槲皮素抑制离体大鼠肝星状细胞增殖

目的：研究酪氨酸蛋白激酶抑制剂槲皮素对肝星状细胞增殖的影响。方法：用链酶蛋白酶和胶原酶原位灌流消化正常大鼠肝脏，Nycodenz密度梯度离心分离肝星状细胞，采用MTT比色法，流式细胞术检测细胞增殖水平。结果：槲皮素处理组可显著地抑制肝星状细胞的增殖并呈剂量依赖性关系；可使G0／G1期细胞增多，S期细胞减少。结论：槲皮素可明显抑制肝星状细胞增殖。     

XAV939对卵巢癌细胞增殖的抑制作用

目的观察XAV939对卵巢癌细胞株OVCAR3增殖的作用。方法采用MTT法检测XAV939对卵巢癌细胞增殖的抑制作用,用流式细胞技术检测卵巢癌细胞株OVCAR3细胞周期和凋亡,Western blotting法检测Cyclin D1、Bcl-2的表达以及PARP剪切片段。结果 XAV939对卵巢癌细胞株增殖的抑制呈现浓度依赖性和时间依赖性,细胞周期检测表现为G1期阻滞,凋亡检测显示凋亡率增加。不同浓度的XAV939处理组的卵巢癌细胞Cyclin D1、Bcl-2蛋白表达水平下降,PARP剪切片段增多。结论 XAV939可以抑制卵巢癌细胞株OVCAR3的增殖。     

PIAS3敲低对前列腺癌细胞增殖和凋亡的影响

目的研究PIAS3敲低对人前列腺癌细胞系DU145细胞增殖、细胞周期和凋亡的影响。方法构建PIAS3shRNA表达质粒pSilencer4.1/PIAS3,转染DU145细胞。MTT法检测细胞增殖,流式细胞仪分析细胞周期和凋亡。结果测序证实PIAS3shRNA表达质粒构建成功。转染后DU145细胞PIAS3蛋白表达明显下调。MTT分析显示PIAS3敲低促进细胞增殖,并存在剂量-效应关系。流式细胞术分析显示:PIAS3敲低后S期细胞比例增加,G0/G1期细胞比例减少,凋亡细胞比例减少。结论PIAS3敲低促进体外前列腺癌细胞增殖,抑制其凋亡。     

Nucleostemin基因在姜黄素抑制胃癌SGC-7901细胞增殖中的意义

目的探讨NS基因在姜黄素抑制胃癌SGC-7901细胞株增殖和诱导凋亡中的作用。方法不同浓度姜黄素作用于胃癌SGC-7901细胞株,利用MTT法检测细胞增殖抑制率,RT-PCR法检测姜黄素作用前后NS基因表达量的变化,流式细胞仪法检测细胞周期和细胞凋亡情况。结果姜黄素能显著抑制体外培养的胃癌SGC-7901细胞株增殖,并呈量效和时效关系;流式细胞术显示G0/G1期细胞增加,S期细胞减少,检出亚二倍体凋亡峰;RT-PCR显示NS基因表达量下降。结论姜黄素显著抑制胃癌SGC-7901细胞株增殖,并促进凋亡,其发生可能与NS基因表达下降有关。     

TRIM19通过调控p53信号促进卵巢癌进展的作用及机制研究

目的:探讨TRIM19在卵巢癌进展中的作用及其分子机制。方法:qPCR检测人卵巢表皮细胞IOSE80、卵巢癌细胞SKOV-3、A2780、OVCAR3中TRIM19表达水平;以siRNA敲减SKOV-3细胞中TRIM19基因表达;以CCK-8方法、Transwell实验、流式细胞术分别检测细胞增殖能力、迁移行为及凋亡率;以RT-qPCR技术检测信号通路分子表达。结果:在卵巢癌细胞SKOV-3、A2780、OVCAR3中,TRIM19表达量较正常表皮细胞IOSE80中明显增强。当TRIM19基因在卵巢癌SKOV-3细胞中被成功敲减后,细胞增殖能力降低74.2%,侵袭能力降低70.0%,凋亡率增加约30倍。在分子水平,敲减TRIM19后,p53、p21、CyclinD、CDK2、Bax基因表达显著增加;同时敲减TRIM19与p53后,p53、p21、CyclinD、CDK2、Bax基因表达略有提高。结论:沉默TRIM19基因抑制卵巢癌细胞增殖、侵袭,诱导凋亡,并且激活p53依赖的凋亡信号,TRIM19促进卵巢癌进程。TRIM19具有成为卵巢癌治疗的新靶点潜力。     

三氧化二砷诱导卵巢癌HO8910细胞凋亡

目的探讨三氧化二砷对卵巢癌细胞株HO8910的抑制增殖效应及凋亡诱导作用。方法用三氧化二砷处理HO8910细胞,采用MTT法检测药物对细胞的抑制作用,倒置显微镜和电镜观察细胞形态学改变,琼脂糖凝胶电泳观察DNA降解,以及应用流式细胞仪观察细胞凋亡过程中细胞周期的变化。结果在三氧化二砷作用下,HO8910细胞呈凋亡改变,DNA琼脂糖凝胶电泳呈典型的凋亡特征。细胞凋亡的同时,细胞周期发生特定的改变,亚G1峰出现,S+G2~M期细胞所占比例组明显增高。结论三氧化二砷能诱导卵巢癌细胞凋亡,抑制卵巢癌细胞增殖。     

槲皮素抗结肠癌细胞SW480生长的作用机制研究

【摘要】　目的　研究槲皮素抗结肠癌细胞SW480 生长的作用, 并探索其可能的作用机制。方法　通过CCK-8 法来检测SW480 细胞在槲皮素处理下的增殖变化; 运用HE 染色、电镜观察和流式细胞术方法检测SW480 细胞在槲皮素处理下的凋亡变化; 运用Transwell 检测槲皮素处理情况下SW480 细胞在体外的粘附、运动和侵袭能力变化; 运用明胶酶谱分析法检测槲皮素处理情况下SW480 细胞基质金属蛋白酶分泌能力的变化。结果　CCK-8 法检测显示槲皮素处理使SW480 细胞受到了显著的增殖抑制, 且具有剂量和时间依赖 性; HE 染色、流式细胞术及电镜分析发现槲皮素明显诱导细胞凋亡, 并将SW480 细胞周围阻滞于G2 / M期; 此外, 槲皮素也能够使结肠癌SW480 细胞的体外侵袭和运动能力受到抑制, 且该抑制作用呈剂量依赖性关系; 明胶酶谱法显示槲皮素处理后, SW480 细胞对金属蛋白酶2 (matrix metalloproteinase-2, MMP-2)和金属蛋白酶9 (matrix metalloproteinase-9, MMP-9) 的分泌均下降。结论　槲皮素对人结肠癌SW480 细胞的增殖有明显的抑制作用, 并能促进SW480 细胞发生凋亡。槲皮素对结肠癌SW480 细胞的迁移及侵袭能力均表现出抑制作用, 并且呈剂量依赖性, 这可能和槲皮素能够抑制MMP-2 及MMP-9 的分泌有关。     

抗HER-2嵌合抗体chA21对SKOV3细胞增殖的抑制及诱导其凋亡的作用

目的：探讨抗HER-2嵌合抗体chA21在体外抑制高表达HER-2的人卵巢癌SKOV3细胞增殖并诱导其凋亡的作用。方法：采用MTT比色法、HE染色、透射电镜、流式细胞术及TUNEL染色法等观察和检测chA21对人卵巢癌细胞SKOV3增殖抑制和凋亡的诱导作用：结果：chA21（0．2mg／L～5．4mg／L）可显著抑制SKOV3细胞增殖并诱导其凋亡．其作用呈剂量和时间依赖性：结论：chA21在体外可显著抑制SKOV3细胞的增殖，诱导凋亡可能是其主要的作用途径，     

苦参素诱导卵巢癌HO8910细胞凋亡的实验研究

目的探讨苦参素对卵巢癌细胞株HO8910的抑制增殖效应及凋亡诱导作用。方法用不同浓度苦参素处理HO8910细胞,采用MTT法检测药物对细胞的抑制作用,倒置显微镜和电镜观察细胞形态学改变;以及应用流式细胞仪观察细胞凋亡过程中细胞周期的变化。结果随着药物浓度的升高,作用时间的延长,苦参素对HO8910细胞的生长抑制率逐渐增高;在苦参素作用下,HO8910细胞呈凋亡改变,出现凋亡小体。细胞凋亡的同时,细胞周期发生特定的改变,亚G1峰出现,S+G2～M期细胞所占比例组明显增高。结论苦参素能诱导卵巢癌细胞凋亡,抑制卵巢癌细胞增殖。     

ORP-150基因沉默对SKOV-3细胞增殖、迁移及侵袭的影响

目的:探讨沉默ORP-150基因对体外培养卵巢癌细胞SKOV-3的增殖、转移及侵袭的影响并初步探讨其可能作用机制.方法:运用慢病毒包装shRNA方法沉默SKOV-3细胞的ORP-150基因;研究分为3组:实验组(ORP-150 shRNA)、阴性对照组(NC shRNA)和空白对照组(CON);采用qRT-PCR和Western印迹法检测基因敲减效率,CCK8法、克隆形成试验检测ORP-150沉默对细胞增殖能力的影响,划痕试验、Transwell试验检测SKOV-3细胞侵袭迁移能力的变化,流式细胞仪检测细胞的凋亡情况,Western印迹法探讨ORP-150基因敲减后影响细胞表型改变的作用机制.结果:沉默ORP-150基因可以抑制卵巢癌细胞的增殖能力、克隆形成能力以及跨膜转移扩散能力(P<0.05),但对细胞的侵袭能力无明显影响(P>0.05);敲减ORP-150基因后卵巢癌细胞凋亡明显增加(P<0.05);敲减ORP-150基因后的卵巢癌细胞可能通过Wnt信号通路及Caspase级联反应影响细胞的修复,最终抑制细胞增殖,促进凋亡发生.结论:沉默ORP-150基因可以抑制卵巢癌细胞SKOV-3的增殖和转移,同时促进凋亡发生.这种细胞表型的改变可能是通过下调Wnt信号通路中β-catenin,同时的表达,以及抑制Caspase级联反应从而下调c-myc,cyclin D1和caspase-3蛋白的表达来实现,ORP-150基因沉默可能是卵巢癌治疗的新靶点.     

槲皮素对U937细胞系抑制增殖和诱导凋亡作用的研究

目的 探讨黄酮类化合物槲皮素（Que）对人类单核细胞白血病U937细胞系的抑制增殖和诱导凋亡的作用。方法 应用MTT法检测不同浓度槲皮素对U937细胞的增殖抑制作用；AO／PI荧光染色后倒置荧光显微镜下观察细胞形态学变化；琼脂糖凝胶电泳测定细胞DNA的片段化；应用流式细胞仪检测细胞凋亡率及细胞周期分布。结果槲皮素能明显抑制U937细胞增殖，并存在剂量-效应关系和时间-效应关系；诱导U937细胞出现凋亡所具有的形态学和生化特征；随着槲皮素浓度升高，凋亡细胞和坏死细胞比例增加；将细胞特异性地阻滞在S期，出现凋亡峰。结论 槲皮素能抑制U937细胞增殖，诱导细胞凋亡，并具有细胞周期特异性。     

FTO accelerates ovarian cancer cell growth by promoting proliferation,inhibiting apoptosis,and activating autophagy

ObjectiveFat mass and obesity-associated protein (FTO) is identified as a critical demethylase involved in various physiological processes. Despite efforts have been made to study the biological functions of FTO in certain cancers, the role of FTO in ovarian cancer is largely unknown. In this study, we sought to investigate the function of FTO on proliferation, apoptosis and autophagy of ovarian cancer cells.MethodsQuantitative real-time PCR was performed to detect FTO expression in ovarian tumor tissues and ovarian cancer cell lines OVCAR-3, SKOV-3, COC1, HO-8910 and A2780. SKOV-3 cells were constructed with FTO overexpression and A2780 cells were constructed with FTO knockdown. CCK-8 assay was used to examine cell viability and flow cytometry was used to detect cell apoptosis. Activity assay kits were applied to detect caspase-3 and caspase-9 levels. Western blot was performed to measure the expressions of FTO, PCNA, Bax, Bcl-2, LC3, ATG5, P62, p-AKT and AKT. Stable FTO-overexpression SKOV-3 cells or FTO-depletion A2780 cells were injected subcutaneously into male Balb/c-nu mice. Xenografted tumors were assayed by H&E staining. Immunohistochemistry was subjected to measure FTO and Ki67 expressions.ResultsFTO was up-regulated in ovarian tumor tissues compared with non-cancerous ovarian tissues. FTO overexpression markedly increased viability and autophagy function, but decreased apoptosis of ovarian cancer cells. In addition, FTO overexpression promoted AKT phosphorylation. In contrast, FTO silence showed the opposite effect.ConclusionFTO accelerated ovarian cancer cell growth by promoting proliferation, inhibiting apoptosis, and activating autophagy.     

A novel histone deacetylase inhibitor, Scriptaid, induces growth inhibition, cell cycle arrest and apoptosis in human endometrial cancer and ovarian cancer cells

Histone deacetylase inhibitors (HDACIs) can inhibit cell proliferation, induce cell cycle arrest, and stimulate the apoptosis of cancer cells. We investigated the effects of a novel HDACI, Scriptaid, on the Ishikawa endometrial cancer cell line, SK-OV-3 ovarian cancer cell line, and normal human endometrial epithelial cells. Endometrial and ovarian cancer cells were treated with various concentrations of Scriptaid, and its effect on cell growth, cell cycle, apoptosis, and related measurements was investigated. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays showed that all endometrial and ovarian cancer cell lines were sensitive to the growth inhibitory effect of Scriptaid, although normal endometrial epithelial cells were viable after treatment with the same doses of Scriptaid that induced the growth inhibition of endometrial and ovarian cancer cells. Cell cycle analysis indicated that their exposure to Scriptaid decreased the proportion of cells in the S phase and increased the proportion in the G0/G1 and/or G2/M phases of the cell cycle. Induction of apoptosis was confirmed by annexin V staining of externalized phosphatidylserine and loss of the transmembrane potential of mitochondria. This induction occurred in concert with the altered expression of genes related to cell growth, malignant phenotype, and apoptosis. Furthermore, Scriptaid treatment of these cell lines increased acetylation of H3 and H4 histone tails. These results raise the possibility that Scriptaid may prove particularly effective in the treatment of endometrial and ovarian cancers.     

槲皮素通过抑制c-Jun的表达水平增强5-氟尿嘧啶对胃癌细胞凋亡的诱导活性

目的:探讨槲皮素是否能增强5-氟尿嘧啶对胃癌细胞凋亡的诱导活性并研究其机制。方法:MTT法检测5-氟尿嘧啶在槲皮素辅助治疗下对胃癌细胞系BGC-823的杀伤活性和半抑制浓度(IC50)。免疫共沉淀和Western blot实验检测5-氟尿嘧啶和槲皮素对BGC-823细胞c-Jun和Bcl-x L的表达水平、c-Jun磷酸化水平、caspase-9和caspase-3活化水平以及细胞色素C释放的影响。流式细胞术检测5-氟尿嘧啶在槲皮素辅助下对BGC-823细胞凋亡的影响。结果:槲皮素能明显提高5-氟尿嘧啶对BGC-823细胞的杀伤活性,降低5-氟尿嘧啶对BGC-823细胞的IC50。槲皮素处理明显抑制BGC-823细胞c-Jun的表达,并抑制BGC-823细胞中5-氟尿嘧啶诱导的c-Jun磷酸化及其与ATF2蛋白的相互作用,进而抑制5-氟尿嘧啶诱导的Bcl-x L蛋白上调。转染c-Jun过表达质粒后,槲皮素联合5-氟尿嘧啶对BGC-823细胞的杀伤活性受到显著抑制。同时,槲皮素能显著增强BGC-823细胞中5-氟尿嘧啶诱导的细胞色素C从线粒体中释放和caspase依赖的凋亡。结论:槲皮素可通过c-Jun/ATF2/Bcl-x L途径增强5-氟尿嘧啶对胃癌细胞线粒体途径凋亡的诱导活性。     

Effect of plasmid-mediated stable interferon-γ expression on proliferation and cell death in the SKOV-3 human ovarian cancer cell line

Introduction: High interferon-γ (IFN-γ) expression in tumors has been reported to be a favorable prognostic marker. Continuous exposure of ovarian cancer cells to IFN-γ was previously shown to result in significant growth inhibition and apoptosis. Our goal in this study was to evaluate the effect of plasmid-mediated stable IFN-γ expression on the SKOV-3 human ovarian carcinoma cell line.

Methods: SKOV-3 cells were stably transfected with the pEGFP-IFN-γ plasmid. IFN-γ mRNA was detected by RT-PCR and IFN-γ protein expression was measured by ELISA. Proliferation and cell death in transfected SKOV-3 cells were measured by methyl-thiazolyl tetrazolium (MTT) assay and Hoechst 33258 staining, respectively and compared with untransfected and empty vector-transfected cells.

Results: pEGFP-IFN-γ SKOV-3 cells efficiently expressed and secreted IFN-γ. They exhibited significantly decreased cellular proliferation when compared with control untransfected or empty vector-transfected cells (P?<?0.05). The mode of cell death was primarily apoptosis.

Conclusions: Stable expression of IFN-γ significantly inhibits the proliferation of ovarian carcinoma cells and has the potential to be used in clinical applications to treat ovarian carcinoma in the future.     

EGCG通过调控SIRT1-P53通路诱导人卵巢癌SKOV-3细胞凋亡

 目的: 研究表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)调控人卵巢癌SKOV-3细胞活力和凋亡的分子机制。方法: SKOV-3细胞给予EGCG(0~50 μmol/L)、SIRT1激动剂SRT1720(1 μmol/L)和SIRT1抑制剂EX527(1 μmol/L)处理后,用CCK-8法检测细胞活力,流式细胞术检测细胞凋亡,real-time PCR检测Bax和Bcl-2 mRNA的表达水平;采用SIRT1去乙酰化酶活性检测试剂盒检测SIRT1酶活性;使用Western blot法检测SIRT1、乙酰化P53和P53的蛋白表达变化。结果: 与正常对照组相比,单独给予EGCG或EX527处理之后SKOV-3细胞活力下降,凋亡率增加;SIRT1的酶活性和蛋白表达水平均明显下降;P53的乙酰化水平显著增加。与EGCG组相比,SRT1720预处理组的细胞活力上升,凋亡率下降,Bax/Bcl-2的相对比值及激活型caspase-3的蛋白水平明显下降,并且SIRT1的酶活性和蛋白表达水平显著增加,P53的乙酰化水平下降。结论: EGCG可通过调控SIRT1-P53通路抑制卵巢癌SKOV-3细胞活力并诱导其凋亡。     

穿心莲内酯对卵巢癌细胞株SKOV-3侵袭与凋亡的影响

目的:观察穿心莲内酯对卵巢癌细胞株SKOV-3侵袭与凋亡的影响并探讨初步的作用机制。方法:CCK-8法检测不同浓度(0、5、10、20和40μmol/L)的穿心莲内酯对SKOV-3细胞作用不同时间(12、24、36和48h)后,SKOV-3细胞存活率的变化;Transwell法与TUNEL法分别检测SKOV-3细胞侵袭能力与凋亡能力的变化;Western blot法检测p-PI3K、p-Akt与p-mTOR蛋白水平的变化。结果:CCK-8法检测结果显示,随着浓度增加与培养时间延长,穿心莲内酯对SKOV-3细胞存活率的抑制程度增强;采用20μmol/L穿心莲内酯培养SKOV-3细胞36h后,SKOV-3细胞的侵袭数明显降低,凋亡数明显增加(P0.05),p-PI3K、p-Akt与p-mTOR的蛋白水平明显降低(P0.05)。结论:穿心莲内酯能够抑制卵巢癌细胞SKOV-3的活力与侵袭能力,增强凋亡能力,这可能与抑制PI3K/Akt/mTOR信号通路有关。     

Effects of quercetin, a plant flavonol, on the two-stage transformation in vitro

Quercetin was examined for the effects on the two-stage chemical transformation of BALB/3T3 cells. Quercetin showed initiating action to induce transformation in the cells which were treated with quercetin and subsequently with 0.49 microM 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Both the proportion of dishes with transformed foci and the average number of foci per dish increased with the concentration of quercetin (15-45 microM). However, initiating treatment with quercetin did not induce transformation without subsequent TPA treatment. Quercetin inhibited the promotion caused by 0.49 microM TPA in the transformation initiated by 1.9 microM 2-methylcholanthrene (MCA). The inhibitory effect of 30 microM quercetin was 56% in the number of foci per dish. Thus quercetin was found to have initiating effect on the transformation of BALB/3T3 cells, but to restrain the promotion by TPA.     

Medroxyprogestogen enhances apoptosis of SKOV-3 cells via inhibition of the PI3K/Akt signaling pathway

We sought to assess the effect of progestin on the apoptosis of epithelial ovarian cancer cell line SKOV-3 and via regulation of phosphorylation signaling in.Epithelial ovarian cancer cell line SKOV-3 was treated with medroxyprogestogen,phosphatidylinositol 3-kinase inhibitor LY294002 and vehicle control.Akt,phospho-Akt,Bcl-2 and phospho-Bad proteins were examined by immunoblotting assays.Medroxyprogestogen-induced apoptosis was assessed by MTT assays and Annexin V apoptosis assay.We found no significant difference in Akt and Bad expression in both the medroxyprogestogen groups and the control group.The levels of phospho-Akt,Bcl-2 and phospho-Bad were decreased in all the medroxyprogestogen groups and significantly decreased in the high dose mitogen-activated protein (MAP) group (10 μmol/L).Viability of SKOV-3 was reduced and apparent apoptosis of SKOV-3 cells was observed with increased doses of MAP.The findings suggest that medroxyprogestogen can induce SKOV-3 cell apoptosis by inhibiting Akt phosphorylation.