抑制Survivin基因对人卵巢癌SKOV3细胞增殖和凋亡的影响

目的:探讨应用RNAi技术沉默Survivin基因对人卵巢癌SKOV3细胞的影响。方法:构建Survivin基因shRNA真核表达载体,转染人卵巢癌SKOV3细胞。RT-PCR及Western blot检测Survivin基因的表达,MTT实验、流式细胞仪检测细胞增殖、凋亡的变化。结果:siRNA实验组细胞Survivin蛋白及mRNA表达水平明显下降,细胞增殖能力显著降低,细胞凋亡率显著升高。结论:应用RNAi技术沉默Survivin基因可以降低卵巢癌SKOV3细胞Survivin基因的表达,进而抑制肿瘤细胞的生长、增殖并诱导细胞凋亡。因此,Survivin基因可能成为抗肿瘤治疗的潜在靶点。     

紫檀芪对卵巢癌SKOV3细胞凋亡和糖酵解的影响及机制研究

目的 探讨紫檀芪对卵巢癌SKOV3细胞凋亡和糖酵解的影响及机制研究。方法 分别采用0、25、50、100、150 μmol/L的紫檀芪处理SKOV3细胞24、48、72 h,用CCK-8检测紫檀芪对SKOV3细胞增殖的影响,根据CCK-8结果选择后续实验组紫檀芪浓度;采用流式细胞术检测紫檀芪对细胞凋亡的影响;采用葡萄糖氧化酶法和化学比色法检测紫檀芪对SKOV3细胞葡萄糖消耗和乳酸生成的影响;Western blot法检测信号转导与转录激活因子3(STAT3)、磷酸化的STAT3(p-STAT3)、己糖激酶2(HK2)蛋白的表达;qRT-PCR法检测葡萄糖转运蛋白1(GLUT1)、M2型丙酮酸激酶(PKM2)mRNA的表达。结果 CCK-8结果显示,紫檀芪对SKOV3细胞的增殖有抑制作用,且呈时间剂量依赖性,根据CCK-8结果,选择100 μmol/L紫檀芪处理组作为后续实验组,0 μmol/L为对照组;流式细胞术结果显示,紫檀芪可明显促进SKOV3的凋亡,浓度越大,凋亡作用越明显,差异有统计学意义(P＜0.05)。此外,100 μmol/L组紫檀芪作用SKOV3细胞后,葡萄糖消耗和乳酸生成水平均较0 μmol/L组降低(P＜0.01)。Western blot结果显示,与0 μmol/L组相比,100 μmol/L组p-STAT3、HK2蛋白的表达明显降低(P＜0.001)。qRT-PCR结果显示,100 μmol/L组GLUT1、PKM2 mRNA的表达水平也较0 μmol/L组降低(P＜0.01)。结论 紫檀芪可抑制SKOV3细胞的增殖,促进凋亡,并可能通过STAT3/HK2途径抑制卵巢癌的糖酵解。     

低频超声对卵巢癌COC1细胞增殖和凋亡的影响

 目的　观察低频超声对人卵巢癌COC1体外生长的影响。方法　以低频超声 (频率 0 .2 4MHz,剂量 5 .12W /cm2 × 5s)辐照COC1细胞 ,采用MTT观察超声对COC1细胞增殖的抑制作用 ;流式细胞仪(FCM)、TUNEL法和光、电镜检测细胞凋亡。结果　超声波能够显著抑制卵巢癌COC1细胞增殖 ,辐照后 12h、2 4h、36h和 4 8h ,其MTT光吸收值与对照组相比显著降低 ;FCM检测超声辐照后 6h的COC1细胞凋亡率达 (2 5 .5 7± 4 .6 8) % ;同时TUNEL法观察到棕褐色着染的凋亡细胞 ;相差显微镜及电镜下可见凋亡小体形成等典型形态学改变。结论　低频超声辐照对人卵巢癌COC1细胞体外生长具有明显的抑制作用 ,并可诱导其凋亡。     

伏立诺他对卵巢癌SKOV3细胞增殖和凋亡的影响及机制研究

目的 探讨伏立诺他对卵巢癌SKOV3细胞增殖和凋亡的影响及机制研究。方法 以人卵巢癌SKOV3细胞为研究对象,随机分为空白组、伏立诺他高剂量组、中剂量组和低剂量组共4组,采用CCK-8法测定SKOV3细胞增殖情况,Western blot检测SKOV3细胞H3K4ac和H3K27ac蛋白表达情况,Real-time PCR检测SKOV3细胞Bax mRNA、Bcl-2 mRNA、p53 mRNA、CyclinD1 mRNA表达。结果 CCK-8法测定结果显示,与空白组比较,伏立诺他低、中、高剂量组增殖明显降低,差异具有统计学意义(P＜0.05)。RT-PCR结果显示,与空白组比较,伏立诺他低、中、高剂量组SKOV3细胞中Bcl-2 mRNA和CyclinD1mRNA表达明显降低,Bax mRNA和p53 mRNA表达明显升高。Western blot结果显示与空白组比较,伏立诺他低、中、高剂量组SKOV3细胞中H3K4ac和H3K27ac蛋白表达明显增加。结论 伏立诺他抑制SKOV3细胞增殖,促进凋亡,其机制可能与抑制组蛋白去乙酰化酶活性,激活p53凋亡通路有关。     

重组人抗缪勒氏管激素对卵巢癌SKOV3细胞增殖和凋亡的影响

目的:探讨重组人抗缪勒氏管激素（AMH）对人卵巢癌SKOV3细胞的生长抑制作用。方法:采用浓度为25、50、100、200、400 nmol/L重组人AMH处理对数生长期的SKOV3细胞,并设阿霉素（DOX）组为阳性对照组,同时设3个阴性对照孔（DOX和AMH浓度0 nmol/L）,作用72小时后通过CCK-8法检测SKOV3细胞增殖抑制率;采用重组人AMH 100 nmol/L作用于SKOV3细胞48小时后使用Annexin-V和PI染色,并设1 nmol/L DOX为阳性对照组,同时设3个阴性对照孔（DOX和AMH浓度0 nmol/L）,通过流式细胞仪检测AMH对SKOV3细胞的促凋亡作用。结果:浓度100、200、400 nmol/L 的AMH对SKOV3细胞增殖抑制率分别为50.6%、64.5%、78.3%,与阴性对照组相比有统计学差异(P<0.01),且随AMH浓度加大,抑制效果明显提高;流式细胞仪检测AMH组SKOV3细胞总凋亡率为(30.6±2.82)%,与阴性对照组比较,细胞凋亡率明显升高,有统计学差异（P<0.01）。结论:重组人AMH在体外能够明显抑制人卵巢癌SKOV3细胞的增殖,促进其凋亡。     

舒林酸对卵巢癌细胞株A2780、SKOV3的凋亡作用

 舒林酸;卵巢癌;细胞凋亡;凋亡相关基因     

Genistein抑制人卵巢癌细胞系SKOV3增殖和诱导凋亡发生的研究

背景与目的:许多研究表明 Genistein对多种肿瘤细胞有抑制作用,但有关 Genistein对卵巢癌细胞作用的报道很少.本研究旨在通过观测 Genistein对人卵巢癌细胞系 SKOV3的抑制增殖和诱导凋亡作用,探讨其抗癌作用的机理.方法:应用四甲基偶氮唑盐( MTT)法检测不同浓度 Genistein对 SKOV3的生长抑制作用;吖啶橙 /溴乙锭( AO/EB)荧光染色法及电镜观察凋亡细胞及凋亡小体;流式细胞仪分析细胞周期及凋亡率;琼脂糖凝胶电泳检测凋亡特征性 DNA梯形带;免疫细胞化学法及 RT- PCR法分别检测细胞增殖凋亡调控相关蛋白及其 mRNA的表达。结果： Genistein对 SKOV3细胞的增殖抑制作用呈时间及浓度依赖性, 20 μ mol/L和 40 μ mol/L Genistein作用 72 h后,细胞的生长抑制率达 72. 07％和 74. 93％。 20 μ mol/L Genistein作用 SKOV3细胞 48 h后出现细胞周期的 G2/M期阻滞。荧光显微镜和电镜均观察到用药后凋亡细胞典型的形态学特征。细胞凋亡率以 Genistein 20 μ mol/L组最高,达 23. 7％。凝胶电泳观察到特征性 DNA梯形带。 20 μ mol/L Genistein作用 SKOV3 48 h后, bcl- 2基因表达水平降低,而 p21WAF1/CIP1和 bax基因表达增加（ P< 0. 05）; PCNA、Bcl- 2及 cyclin B1蛋白表达水平降低, Bax、p21WAF1/CIP1蛋白表达增加（ P< 0. 01）。结论： Genistein可抑制卵巢癌细胞系 SKOV3的增殖并诱导其发生凋亡, Genistein可能通过上调 p21WAF1/CIP1基因及蛋白水平、下调 cyclin B1及 PCNA蛋白表达水平抑制增殖;通过下调 bcl- 2基因及蛋白表达、上调 bax基因及蛋白表达诱导卵巢癌细胞凋亡.     

乳源免疫调节肽对人卵巢癌SKOV3细胞生长及凋亡的影响

目的:探讨乳源免疫调节肽对人卵巢浆液性乳头状囊腺癌SKOV3细胞生长和凋亡的影响.方法:不同浓度乳源免疫调节肽对体外培养的SKOV3细胞作用24、48、72及96 h后,应用MTT法检测细胞的生长抑制情况,采用HE染色、透射电子显微镜观察及FCM分析细胞周期及细胞凋亡情况.结果:乳源免疫调节肽对SKOV3细胞有增殖抑制作用,与对照组相比差异有统计学意义(P＜0.05),且增殖抑制作用具有时间和浓度依赖性. HE染色和电子显微镜下均观察到用药后出现凋亡细胞的形态学特征.FCM检测发现,乳源免疫调节肽作用后细胞的凋亡率与对照组相比,差异有统计学意义(P＜0.05),其中3×10-4g/L PGPIPN(Pro-Gly-Pro-Zle-Pro-Asn)组的细胞凋亡率最高,达29.4%.结论:乳源免疫调节肽可以抑制体外培养的人卵巢浆液性乳头状囊腺癌SKOV3细胞的增殖,并可诱导其凋亡.     

二氢青蒿素对卵巢癌SKOV3细胞的抑制作用

目的 研究二氢青蒿素（dihydroartemisinin,DHA）诱导卵巢癌SKOV3细胞凋亡。方法 用MTT法检测DHA对SKOV3细胞的增殖抑制作用,流式细胞仪（FCM）测其凋亡率,反转录-聚合酶链反应（RT-PCR）检测Bcl-2、Bax的表达。结果 双氢青蒿素对卵巢癌SKOV3细胞有明显的增殖抑制效应,呈明显的时间-剂量依赖性。DHA处理SKOV3细胞后,24、28、72 h的IC50值分别为117、35.677、23.382 μmol/L。二氢青蒿素可以抑制SKOV3细胞增殖,促进其凋亡,下调Bcl-2基因的表达,增加Bax基因的表达,并有时间、浓度依赖性。结论 二氢青蒿素对人卵巢癌SKOV3细胞有体外抑制作用, 可能通过下调Bcl-2、增加Bax基因的表达来促进细胞的凋亡。     

米非司酮对耐药卵巢癌细胞增殖、凋亡及其对顺铂敏感性的影响

背景与目的:米非司酮是有效的孕酮受体拮抗剂。研究发现,米非司酮对体内外卵巢癌细胞均具有生长抑制作用,但其机制尚不清楚。本研究的目的在于探讨米非司酮对耐药卵巢癌细胞增殖、凋亡及对顺铂(DDP)敏感性的影响和机制,为临床应用米非司酮治疗难治性卵巢癌提供实验依据。方法:体外培养耐药卵巢癌细王刚,等.米非司酮对耐药卵巢癌细胞株SK-OV-3,采用MTT法检测单用米非司酮及合用顺铂时对SK-OV-3细胞增殖的影响,分析米非司酮与顺铂在抑制耐药卵巢癌细胞增殖中的相互作用。采用终末脱氧核糖核苷酸转移酶介导的原位缺口末端标记法和流式细胞术,分析米非司酮及米非司酮联合顺铂对卵巢癌细胞周期和凋亡的影响。结果:实验所选各种浓度米非司酮对SK-OV-3细胞均表现出一定程度的生长抑制作用,并呈现浓度依赖性。当顺铂浓度为1.25μg/ml或2.5μg/ml,合用米非司酮浓度为20μg/ml、10μg/ml、5μg/ml、2.5μg/ml、1.25μg/ml或0.625μg/ml时,能显著抑制SK-OV-3细胞的增殖,并显示出两种药物的协同作用(q>1.15)。当顺铂浓度为0.625μg/ml或5.0μg/ml时,仅表现为两种药物的相加作用(0.85相似文献   

Silencing of Rac3 Inhibits Proliferation and Induces Apoptosis of Human Lung Cancer Cells

Background: Rac3, a member of the Rac family of small guanosine triphosphatases (GTPases), regulatesa variety of cell functions, including the organization of the cytoskeleton, cell migration, and invasion.Overexpression of Rac3 has been reported in several human cancers. However, the role of Rac3 in lung cancer(LC) has not been determined in detail. The purpose of this study was to investigate the effect of silencing of Rac3expression in human LC cells and the consequences for cell survival. Materials and Methods: Lentivirus smallhairpin RNA (shRNA) interference techniques were utilized to knock down the Rac3 gene. Gene and proteinexpression was quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting.LC cell apoptosis was examined by annexin V-APC /propidium iodide staining. Results: Efficient silencing ofRac3 strongly inhibited A549 cell proliferation and colony formation ability, and significantly decreased tumorgrowth. Moreover, flow cytometry analysis showed that knockdown of Rac3 led to G2/M phase cell cycle arrestas well as an excess accumulation of cells in the G1 and S phase. Conclusions: Thus, functional analysis usingshRNAs revealed a critical role for Rac3 in the tumor growth of LC cells. shRNA silencing of Rac3 could providean effective strategy to treat LC.     

HDAC6 siRNA Inhibits Proliferation and Induces Apoptosis of HeLa Cells and its Related Molecular Mechanism

Objective: To investigate the effects of histone deacetylase 6 (HDAC6) siRNA on cell proliferation and cellapoptosis of the HeLa cervical carcinoma cell line and the molecular mechanisms involved. Methods: Divisionwas into three groups: A, the untreated group; B, the control siRNA group; and C, the HDAC6 siRNA group.Lipofectamine 2000 was used for siRNA transfection, and Western blot analysis was used to determine the proteinlevels. Cell proliferation and apoptosis were characterized using a CCK-8 assay and flow cytometry, respectively.Results: HDAC6 protein expression in the HDAC6 siRNA-transfection group was significantly lower (P < 0.05)than in the untreated and control siRNA groups. The CCK-8 kit results demonstrated that the proliferation ofHeLa cells was clearly inhibited in the HDAC6 siRNA transfection group (P < 0.05). In addition, flow cytometryrevealed that the early apoptotic rate (26.0% ± 0.87%) was significantly elevated (P < 0.05) as compared withthe untreated group (10.6% ± 1.19%) and control siRNA group (8.61% ± 0.98%). Furthermore, Western blotanalysis indicated that bcl-2 protein expression in the HDAC6 siRNA–transfection group was down-regulated,whereas the expression of p21 and bax was up-regulated. Conclusion: HDAC6 plays an essential role in theoccurrence and development of cervical carcinoma, and the down-regulation of HDAC6 expression may beuseful molecular therapeutic method.     

Ginsenoside-Rh2 Inhibits Proliferation and Induces Apoptosis of Human Gastric Cancer SGC-7901 Side Population Cells

Objectives: To observed the effects of ginsenoside -Rh2 (GS-Rh2) on proliferation and apoptosis of side population (SP) human gastric cancer SGC-7901 cells. Materials and Methods: SGC-7901 SP and Non-SP cells were sorted by flow cytometry and assessed using the cck-8 method. Expression of apoptosis-related proteins Bax and Bcl-2 of SP before and after the intervention was determined by Western-blotting. Results: It was found that the proliferation of SP was significantly faster than that of NSP (<0.05). In addition, GS-Rh2 inhibited proliferation of gastric cancer SP cells, induced cell cycle arrest and cell apoptosis, and changed the expression of BAX/Bcl-2 proteins in a time-dependent and concentration-dependent manner (<0.05). Conclusions: With increase of GS-Rh2 dose, GS-Rh2 gradually inhibit the proliferation of SGC-7901 SP cells, which have high proliferation rate, through G1/G0 phase arrest, followed by apoptosis which involves the up-regulation of Bax and the down-regulation of Bcl-2.     

小分子化合物PHA-767491对白血病K562细胞抑制增殖和诱导凋亡的作用

目的 探讨PHA-767491对白血病细胞K562的抑制能力及分子机制。方法 采用不同浓度的PHA-767491作用于K562细胞,利用MTT法在不同的时间点检测细胞存活率,RT-PCR及Western blot检测PHA-767491诱导的凋亡信号途径,hochest染色及流式细胞术检测K562的凋亡水平。结果 PHA-767491显著抑制K562细胞的增殖,进一步结果表明PHA-767491可以显著下调抗凋亡蛋白Xiap和Mcl-1蛋白的表达水平,诱导K562细胞通过Caspase途径进入凋亡。结论 PHA-767491可以通过诱导K562细胞凋亡而显著抑制其增殖,为今后的白血病治疗提供了一种新的备选药物。     

小檗碱通过调控IGF-1R信号通路抑制食管癌细胞增殖和诱导细胞凋亡的实验

目的 检测小檗碱对食管癌细胞增殖、周期和凋亡的影响,并探讨其作用机制。 方法 MTT法检测细胞的增殖抑制作用;碘化丙啶（Propidium iodide, PI）染色法检测细胞周期进程;Annexin V-FITC和PI双染法结合流式细胞技术检测小檗碱的凋亡诱导作用;Western blot技术检测小檗碱对胰岛素样生长因子1受体（IGF-1R）的活化及下游主要信号分子AKT和p44/42MAPK（ERK）磷酸化水平的影响。结果 小檗碱在体外可浓度依赖性地抑制四种食管癌细胞系的增殖,且其IC50值与细胞表面IGF-1R的表达水平呈负相关。小檗碱可使细胞阻滞在G₂/M期。细胞凋亡实验结果显示,小檗碱可浓度依赖性地诱导食管癌细胞发生凋亡,并且在相同浓度下KYSE450细胞（IGF-1R高表达）的凋亡率显著高于KYSE150细胞（IGF-1R低表达）。Western blot结果显示,小檗碱处理可显著抑制IGF-1R的磷酸化,并抑制AKT和ERK的活化,且抑制作用随着小檗碱浓度的增加而增强。结论 小檗碱可通过对IGF-1R及其介导的下游信号通路的调控来发挥抑制食管癌细胞增殖、阻断细胞周期进程以及诱导食管癌细胞凋亡的作用。     

MicroRNA-497 Suppresses Proliferation and Induces Apoptosis in Prostate Cancer Cells

MicroRNAs (miRNAs) are a class of endogenously expressed small, non-coding, single-stranded RNAsthat negatively regulate gene expression, mainly by binding to 3’- untranslated regions (3’UTR) of their targetmessenger RNAs (mRNAs), which cause blocks of translation and/or mRNA cleavage. Recently, miRNAprofilingstudies demonstrated the microRNA-497 (miR-497) level to be down-regulated in all prostate carcinomascompared with BPH samples. The purpose of this study was to investigate the potential role of miR-497 inhuman prostate cancer. Proliferation, cell cycle and apoptosis assays were conducted to explore the potentialfunction of miR-497 in human prostate cancer cells. Results showed that miR-497 suppressed cellular growth andinitiated G0/G1 phase arrest of LNCaP and PC-3 cells. We also observed that miR-497 increased the percentageof apoptotic cells by increasing caspase-3/7 activity. Taken together, our results demonstrated that miR-497 caninhibit growth and induce apoptosis by caspase-3 activation in prostate cancer cells, which suggest its use as apotential therapeutic target in the future.     

Suppression of Cellular Apoptosis Susceptibility (CSE1L)Inhibits Proliferation and Induces Apoptosis in ColorectalCancer Cells

The cellular apoptosis susceptibility (CSE1L) gene has been demonstrated to regulate multiple cellularmechanisms including the mitotic spindle check point as well as proliferation and apoptosis. However, theimportance of CSE1L in human colon cancer is largely unknown. In the present study, we examined expressionlevels of CSE1L mRNA by semiquantitative RT-PCR. A lentivirus-mediated small interfering RNA (siRNA)was used to knock down CSE1L expression in the human colon cancer cell line RKO. Changes in CSE1L targetgene expression were determined by RT-PCR. Cell proliferation was examined by a high content screeningassay. In vitro tumorigenesis was measured by colony-formation assay. Cell cycle distribution and apoptosiswere detected by flow cytometric analysis. We found CSE1L mRNA to be expressed in human colon cancer cells.Using a lentivirus based RNAi approach, CSE1L expression was significantly inhibited in RKO cells, causingcell cycle arrest in the G2/M and S phases and a delay in cell proliferation, as well as induction of apoptosisand an inhibition of colony growth capacity. Collectively, the results suggest that silencing of CSE1L may be apotential therapeutic approach for colon cancer.     

Silibinin Inhibits Proliferation,Induces Apoptosis and Causes Cell Cycle Arrest in Human Gastric Cancer MGC803 Cells Via STAT3 Pathway Inhibition

Background: To investigate the effect of silibinin on proliferation and apoptosis in human gastric cancer cell line MGC803 and its possible mechanisms. Materials and Methods: Human gastric cancer cell line MGC803 cells were treated with various concentration of silibinin. Cellular viability was assessed by CCK-8 assayandapoptosis and cell cycle distribution by flow cytometry. Protein expression and mRNA of STAT3, and cell cycle and apoptosis regulated genes were detected by Western blotting and real-time polymerase chain reaction, respectively. Results: Silibinin inhibits growth of MGC803 cells in a dose- and time-dependent manner. Silibinin effectively induces apoptosis of MGC803 cells and arrests MGC803 cells in the G2/M phase of the cell cycle, while decreasing the protein expression of p-STAT3, and of STAT3 downstream target genes including Mcl-1, Bcl-xL, survivin at both protein and mRNA levels. In addition, silibinin caused an increase in caspase 3 and caspase 9 protein as well as mRNA levels. Silibinin caused G2/M phage arrest accompanied by a decrease inCDK1 and Cyclin B1 at protein and mRNA levels.. Conclusions: These results suggest that silibinin inhibits the proliferation of MGC803 cells, and it induces apoptosis and causes cell cycle arrest by down-regulating CDK1, cyclinB1, survivin, Bcl-xl, Mcl-1 and activating caspase 3 and caspase 9, potentially via the STAT3 pathway.     

洛铂诱导顺铂耐药卵巢癌SKOV3/ DDP细胞的凋亡

 目的 探讨洛铂体外诱导人卵巢癌顺铂耐药SKOV3／DDP细胞凋亡及其机制。方法 采用四甲基偶氮唑盐（MTT）法测定不同浓度和时间洛铂对SKOV3／DDP细胞的生长抑制作用，并与其亲本细胞SKOV3（敏感株）相比较；进一步通过光镜及透射电镜观察SKOV3／DDP细胞的形态学改变；并采用流式细胞仪检测细胞周期变化。结果 洛铂作用SK0V3／DDP细胞的增殖呈时间剂量依赖模式受到抑制，与SKOV3细胞相比，差异无统计学意义（P〉0．05）；一定浓度的SKOV3／DDP作用一定时间，可诱导SKOV3／DDP细胞发生凋亡；光镜和透射电镜下可见典型的凋亡细胞形态学改变；流式细胞技术分析发现洛铂作用首先引起细胞周期分布的改变，随用药时间的延长，凋亡率逐渐升高。结论 洛铂通过诱导凋亡而抑制体外培养的SKOV3／DDP细胞生长，其机制可能与细胞G0／G1期阻滞有关。