利用基因芯片技术筛选人卵巢癌耐药差异表达基因

摘 要：［目的］ 比较卵巢上皮癌铂类耐药和非铂类耐药细胞间的基因表达差异,并进行相关基因的筛选和鉴定。［方法］ 采用人类全基因组表达谱芯片对卵巢癌铂类耐药细胞SKOV3/CB和亲本细胞SKOV3进行差异基因的筛选和分析;用半定量逆转录聚合酶联反应(RT-PCR)对部分芯片结果进行核对验证。［结果］ 共分离筛选出差异基因3506个,SKOV3/CB比SKOV3上调的差异基因共有1712个,其中上调10倍以上的共有163个,下调的差异基因共有1794个,其中下调10倍以上的共有70个。SKOV3/CB比SKOV3上调10倍以上的3个差异基因,即ZNF198、ERCC5和BIRC3经半定量RT-PCR验证,表明上述3个基因在耐药细胞株中的表达的确高于敏感细胞株。［结论］ 卵巢癌的多药耐药是一个多种基因参与的复杂过程,ZNF198、ERCC5和BIRC3参与卵巢癌铂类耐药。     

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。     

芬太尼调控LINC00184抑制卵巢癌SKOV3细胞增殖和侵袭

目的:研究芬太尼对卵巢癌SKOV3细胞增殖、凋亡和侵袭等生物学行为的影响,并探讨其作用机制。方法:体外培养人卵巢癌SKOV3细胞,采用CCK-8实验、克隆形成实验检测芬太尼对SKOV3细胞体外增殖的影响;分别采用流式细胞术和Transwell实验检测芬太尼对SKOV3细胞凋亡和侵袭能力的影响;Western blot检测细胞增殖、凋亡及侵袭相关蛋白表达。qRT-PCR检测LINC00184在卵巢癌细胞系和正常卵巢上皮细胞株的表达;构建LINC00184过表达质粒并转染SKOV3细胞,观察上调LINC00184表达对芬太尼处理的SKOV3细胞增殖、凋亡及侵袭的影响。结果:芬太尼在体外呈时间和浓度依赖性抑制SKOV3细胞存活,而不明显影响正常卵巢上皮细胞HOSE的存活率。相比对照组,使用5 ng/mL和10 ng/mL芬太尼处理SKOV3细胞,显著下调LINC00184表达水平,降低体外克隆形成率,抑制Ki67、PCNA蛋白表达（均P＜0.01）;显著增加SKOV3细胞凋亡率,上调Bax蛋白表达,下调Bcl-2蛋白表达（均P＜0.01）;显著降低侵袭细胞数,上调E-cadherin蛋白表达,下调N-cadherin、MMP9蛋白表达（均P＜0.01）。而使用LINC00184过表达质粒上调LINC00184表达则明显削弱芬太尼对SKOV3细胞的上述作用（P＜0.01）。结论:LINC00184在卵巢癌细胞表达上调,并与卵巢癌细胞的增殖、凋亡和侵袭等生物学特性相关;芬太尼通过下调LINC00184表达抑制卵巢癌细胞的增殖和侵袭,并诱导其凋亡。     

MiRNA-210通过PI3/AKT信号通路上调E2F3促进卵巢癌SKOV3细胞增殖作用

摘 要：［目的］ 研究miRNA-210是否通过PI3/AKT信号通路上调E2F3促进卵巢癌SKOV3细胞的增殖作用。［方法］ 收集24例卵巢癌患者的临床组织。QRT-PCR分别检测肿瘤组织和癌旁组织中miRNA-210的表达;Western blot检测肿瘤组织和癌旁组织中AKT、P-AKT、E2F3表达;CCK-8检测三组细胞的增殖能力;平板克隆形成实验检测三组细胞的克隆形成能力;裸鼠皮下荷瘤模型检测三组细胞的成瘤能力。［结果］ 肿瘤组织中miRNA-210的表达高于癌旁组织;肿瘤组织中P-AKT 和E2F3的表达高于癌旁组织;转染结束后,三组细胞中过表达载体miRNA-210转染SKOV3细胞中miRNA-210表达明显高于对照组和空白组,而对照组和空白组之间无差异;CCK-8检测结果显示miRNA-210过表达后促进了SKOV3细胞增殖;平板克隆形成实验结果显示miRNA-210过表达后SKOV3细胞克隆形成能力明显增强;裸鼠皮下荷瘤模型结果显示miRNA-210过表达后SKOV3细胞皮下成瘤能力明显强于对照组和空白组;Western blot结果显示miRNA-210过表达后P-AKT和E2F3表达增加,但是miRNA-210过表达SKOV3细胞系中加入1μmol/L Wortmannin后E2F3表达随之下调。［结论］ MiRNA-210通过PI3/AKT信号通路上调E2F3,从而促进卵巢癌SKOV3细胞增殖作用。     

IFN-α和TGF-β1对卵巢癌细胞SKOV3端粒酶活性的影响

目的：观察IFN－α和TGF－β1对卵巢癌细胞系SKOV3端粒酶活性和凋亡的影响，探讨它们在肿瘤治疗中的作用机制。方法：用IFN－α和KTGF－β1分别处理SKOV3细胞，然后采用端粒酶PCR ELISA方法检测不同处理时间细胞的端粒酶活性表达，同时利用流式细胞仪观察处理前后SKOV3细胞的凋亡情况。用裸鼠体内成瘤实验及免疫组化，检测转染TGF－β1基因对SKOV3细胞生长及端粒酶活性的影响。结果：IFN－α对SKOV3细胞生长有抑制作用，可明显下调SKOV3细胞端粒酶活性；而TGF－β1作用则相反，在体内外对SKOV3细胞生长均有促进作用，可明显上调SKOV3细胞端粒酶活性；IFN－α和TGF－β1均不诱导SKOV3细胞发生凋亡。结论：抑制端粒酶活性是IFN－α抗卵巢癌治疗的一个重要机制；TGF－β1在体内外对SKOV3细胞生长均有促进作用。     

抑制survivin基因对人卵巢癌SKOV3细胞顺铂药物敏感性的影响

目的：探讨应用RNAi技术下调survivin基因对人卵巢癌SKOV3细胞增殖、凋亡及顺铂敏感性的影响。方法：构建survivin基因shRNA真核表达载体,转染人卵巢癌SKOV3细胞。定量PCR和Western blotting观察SKOV3细胞survivinmRNA和蛋白表达的改变;噻唑蓝（Myr）检测细胞增殖活性和药物敏感性;流式细胞仪检测细胞凋亡。结果：SKOV3-siRNA组细胞survivinmRNA及蛋白表达下降,同时细胞增殖能力明显降低,细胞凋亡率显著增加（P〈0．05）。SKOV3-siRNA组细胞对顺铂的化学敏感性升高,顺铂的IC50值降低（P〈0．05）。结论：下调survivin基因表达能够抑制卵巢癌SKOV3细胞增殖能力,诱导细胞凋亡,增强细胞顺铂药物敏感性。因此,survivin基因可能成为抗肿瘤治疗的潜在靶点。     

抑制survivin基因对人卵巢癌SKOV3细胞顺铂药物敏感性的影响(英文)

目的:探讨应用RNA i技术下调survivin基因对人卵巢癌SKOV3细胞增殖、凋亡及顺铂敏感性的影响。方法:构建survivin基因shRNA真核表达载体,转染人卵巢癌SKOV3细胞。定量PCR和Western blotting观察SKOV3细胞survivin mRNA和蛋白表达的改变;噻唑蓝(MTT)检测细胞增殖活性和药物敏感性;流式细胞仪检测细胞凋亡。结果:SKOV3-siRNA组细胞survivin mRNA及蛋白表达下降,同时细胞增殖能力明显降低,细胞凋亡率显著增加(P<0.05)。SKOV3-siRNA组细胞对顺铂的化学敏感性升高,顺铂的IC50值降低(P<0.05)。结论:下调survivin基因表达能够抑制卵巢癌SKOV3细胞增殖能力,诱导细胞凋亡,增强细胞顺铂药物敏感性。因此,survivin基因可能成为抗肿瘤治疗的潜在靶点。     

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.     

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。     

溶血磷脂酸抑制顺铂诱导的卵巢癌细胞凋亡

目的：探讨溶血磷脂酸（lysophosphatidic acid,LPA）对顺铂诱导的卵巢癌细胞凋亡的抑制作用及其作用机制。方法：体外培养卵巢癌细胞株SKOV3,采用MTT法检测LPA对顺铂（cisplatin,DDP）作用后卵巢癌细胞株SKOV3增殖活性的影响,Hoechst33258荧光染色观察凋亡细胞,用FCM法分析细胞周期变化和细胞凋亡率,DNA片段凝胶电泳观察凋亡细胞的DNA“梯状”条带,免疫细胞化学法及RT-PCR法分别检测细胞凋亡相关蛋白及其mRNA的表达。结果：LPA能降低DDP对SKOV3细胞生长的抑制作用,同时增加G0/G1期细胞比例,降低S期细胞比例和凋亡率。Hoechst33258染色检测示LPA作用后凋亡小体明显减少。DNA片段凝胶电泳示LPA作用后不产生明显的凋亡片段。10μmol/L的LPA作用SKOV3细胞48h后,bcl-2基因及其蛋白表达水平升高,而bax基因及其蛋白表达水平降低（P〈0.01）。结论：LPA可通过上调bcl-2基因及蛋白表达,下调bax基因及蛋白表达,抑制DDP诱导的卵巢癌细胞凋亡。因此,针对LPA的治疗有望提高DDP疗效,改善卵巢癌患者的预后。     

Bacteria and cancer--antagonisms and benefits

There is considerable historical and recent evidence concerning the antagonisms between acute bacterial infections or their toxins and cancer and allied diseases. These data provide renewed incentives to undertake clinical programmes with mixed bacterial vaccines in many countries at the present time.     

Religiosity and physical and emotional functioning among African American and White colorectal and lung cancer patients

The literature suggests that religiosity helps cope with illness. The present study examined the role of religiosity in functioning among African Americans and Whites with a cancer diagnosis. Patients were recruited from an existing study and mailed a religiosity survey. Participants (N = 269; 36% African American, 56% women) completed the mail survey, and interview data from the larger cohort was utilized in the analysis. Multivariate analyses indicated that in the overall sample religious behaviors were marginally and positively associated with mental health and negatively with depressive symptoms. Among women, religious behaviors were positively associated with mental health and negatively with depressive symptoms. Religiosity was not a predictor of study outcomes for men. Among African Americans, religious behaviors were positively associated with mental health and vitality. Among Whites, religious behaviors were negatively associated with depressive symptoms. These findings suggest a mixed role of religious involvement in cancer outcomes. The current findings may have applied potential in the areas of emotional functioning and depression.     

VEGF及KDR在大肠腺瘤和腺癌中的表达及意义

目的：探讨VEGF和KDR在大肠腺瘤和大肠腺癌中的表达及临床病理特征的关系。方法：大肠腺瘤和大肠腺癌组织标本各100例,采用免疫组织化学染色法检测VEGF和KDR在标本中的表达情况。结果：VEGF和KDR在大肠腺癌组中的阳性表达明显高于大肠腺瘤组（P〈0.05）;在正常大肠黏膜均未见VEGF和KDR表达的阳性染色;VEGF阳性表达组中KDR的阳性表达率为70%,显著高于VEGF阴性表达组中KDR的阳性表达率16%,两组比较有统计学意义（P〈0.01）。结论：大肠腺癌组织中KDR的表达与肿瘤大小、转移情况、浸润深度密切相关;VEGF和KDR在大肠腺瘤中的表达与患者的年龄、性别及分型均无相关性,而与增生程度相关（P〈0.05）。在大肠腺癌患者中VEGF及KDR表达更高,二者具有协同效应。     

Renal irradiation and the pharmacology and toxicity of methotrexate and cisplatinum

We used a rat model to study the effects of renal irradiation on the pharmacology of methotrexate (MTX) and cisplatinum (cis-Pt). Unanesthetized rats were given bilateral kidney irradiation (20 Gy in 9 fractions). At 9 months after irradiation, 3% of the animals had died and survivors showed moderately impaired renal function. At 15 months, 30% of the animals had died and survivors showed severely impaired renal function. Some animals were given i.v. MTX 1 week to 15 months after irradiation. In irradiated rats, the area under the MTX plasma clearance curve equaled that of controls through 6 months, and was significantly above controls from 9 months on. Other animals were given i.p. cis-Pt 1 week to 9 months after irradiation. The acute toxicity of cis-Pt was the same in control and irradiated rats when cis-Pt was given immediately before or after irradiation. Beginning 3 months after irradiation there was a progressive increase in cis-Pt toxicity and a simultaneous decrease in urinary platinum excretion. Irradiated animals that survived cis-Pt treatment showed increased radiation nephritis; the greatest effect occurred when cis-Pt was given 3 months or more after irradiation. MTX and cis-Pt clearance decreased when renal dysfunction was first observed and changes in renal function preceded changes in drug clearance and toxicity.     

Fruit and vegetables and cancer risk

The possibility that fruit and vegetables may help to reduce the risk of cancer has been studied for over 30 years, but no protective effects have been firmly established. For cancers of the upper gastrointestinal tract, epidemiological studies have generally observed that people with a relatively high intake of fruit and vegetables have a moderately reduced risk, but these observations must be interpreted cautiously because of potential confounding by smoking and alcohol. For lung cancer, recent large prospective analyses with detailed adjustment for smoking have not shown a convincing association between fruit and vegetable intake and reduced risk. For other common cancers, including colorectal, breast and prostate cancer, epidemiological studies suggest little or no association between total fruit and vegetable consumption and risk. It is still possible that there are benefits to be identified: there could be benefits in populations with low average intakes of fruit and vegetables, such that those eating moderate amounts have a lower cancer risk than those eating very low amounts, and there could also be effects of particular nutrients in certain fruits and vegetables, as fruit and vegetables have very varied composition. Nutritional principles indicate that healthy diets should include at least moderate amounts of fruit and vegetables, but the available data suggest that general increases in fruit and vegetable intake would not have much effect on cancer rates, at least in well-nourished populations. Current advice in relation to diet and cancer should include the recommendation to consume adequate amounts of fruit and vegetables, but should put most emphasis on the well-established adverse effects of obesity and high alcohol intakes.     

New and emerging radiosensitizers and radioprotectors

The combination of chemotherapy and radiation has led to clinical breakthroughs in several disease sites, and current work continues to define optimum combinations of proven chemotherapy as well as more recently available, noncytotoxic agents. Administration of systemic therapies allows modulation of radiation response to improve tumor control (radiosensitization) or to prevent normal tissue toxicity (radioprotection). Substantial progress has been made in identifying the targets of standard chemotherapeutic radiation sensitizers and protectors as well as in the introduction of a new generation of molecularly targeted therapies in combination with radiation. We have reviewed the most recent, predominantly early phase clinical trials combining systemic agents with radiation. Although the proof of an improved schedule ultimately needs to come from well-run Phase III trials, the search among schedules could be shortened by the use of surrogate endpoints such as presence of active drug metabolites in the tumor. This has been accomplished only in a few cases and needs to become a more standard part of radiation sensitizer and protector trials.