PAS-Na抗脂质过氧化损伤和降低Bax/Bcl-2比值有关。     

雌激素对血管平滑肌细胞凋亡的影响

目的 :观察雌激素对血管平滑肌细胞 (SMC)凋亡的影响。方法 :以大鼠为对象 ,观察正常对照组、去卵巢组及去卵巢 +雌二醇治疗组大鼠血管平滑肌细胞凋亡的关系。结果 :雌二醇治疗组血管平滑肌细胞凋亡较去卵巢组相比明显减少(P>0 .0 5 ) ,与正常对照组比较差别无显著性意义 (P >0 .0 5 )。结论 :雌二醇对血管平滑肌细胞的凋亡有抑制作用 ,可能是其预防动脉粥样硬化的机制     

Diethyl phthalate enhances expression of SIRT1 and DNMT3a during apoptosis in PC12 cells

Diethyl phthalate (DEP) works as a phthalate plasticizer and is ubiquitously used in personal care products, cosmetics, medical equipment and pharmaceutical coating. DEP is considered a potential endocrine disruptor. Previously we found DEP‐enhanced apoptosis induced by serum deprivation in PC12 cells. However, the relationship between DEP and longevity‐related factors, sirtuins and epigenetic factors (e.g. DNA methyltransferases) remains unclear, because genome modification caused by chemical toxicity, sirtuins and epigenetic factors have played key roles in abnormal metabolism and development. Here, we investigate whether DEP affects sirtuins (SIRT1 and SIRT2) and methyltranferases (DNMT1 and DNMT3a) on the apoptosis of PC12 cells. We found that DNMT3a was significantly decreased by serum deprivation. However, DNMT3a, DNMT3b and SIRT1 were significantly increased under the enhancement of apoptosis induced by serum deprivation. These results suggest that SIRT1, DNMT3a and DNMT3b play multiple and complex roles in different apoptotic stages. Our results showed DEP triggered epigenetic factors on PC12 cells apoptosis under nutrition stress. Finally, our results suggest that monitoring epigenetic factors such as DNMT3a, DNMT3b and SIRT1 could be a useful tool for chemical toxicity risk assessment. Copyright © 2012 John Wiley & Sons, Ltd.     

Zeylenone抑制人前列腺癌PC-3细胞增殖及诱导凋亡作用研究

目的研究(4R,5S,6S)-4-苯甲酰氧基-6-[(苯甲酰氧基)甲基]-5,6-二羟基-2-环己烯-1-酮Zeylenone(Zey)对人前列腺癌PC-3细胞的生长抑制和诱导凋亡作用。方法采用MTT法观察Zey对PC-3细胞和正常前列腺基质细胞WPMY-1的增殖抑制作用,应用平板克隆形成实验观察Zey对PC-3细胞克隆形成的作用,AO/EB荧光染色观察细胞形态、JC-1染色观察Zey对细胞内线粒体膜电位的影响,An-nexin V-FITC/PI双染检测凋亡细胞比率,Western blot检测凋亡相关蛋白Caspase-9、Caspase-8、Caspase-3、Caspase-7和PARP的激活及Bcl-2、Bax、Bid、Bcl-xL蛋白的表达。结果Zey可以明显抑制人前列腺癌PC-3细胞的增殖并诱导其凋亡,而对正常前列腺基质细胞WPMY-1的抑制作用显著低于PC-3细胞。Zey抑制PC-3细胞克隆形成并明显诱导其凋亡。线粒体膜电位检测结果显示Zey导致细胞内线粒体膜电位的明显降低,Western blot检测结果表明Caspase-8、Caspase-9、Caspase-7、Caspase-3被激活,PARP和Bid被剪切活化,Bcl-2、Bcl-xL表达下降,而Bax表达上调。结论 Zey可选择性抑制人非激素依赖性前列腺癌PC-3细胞的增殖并诱导其凋亡,作用机制与其对Bcl-2和Caspase家族蛋白的影响,从而诱导PC-3细胞发生内源性和外源性凋亡相关。Zey有望成为治疗前列腺癌的候选药物。     

福辛普利对平滑肌细胞凋亡及凋亡调控基因的影响

目的：探讨血管紧张素Ⅱ在不同浓度和不同作用时间下对血管平滑肌细胞凋亡及凋亡调控基因表达的作用，并研究福辛普利对其的影响。方法：采用流式细胞技术测定在不同浓度和不同作用时间血管紧张素Ⅱ作用下血管平滑肌细胞凋亡率及凋亡调控基因Fas、Bcl-2表达的变化和福辛普利对其的影响。结果：血管紧张素Ⅱ对血管平滑肌细胞凋亡和凋亡调控基因表达有较明显的诱导作用。福辛普利可抑制AngⅡ作用，对细胞凋亡有一定的影响，同时可影响凋亡调控基因的表达。结论：血管紧张素Ⅱ具有一定的的促进血管平滑肌细胞凋亡作用和调节凋亡调控基因表达的作用，尤其大剂量作用较明显。而福辛普利可明显抑制AngⅡ对血管平滑肌细胞的作用。     

石榴皮多酚对人前列腺癌PC-3细胞增殖及凋亡的影响

目的探讨石榴皮多酚对前列腺癌PC-3细胞株增殖和凋亡的影响。方法采用MTT法测定不同质量浓度组石榴皮多酚作用不同时间对PC-3细胞生长的影响,应用流式细胞仪检测石榴皮多酚作用72h后PC-3细胞的周期时相变化及凋亡情况。结果石榴皮多酚明显抑制PC-3细胞的生长,抑制效应呈时间依赖型和浓度依赖型。流式细胞仪检测结果显示,石榴皮多酚可将PC-3细胞阻滞于G1期,并诱导PC-3细胞凋亡。结论石榴皮多酚在体外对前列腺癌PC-3细胞株有明显的生长抑制和诱导凋亡的作用。     

Effects of altered food intake during pubertal development in male and female wistar rats.

The U.S. Environmental Protection Agency is currently validating assays that will be used in a Tier I Screening Battery to detect endocrine disrupting chemicals. A primary concern with the Protocols for the Assessment of Pubertal Development and Thyroid Function in Juvenile Male and Female Rats is that a nonspecific reduction in body weight (BWT) during the exposure period may potentially confound the interpretation of effects on the endocrine endpoints. Wistar rats were underfed 10, 20, 30, or 40% less than the ad libitum food consumed by controls from postnatal days (PNDs) 22 to 42 (females) or PNDs 23 to 53 (males). Terminal BWT of females and males were 2, 4, 12, and 19% and 2, 6, 9, and 19% lower than controls, respectively. In the females, neither the age of pubertal onset nor any of the thyroid hormone endpoints were affected by food restriction (FR) that led to a 12% decrease in BWT. Similarly, none of the male reproductive endpoints examined were altered by FR that led to a 9% BWT decrease. However, decreased triiodothyronine and thyroxin was observed in FR males with a 9% reduced BWT. While these data support the use of the maximum tolerated dose for BWT (10%) for the female protocol, effects on the male thyroid endpoints indicate that a slightly lower limit (相似文献   

Estrogen-like response to p-nonylphenol in human first trimester placenta and BeWo choriocarcinoma cells.

p-Nonylphenol (p-NP) is a metabolite of alkylphenol ethoxylates used as surfactants in the manufacturing industry. Although it is reported to have estrogenic activity and to be transferred from the mother to the embryo, no data are available on its effects on the development of the human placenta. In the present study, we investigated estrogen receptors' (ERs) expression in the first trimester human placenta. Using an in vitro model of chorionic villous explants, we then compared the effects of p-NP and 17beta-estradiol (17beta-E2). Finally, a trophoblast-derived choriocarcinoma cell line, BeWo, was used as a model of trophoblast cell differentiation. Our results showed that the first trimester placenta expresses three ER-alpha isoforms of 67, 46, and 39 kDa and one ER-beta isoform of 55 kDa. Immunohistochemistry revealed the expression of ER-alpha in the villous cytotrophoblast, whereas ER-beta was mainly expressed by the syncytiotrophoblast. Treatment of explant cultures with p-NP (10(-9)M) and 17beta-E2 (10(-9)M) significantly increased beta-hCG secretion and cell apoptosis but did not modify ER expression. After 72 h of exposure, hormone release was significantly higher in p-NP- than 17beta-E2-treated explant cultures. By this time, cleavage of caspase-3 was evident in cultures treated with 17beta-E2 and p-NP. In BeWo cells, a caspase-3 band of 20-16 kDa was evident after 1 h of treatment with p-NP and after 24 h of treatment with 17beta-E2 or forskolin. These findings suggest that the human trophoblast may be highly responsive to p-NP and raise concern about maternal exposure in early gestation.     

PC-407 inhibited proliferation and induced apoptosis in human colon cancer SW-1116 cells

AIM: To study whether PC-407 [4-[5-naphthyl-3- (trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide] inhibits cell viability and induces apoptosis in human colon cancer SW-1116 cells. METHODS: Inhibition of SW-1116 proliferation was measured by MTT assay. Morphological assessment of apoptosis was performed with fluorescence microscope and electron microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry. RESULTS: PC-407 inhibited SW-1116 cell proliferation in a concentration-dependent manner after 3 d of treatment, and the IC50 for PC-407 inhibition of cell number was 16.67±0.17 μmol/L. After incubation of SW-1116 cells with PC-407 20 μmol/L for 24 h, morphological changes of typical apoptosis were observed by AO/EB staining or transmission electron microscopy. Flow cytometry analysis showed that PC-407 induced apoptosis in SW-1116 cells in a time- and concentration-dependant manner. The agarose gel electrophoresis of DN     

Effects of Endocrine Disruptors on Prosobranch Snails (Mollusca: Gastropoda) in the Laboratory. Part II: Triphenyltin as a Xeno-Androgen

In laboratory experiments the effects of suspected endocrine disrupting chemicals on freshwater and marine prosobranch species were analysed. In this second of three publications the responses of the freshwater ramshorn snail Marisa cornuarietis and of two marine prosobranchs (the dogwhelk Nucella lapillus and the netted whelk Hinia reticulata) to the xeno-androgenic model compound triphenyltin (TPT) are presented. Marisa and Nucella were exposed via water (nominal concentrations 5–500 ng TPT-Sn/L) and Hinia via sediments (nominal concentrations 50–500 g TPT-Sn/kg dry wt.) for up to 4 months. Female ramshorn snails but not the two marine species developed imposex in a time and concentration dependent manner (EC₁₀ 4 months: 12.3 ng TPT-Sn/L) with a comparable intensity as described for tributyltin. TPT reduced furthermore the fecundity of Marisa at lower concentrations (EC₁₀ 4 months: 5.59 ng TPT-Sn/L) with a complete inhibition of spawning at nominal concentrations 250 ng TPT-Sn/L (mean measured ±SD: 163±97.0 ng TPT-Sn/L). The extension of the pallial sex organs (penis with accessory structures and prostate gland) of male ramshorn snails and dogwhelks were reduced by up to 25% compared to the control but not in netted whelks. Histopathological analyses for M. cornuarietis and H. reticulata provide evidence for a marked impairment of spermatogenesis (both species) and oogenesis (only netted whelks). The test compound induced a highly significant and concentration independent increase in the incidence of hyperplasia on gills, osphradia and other organs in the mantle cavity of N. lapillus indicating a carcinogenic potential of TPT. The results show that prosobranchs are sensitive to endocrine disruption at environmentally relevant concentrations of TPT. Also, M. cornuarietis is a promising candidate for a future organismic invertebrate system to identify endocrine-mimetic test compounds.     

生存素反义寡核苷酸诱导HL-60细胞凋亡的实验研究

目的 :探讨特异性靶向生存素的反义寡核苷酸 (ASODN)对急性早幼粒细胞系HL 6 0细胞凋亡的影响。方法 :应用四唑盐 (MTT)比色法分析细胞增殖 ;倒置显微镜观察细胞形态学变化 ;原位末端标记(TUNEL)法检测细胞凋亡指数 (AI) ;流式细胞术(FCM)定量检测细胞凋亡 ;RT PCR半定量检测生存素mRNA的表达。结果 :5～ 2 0 μmol·L-1生存素ASODN对HL 6 0细胞增殖均有抑制作用 ,且呈时间和剂量依赖性。ASODN组凋亡率和生存素基因表达抑制率明显高于对照组。结论 :生存素ASODN能有效抑制HL 6 0细胞生存素mRNA的表达并诱导细胞凋亡 ,表明生存素对维持肿瘤细胞的增殖具有非常重要的作用。