阿司匹林抑制肺癌细胞增殖的实验研究

 目的 观察阿司匹林在体外对肺腺癌细胞A549增殖的抑制作用。方法 采用噻唑蓝（MTT）法观察阿司匹林及与奥沙利铂联用抑制A549细胞的增殖；采用流式细胞仪（FCM）观察阿司匹林处理后A549细胞周期分布的变化；采用HE染色和DNA末端原位标记染色技术（TUNEL）观察阿司匹林处理后诱导A549细胞凋亡的作用。结果 MTT显示阿司匹林呈剂量依赖方式抑制A549细胞增殖。阿司匹林作用后，FCM显示G0／G1期细胞比例增加，S期和G／M期细胞比例降低，TUNEL显示细胞凋亡指数（AI）由3．67％±1．15％增加到26．33％±2．52％，呈一定剂量效应关系。光镜下可见典型的细胞凋亡形态学变化。阿司匹林（2．5mmol／L）与奥沙利铂（≥6．25ug／mL）联用可增强抑制A549增殖的作用，二者呈协同或相加作用。结论 阿司匹林可抑制肺腺癌细胞A549的增殖，其影响细胞周期分布、诱导细胞凋亡可能是其重要的机制。阿司匹林及与奥沙利铂联用有显著的协同抗增殖效应。     

美加明对人肺腺癌A549细胞增殖和迁移的影响

目的:探讨烟碱型胆碱能受体(nicotinic acetylcholine receptor,nAChR)拮抗剂美加明对人肺腺癌A549细胞增殖和迁移的影响。方法:应用磺酰罗丹明B(sulforhodamine B,SRB)法和FCM法检测美加明对人肺腺癌A549细胞增殖、细胞周期和细胞凋亡的影响;划痕实验检测美加明对细胞迁移能力的影响;激光共聚焦显微镜下观察美加明对A549细胞E-钙黏蛋白(E-cadherin,E-cad)表达的影响。结果:美加明可抑制A549细胞的增殖,与对照组比较差异有统计学意义(P<0.05),并且其抑制作用随着药物浓度的增加而增强。FCM检测结果显示,美加明作用后,G0/G1期细胞比率增多,S期和G2/M期细胞比率下降,而细胞凋亡未发生明显变化。划痕实验结果显示,美加明可降低A549细胞的迁移能力。激光共聚焦显微镜下观察发现,美加明可上调A549细胞E-cad的表达。结论:美加明可能通过阻滞细胞周期于G0/G1期而抑制A549细胞增殖,并通过上调E-cad的表达而降低细胞迁移。     

RNA干扰沉默 NS 基因对人非小细胞肺癌A549细胞增殖和凋亡的影响

目的：研究RNA干扰沉默核干细胞因子（nucleostemin, NS ）基因表达对人肺腺癌A549细胞株增殖和凋亡的影响。方法：向A549细胞内分别转染靶向 NS 基因的siRNA表达载体pcDNA4/C-NS-silencer和空载体pcDNA4/C vector作为silencer组和vector组,以不转染质粒的A549细胞为normal组,Real-time PCR检测转染pcDNA4/C-NS-silencer对A549细胞内 NS 基因表达的影响。CCK-8法检测沉默 NS 基因对A549细胞增殖的影响,流式细胞术检测对细胞周期的影响,Hoechst33258核染色法和流式细胞术检测对细胞凋亡的影响。结果： Silencer组 NS 基因表达较vector组和normal组明显被抑制（0.166±0.024 vs 0.497±0.022、0.505±0.032, 均 P <0.01）;Silencer组A549细胞增殖活性显著低于vector组和normal组 (0.518±0107 vs 0.855±0.102、0.832±0.158,均 P <0.05);Silencer组A549细胞周期阻滞于G0/G1期;Silencer组细胞核皱缩呈致密浓染,染色质碎裂呈块状并有边集现象,且细胞凋亡率较vector组与normal组显著增高\[(34.80±6.77)% vs (9.70±150)%, (8.16±2.01)%, P <0.01\]。 结论： RNA干扰沉默 NS 基因可抑制人肺腺癌A549细胞的增殖,促进细胞凋亡。     

干扰素λ对人肺腺癌细胞A549的增殖抑制作用及其机制研究

目的:探讨干扰素拉姆达(IFNλ)对人肺腺癌细胞A549的增殖抑制作用及其机制,为治疗肺癌提供理论依据.方法:本文以含有IL-1ORl-IFNλR1(10R1-λRl)复合受体的质粒转染人肺腺癌细胞A549.以IL-10刺激转染细胞诱导IFNλ信号表达,在不同时间点进行细胞计数,用流式细胞术分析TUNEL染色、AnnexinV和Pl细胞周期染色,以及活化STAT1的总量.结果:IFNλ对A549细胞增殖的抑制作用优于IFNα和IFNγ.表达10R1-λR1复合受体的A549细胞在IL-10刺激下IFNλ表达增加,TUNEL和AnnexinV染色阳性,PI染色显示细胞周期阻滞在G0/G1期.活化STAT1的表达总量增加,维持活性时间延长.结论:IFNλ信号可诱导人肺腺癌细胞凋亡,可能是通过激活STAT1蛋白实现的.     

基因在肺腺癌A549细胞中的表达及其对细胞生长的抑制

目的: 研究外源性分化抑制因子3（Inhibitor of differentiation 3,Id3）基因表达对人肺腺癌细胞系A549细胞生长的抑制作用。方法：构建Id3和EGFP的融合基因表达载体pEGFP/Id3,采用脂质体转染技术将pEGFP/ Id3导入A549细胞。FCM分析转染细胞EGFP表达效率，荧光显微镜观察EGFP表达情况；半定量RTPCR、免疫细胞化学分析转染后A549细胞 Id3 mRNA和蛋白的表达。MTT法检测转染后A549细胞生长抑制情况，FCM分析A549细胞周期进程，Annexin V/7AAD和Hoechst33258染色检测转染后细胞的凋亡情况。结果:成功构建人Id3与EGFP融合基因表达载体pEGFP/Id3；荧光显微镜和FCM观察到EGFP的有效表达，转染48~72 h时EGFP表达率最高；Id3 mRNA及蛋白在转染后的A549细胞中能有效表达。转染后48 h和72 h，pEGFP/Id3转染组A549细胞生长受到明显抑制（P<0.01）；细胞周期分析表明，pEGFP/Id3转染组G0/G1期细胞显著高于对照组（P<0.05）; Annexin V/7AAD双染结果显示，pEGFP/Id3转染组细胞的早期凋亡率\[（10.67±2.60）%\]显著高于pEGFP组\[（3.39± 2.21）%\]和未转染组\[（2.35 ± 0.95）%\]（P<0.05）；Hoechst33258染色结果也证实pEGFP/Id3组A549细胞出现明显凋亡形态特征。结论:外源性Id3基因在肺腺癌A549细胞中的表达能抑制细胞增殖,并诱导细胞凋亡。     

聚多曲霉菌提取液对培养人肺腺癌细胞增殖作用的实验研究

目的 了解聚多曲霉菌粗提液对人肺腺癌细胞(A549)增殖的影响,并初步探讨其作用机制.方法 采用四甲基偶氮唑蓝法(MTT)检测不同剂量霉菌提取液作用后A549细胞增殖情况.利用琼脂糖凝胶电泳及流式细胞仪(FCM)进行细胞周期分析,并检测细胞凋亡率.结果 MTT法结果显示,大剂量霉菌提取液可显著抑制A549细胞的增殖,并呈剂量依赖效应;小剂量可促进细胞增殖.FCM检测结果显示,大剂量干预组G0/G1期细胞显著增加,S期细胞显著减少,并在G1期前出现细胞凋亡图像;小剂量干预组G0/G1期细胞显著减少,S期细胞显著增加;G2期细胞均无明显改变.结论 聚多曲霉菌提取液对A549细胞的增殖呈双向作用;除诱导细胞凋亡外,聚多曲霉菌产物的细胞毒性或其他生物学效应可能是其发挥抑制A549细胞增殖作用的更主要途径.     

丹参酮ⅡA对MKN-45细胞生长的影响

目的:研究丹参酮ⅡA(Tanshi-noneⅡA,TanⅡA)对人胃癌细胞株MKN-45的生长抑制作用。方法:选用人胃癌细胞株MKN-45,运用MTT法、流式细胞术(FCM)和逆转录聚合酶链式反应(RT-PCR)半定量法检测细胞增殖、细胞周期、细胞凋亡和各组p53mRNA表达。结果:TanⅡA可明显抑制MKN-45细胞的增殖,其抑制作用具有时间-效应和剂量-效应关系。当2·0μg/mL TanⅡA处理MKN-45细胞96h,增殖抑制率达(74·84±0·41)%。FCM检测肿瘤细胞DNA变化,观察到TanⅡA使MKN-45细胞周期阻滞于G2/M期,出现浓度依赖性的亚“G1”峰,同时野生型p53表达增加。结论:TanⅡA能有效地抑制人胃癌细胞株MKN-45的增殖,其可能的机制与凋亡有关。肿瘤防治杂志,2005,12(19):1465-1468     

STI 571增强非小细胞肺癌对顺铂的敏感性

目的:比较STI 571单药及联合顺铂(DDP)对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞株A549及其DDP耐药株(A549/DDP)的增殖、细胞周期和凋亡的影响以及STI 571相关受体在NSCLC组织中的表达.方法:体外培养A549和A549/DDP细胞,应用MTT法测定STI 571和(或)DDP对细胞的增殖作用;通过FCM法分析细胞周期和凋亡;应用免疫细胞化学和免疫组织化学染色分别测定A549细胞株和NSCLC组织中STI 571相关受体的表达.根据Kaplan-Meier生存曲线,比较血小板衍化生长因子受体(platelet-derived growth factor receptor,PDGFR)-α和-β的表达与NSCLC患者总体生存的关系.结果:STI 571单药对A549/DDP细胞有增殖抑制作用,能增强细胞对DDP的敏感性,增加A549/DDP细胞的G2/M期和S期的细胞数,诱导其凋亡.STI 571在<10 μmol/L时,对A549细胞增殖无明显影响.A549细胞和A549/DDP细胞均表达PDGFR-α和-β,后者还表达c-KIT蛋白.在肺腺癌中,PDGFR-α和PDGFR-β的表达率分别为65.5%和69.0%,与肺鳞癌的70.4%和74.1%相似,仅1例肺鳞癌表达c-KIT(3.7%).表达PDGFR-α和-β的患者3年生存率和总体生存与不表达患者相似.结论:STI 571可以抑制DDP耐药的NSCLC细胞株的增殖,诱导其凋亡,并能增加DDP耐药细胞对DDP的敏感性.在NSCLC患者中,PDGFR-α和-β的表达水平与预后无关.     

聚多曲霉菌提取液对培养人肺腺癌细胞增殖作用的实验研究

目的：了解聚多曲霉菌粗提液对人肺腺癌细胞（A549）增殖的影响,并初步探讨其作用机制。方法：采用四甲基偶氮唑蓝法（MTT）检测不同剂量霉菌提取液作用后A549细胞增殖情况。利用琼脂糖凝胶电泳及流式细胞仪（FCM）进行细胞周期分析,并检测细胞凋亡率。结果：MTT法结果显示,大剂量霉菌提取液可显著抑制A549细胞的增殖,并呈剂量依赖效应;小剂量可促进细胞增殖。FCM检测结果显示,大剂量干预组G0/G1期细胞显著增加,S期细胞显著减少,并在G1期前出现细胞凋亡图像;小剂量干预组G0/G1期细胞显著减少,S期细胞显著增加;G2期细胞均无明显改变。结论：聚多曲霉菌提取液对A549细胞的增殖呈双向作用;除诱导细胞凋亡外,聚多曲霉菌产物的细胞毒性或其他生物学效应可能是其发挥抑制A549细胞增殖作用的更主要途径。     

白藜芦醇对肺腺癌A549细胞增殖、黏附与侵袭的影响

目的:研究白藜芦醇(resveratrol,Res)对肺腺癌A549细胞增殖,黏附及侵袭的影响。方法:用不同剂量的Res作用于A549细胞,MTT法测定A549细胞的增殖水平(成纤维3T3细胞为对照),流式细胞仪检测A549细胞的细胞周期和凋亡,体外黏附实验测定A549细胞的黏附能力,Transwell实验测定A549细胞的侵袭能力,荧光免疫细胞化学方法测定A549细胞中MMP-2和TIMP-2蛋白的表达。结果:Res以剂量(20～80μmol/L)依赖和时间(0～72 h)依赖方式抑制A549细胞的增殖,同样条件对3T3细胞增殖无影响。10、20、40、80μmol/L Res作用后,A549细胞的凋亡率分别为(34.9±0.91)%、(41.33±0.13)%、(45.47±0.87)%和(59.46±0.59)%。经20μmol/L Res处理48 h后,S期A549细胞比例为(56.41±1.67)%,细胞周期阻滞在S期。20μmol/L以上的Res可引起A549细胞的黏附力下降、侵袭力降低(P<0.05);同时,A549细胞内MMP-2蛋白表达下调,而TIMP-2蛋白表达增加。结论:Res抑制肺腺癌A549细胞的增殖、黏附和侵袭,其机制可能涉及对MMP-2和TIMP-2表达的双向调控。     

Aspirin and lung cancer in women

The association between aspirin use and lung cancer risk in women was examined in a case-control study nested in the New York University Women's Health Study, a large cohort in New York. Case subjects were all the 81 incident lung cancer cases who had provided information about aspirin use at enrollment and during the 1994-1996 follow up. Ten controls per case were randomly selected from among study participants who matched a case by age, menopausal status, and dates of enrollment and follow-up. Relative to no aspirin use, the odds ratio for lung cancer (all histological sub-types combined) among subjects who reported aspirin use three or more times per week for at least 6 months was 0.66 (95% confidence interval 0.34-1.28), after adjustment for smoking and education. A stronger inverse association was observed in analyses restricted to non-small cell lung cancer (adjusted odds ratio 0.39, 95% confidence interval 0.16-0.96). These results suggest that regular aspirin use might be inversely associated with risk of lung cancer in women, particularly the non-small cell sub-type.     

Aspirin use and cancers of the upper aerodigestive tract

The role of aspirin on the risk of cancers of the upper aerodigestive tract was investigated in the combined data of three Italian case-control studies, including 965 cases and 1779 hospital controls. The odds ratio was 0.33 for users of > or = 5 years, and 0.51 for > or = 5 years since first use.     

Hydrogen sulfide-releasing aspirin inhibits the growth of leukemic Jurkat cells and modulates β-catenin expression

Hydrogen sulfide-releasing aspirin (HS-ASA) is a novel compound with potential against cancer. It inhibited the growth of Jurkat T-leukemia cells with an IC₅₀ of 1.9 ± 0.2 μM whereas that of ASA was >5000 μM. It dose-dependently inhibited proliferation and induced apoptosis in these cells, causing a G₀/G₁ cell cycle arrest. HS-ASA down-regulated β-catenin protein levels and reduced mRNA and protein expression of β-catenin/TCF downstream target genes cyclinD1 and c-myc. Aspirin up to 5 mM had no effect on β-catenin expression. HS-ASA also increased caspase-3 protein levels and dose-dependently increased its activity. These effects were substantially blocked by z-VAD-fmk, a pan-caspase inhibitor.     

Aspirin and risk for gastric cancer: a population-based case-control study in Sweden

While aspirin and other non-steroid anti-inflammatory drugs (NSAIDs) are associated with gastric mucosal damage, they might reduce the risk for gastric cancer. In a population-based case-control study in 5 Swedish counties, we interviewed 567 incident cases of gastric cancer and 1165 controls about their use of pain relievers. The cases were uniformly classified to subsite (cardia/non-cardia) and histological type and information collected on other known risk factors for gastric cancer. Helicobacter pylori serology was tested in a subset of 542 individuals. Users of aspirin had a moderately reduced risk of gastric cancer compared to never users; odds ratio (OR) adjusted for age, gender and socioeconomic status was 0.7 (95% CI = 0.6-1.0). Gastric cancer risk fell with increasing frequency of aspirin use (P for trend = 0.02). The risk reduction was apparent for both cardia and non-cardia tumours but was uncertain for the diffuse histologic type. No clear association was observed between gastric cancer risk and non-aspirin NSAIDs or other studied pain relievers. Our finding lends support to the hypothesis that use of aspirin reduces the risk for gastric cancer.     

Relationship between aspirin use after diagnosis of colorectal cancer and patient survival: a meta-analysis of observational studies

Background:

Epidemiological evidence suggests that use of aspirin after the diagnosis of colorectal cancer can lengthen survival. However, the supporting data vary between studies, and this hypothesis remains controversial. We conducted a meta-analysis to provide a quantitative assessment of the association between use of aspirin after diagnosis of colorectal cancer and patient survival.

Methods:

We searched the Medline and Embase databases up to April 2014 to identify studies related to aspirin use after diagnosis and all-cause mortality or colorectal cancer-specific mortality. Summary effect estimates with 95% confidence intervals (CIs) were derived using a fixed or random effects model, depending on the heterogeneity between the included studies.

Results:

Seven epidemiologic studies that consisted of six cohort studies and one nested case–control study were included in this meta-analysis. The hazard ratio (HR) of the association between aspirin use after colorectal cancer diagnosis and overall mortality, which was reported in five studies, was 0.74 (95% CI, 0.62–0.89) using a random model (heterogeneity test P=0.003, I²=75.3%), and for colorectal cancer-specific mortality (four studies), it was 0.75 (95% CI, 0.51–1.10) using a random model (heterogeneity test P=0.001, I²=84.1%). In addition, we analysed postdiagnosis aspirin use according to whether aspirin was also used before diagnosis. The HR for the overall mortality of patients who did not use aspirin before diagnosis, which was reported in four studies, was 0.84 (95% CI, 0.70–1.00), and for colorectal cancer-specific mortality (three studies), it was 0.79 (95% CI, 0.61–1.02). For those who did use aspirin before diagnosis, the HR for overall mortality (four studies) was 0.88 (95% CI, 0.83–0.93), and for colorectal cancer-specific mortality (three studies), it was 0.80 (95% CI, 0.59–1.09). Subgroup analysis showed that use of aspirin after diagnosis was associated with longer overall survival among patients with the variant PIK3CA gene but not for those with wild-type PIK3CA.

Conclusions:

Based on current evidence, the use of aspirin after diagnosis does not reduce colorectal cancer-specific mortality, but it does reduce all-cause mortality for colorectal cancer patients.     

Cyclooxygenase inhibitors use is associated with reduced risk of esophageal adenocarcinoma in patients with Barrett's esophagus: a meta-analysis

Background:

Esophageal adenocarcinoma (EAC) has high mortality and is increasing in incidence. Barrett''s esophagus (BE) increases the risk for EAC. Studies have reported inconsistent findings on the association between use of cyclooxygenase (COX) inhibitors and the risk of neoplastic progression in BE patients. Therefore, we performed a meta-analysis to investigate this association.

Methods:

A meta-analysis was undertaken among a total of 9 observational studies using fixed- and random-effects models, comprising 5446 participants; 605 had EAC or high-grade dysplasia (HGD).

Results:

Overall, COX inhibitors use was associated with a reduced risk of EAC/HGD among BE patients (relative risk (RR)=0.64, 95% confidence interval (CI)=0.53–0.77). Aspirin use also reduced the risk of EAC/HGD (RR=0.63, 95% CI=0.43–0.94), as well as non-aspirin COX inhibitors (RR=0.50, 95% CI=0.32–0.78). The chemopreventive effect seemed to be independent of duration response.

Conclusions:

Cyclooxygenase inhibitors use is associated with a reduced risk of developing EAC in patients with BE. Both low-dose aspirin and non-aspirin COX inhibitors are associated with a reduced risk of neoplasia. More well-designed randomised controlled trials are needed to increase our understanding of the chemopreventive effect of COX inhibitors.     

阿司匹林对胃癌细胞株SGC-7901作用的相关机理研究

目的 :探讨阿司匹林 (ASA)对胃癌细胞株SGC 790 1作用的相关机理 ,为寻找其新的药物用途奠定理论基础。方法 :用MTT法检测药物对细胞的抑制率 ;用流式细胞仪测定细胞周期 ;用琼脂糖凝胶电泳法检测细胞凋亡 ;3H TdR同位素示踪测定细胞DNA合成。结果 :ASA对SGC 790 1细胞株的细胞作用具有量效和时效依赖关系。ASA对细胞DNA合成有抑制作用 ,并可使SGC 790 1细胞周期中S期及G2 /M期比例升高 ,G1期比例下降 ,呈一定的剂量效应关系。琼脂糖凝胶电泳结果未见典型的阶梯状凋亡条带。结论 :阿司匹林对SGC 790 1细胞株存在着细胞毒作用 ,其细胞毒作用可能与抑制DNA的合成、阻止细胞周期有关系     

阿司匹林、扑热息痛和尼美舒利对人卵巢癌细胞株SKOV3的抑制作用

[目的]研究非甾体类药物阿司匹林、扑热息痛和尼美舒利对人卵巢癌细胞株SKOV3的抑制作用及其相关机制.[方法]利用四甲基偶氮唑蓝(MTT)法观察阿司匹林、扑热息痛和尼美舒利对人卵巢癌细胞株SKOV3的生长抑制作用;免疫组化法检测COX-2在SKOV3细胞中的表达情况以及3种药物对SKOV3细胞中Ki-67,C-erbB-2表达的影响;透射电镜观察3种药物作用SKOV3细胞后形态变化和凋亡情况;流式细胞术检测3种药物对SKOV3细胞周期的影响.[结果]阿司匹林、扑热息痛和尼美舒利对人卵巢癌细胞株SKOV3具有生长抑制作用,并呈时间-剂量依赖性.免疫组化发现COX-2在SKOV3细胞中呈阳性表达,阿司匹林使SKOV3细胞中C-erbB-2蛋白表达下降,尼美舒利使SKOV3细胞核中Ki-67蛋白表达下降,扑热息痛对C-erbB-2和Ki-67蛋白表达无影响.透射电镜可见阿司匹林、扑热息痛和尼美舒利作用后的SKOV3细胞中有凋亡小体和坏死细胞的形成.流式细胞术发现400μmol/L尼美舒利和10-2mol/L阿司匹林使SKOV3细胞停滞在G0/G1期.[结论]非甾体类药物,特别是COX-2抑制剂尼美舒利可望作为预防性化疗药物应用于卵巢癌的治疗.     

阿司匹林对结肠癌细胞凋亡及Fas/FasL表达的影响

目的：探讨阿司匹林对结直肠癌化学预防的机理。方法：培养结肠癌细胞株Caco-2，用不同浓度阿司匹林进行干预，对细胞生长、增殖、凋亡及Fas／FasL表达进行观察、检测。结果：高浓度阿司匹林组细胞大量死亡。低浓度组促进细胞凋亡；25mmol／L阿司匹林组可见Fas表达，而对照组不表达；25mmol／L阿司匹林组FasL表达与对照组相比明显降低。结论：低浓度阿司匹林促进结肠癌细胞凋亡，其机理与诱导Fas表达、下调FasL表达有关。     

大剂量甲氨蝶呤治疗成人急性淋巴细胞白血病的临床观察

探讨使用大剂量甲氨蝶呤(HD -MTX)在成人急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)维持强化治疗过程中的疗效和毒副反应,采用甲氨蝶呤(MTX)1~3 g/m2,24 h持续静脉滴入,用来成人ALL的强化治疗,并用四氢叶酸钙(CF)解救。结果呈缓解状态86 .0%(49/57),骨髓复发7 0%(4/57),中枢神经系统白血病(CNS L)1 .8%(1/57),死亡5. 2%(3/57)。初步研究结果提示,HD- MTX治疗成人ALL疗效肯定,相对骨髓抑制较轻,黏膜皮肤损害较突出,毒副反应可以耐受。