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Effect of optimal combination of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) on proliferation and apoptosis of a549 lung cancer cells

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Journal of Traditional Chinese Medicine 2018年第3期38卷 351-358页

作者： Xu Chengyong Wang Yuguo Feng Jian Qin Li Xu Ran Dou Yongqi Medical School of Chinese People's Liberation Army Chinese People's Liberation Army General Hospital Beijing 100853 China Department of Traditional Chinese Medicine Hainan Branch of Chinese People's Liberation Army General Hospital Sanya 572013 China Department of Traditional Chinese Medicine Chinese People's Liberation Army General Hospital Beijing100853 China

OBJECTIVE: To investigate the effect of optimal combination(E) of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) on proliferation and apoptosis of a549 lung cancer cells and the possible mechanism underpinning the ***: A uniform design method was used to optimize the E of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) in a549 lung cancer cells. MTS assay was applied to analyze the effect of the component formula ofHuangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) on a549 cells viability in various uniform design groups. a549 cells with exponential growth in routine culture were exposed to Co Cl_2(200 μmol/L) to mimic hypoxic conditions. Group 0 was treated with RPMI-1640, the group Co Cl_2 was treated with Co Cl_2(200 μmol/L),the group DDP + Co Cl_2 was treated with 4 mg/L Cisplatin injection(DDP) + Co Cl_2(200 μmol/L), and the drug group was treated with various dose of E(0.5 E, 1 E, 2 E) + Co Cl_2(200 μmol/L). All groups were cultured for 24 h. Cell apoptosis was measured by Annexin V-FITC/propidium iodide double staining and flow cytometry. Western blot assay and quantitative real-time polymerase chain reaction(q RT-PCR) were employed to detect the protein and m RNA expression of B-celllymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax) and cysteinyl aspartate specific proteinase-3(caspase-3).RESULTS: The E obtained by the uniform design was comprise of 200 mg/L Astragalus polysaccharide(X1) and 32 mg/L Curcumin(X3). Group DDP+Co Cl_2, group 1 E + Co Cl_2 and group 2 E + Co Cl_2 promoted the apoptosis of a549 cells(P 0.05). Compared with group 0, various doses of E + Co Cl_2 could up-regulate the expression of Bax and caspase-3 and down-regulate the expression of Bcl-2 at protein and m RNA levels(P < 0.05).CONCLUSION: Astragalus polysacchari

关键词： Lung neoplasms a549 cells Apoptosis Astragalan Huangqi (Radix Astragali Mongolici) Ezhu (Rhizoma Curcumae Phaeocaulis) Curcumin Hypoxia

Gene Expression Profiles Comparison between 2009 Pandemic and Seasonal H1N1 Influenza Viruses in a549 Cells

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Biomedical and Environmental Sciences 2010年第4期23卷 259-266页

作者： XIAO-XING YANG NING DU JIAN-FANG ZHOU ZI LI MIN WANG JUN-FENG GUO DA-YAN WANG YUE-LONG SHU State Key Laboratory for Molecular Virology and Genetic Engineering National Institute for Viral Disease Control and Prevention China CDC 100 Yingxin Street Xuanwu District Beijing 100052 China

Objective To perform gene expression profiles comparison so that to identify and understand the potential differences in pathogenesis between the pandemic and seasonal A （H1N1） influenza viruses. Methods a549 cells were infected with A/California/07/09 （H1N1） and A/GuangdongBaoan/51/08 （H1N1） respectively at the same MOI of 2 and collected at 2, 4, 8, and 24 h post infection （p.i.）. Gene expression profiles of a549 cells were obtained using the 22 K Human Genome Oligo Array, and differentially expressed genes were analyzed at selected time points. Results Microarrays results indicated that both of the viruses suppressed host immune response related pathways including cytokine production while pandemic H1N1 virus displayed weaker suppression of host immune response than seasonal H1N1 virus. Observation on similar anti-apoptotic events such as activation of apoptosis inhibitor and down-regulation of key genes of apoptosis pathways in both infections showed that activities of promoting apoptosis were different in later stage of infection. Conclusion The immuno-suppression and anti-apoptosis events of pandemic H1N1 virus were similar to those seen by seasonal H1N1 virus. The pandemic H1N1 virus had an ability to inhibit biological pathways associated with cytokine responses, NK activation and macrophage recognition .

关键词： Influenza A virus Pandemic H1N1 a549 Expression profiling array

Inhibitory Effects of Natural Compound Alantolactone on Human Non-small Cell Lung Cancer a549 Cells

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Chemical Research in Chinese Universities 2011年第2期27卷 241-244页

作者： ZONG Min-ru ZHAO Ying-hao ZHANG Kun YANG Long-fei ZHENG Yong-chen HE Cheng-yan China-Japan Union Hospital of Jilin University Changchun 130033 P. R. China The Second Hospital of Jilin University Changchun 130041 P. R. China

Alantolactone is a natural compound identified from the roots of Inula helenium L. that has multiple bio-activities. We examined its inhibitory effects on human non-small cell lung cancer（NSCLC） a549 cells. The an-tiproliferative effect of alantolactone on a549 cells was investigated via MTT[3′-（4,5dimethylthiazol-2-yl）-2,5- diphenyl tetrazolium bromide] assay and its apoptosis-inducing effect was determined by Hoechst staining and flow cytometry. We found that alantolactone significantly inhibited the proliferation of a549 cells and induced morphological changes typical for apoptosis. Flow cytometry analysis indicates dose-dependent cell cycle retardation at G0/G1 and S stages. The results indicate that alantolactone could be an attractive small-molecular natural compound for further development as a therapeutic drug against NSCLC.

关键词： Alantolactone Non-small cell lung cancer(NSCLC) a549 Proliferation Apoptosis

Epithelial mesenchymal transition of non-small-cell lung cancer cells a549 induced by SPHK1

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Asian Pacific Journal of Tropical Medicine 2015年第2期8卷 142-146页

作者： Min Ni Xiao-Lei Shi Zhi-Gang Qu Hong Jiang Zi-Qian Chen Jun Hu Third Department of Health Care for Cadres Fuzhou General Hospital of Nanjing Military Region Second Hospital Affiliated to University of South China

Objective:To explore the effect and molecular mechanism of SPHK1 in the invasion and metastasis process of non-small-cell lung cancer cells(a549).Methods:Recombinant retrovirus was used to mediate the production of a549/vector,a549/SPHK1,a549/scramble,and a549/SPHK1/RNAi that stably expressed or silenced *** invasion and migration capacities of a549 cells overexpressing or silencing SPHK1 were determined using Transwell invasion assay and scratch wound repair *** protein and mRNA expression levels of E-cadherin,fibronectin,vimentin in a549/vector,a549/SPHK1,a549/scramble,a549/SPHK1/RNAi were detected with Western blot(WB) and quantitative PCR(QPCR) methods,***:Transwell invasion assay and scratch wound repair experiments showed that over-expression of SPHK1 obviously enhanced the invasion and migration capacities of a549 *** and QPCR detection results showed that,the expression of E-cadherin(a molecular marker of epithelial cells) and fibronectin,vimentin(molecular markers of mesenchymal cells) in a549 cells was upregulated after overexpression of SPHK1;while SPHK1 silencing significantly reduced the invasion and metastasis capacities of a549 cells,upregulated the expression of molecular marker of epithelial cells,and downregulated the expression of molecular marker of mesenchymal ***:SPHK1 promotes epithelial mesenchymal transition of non-small-cell lung cancer cells and affects the invasion and metastasis capacities of these cells.

关键词： SPHK1 Epithelial mesenchymal transition a549 Invasion Metastasis

Effect of 5-AZn-2'-deoxycytidine on proliferation of human lung adenocarcinoma cell line a549 in vitro

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Asian Pacific Journal of Tropical Medicine 2013年第12期6卷 982-985页

作者： Na Li Hua-Qiong Huang Gen-Sheng Zhang Wei Cui Department of Intensive Care Unit Second Affiliated Hospital of Zhejiang University School of Medicine Department of Respiratory Medicine Second Affiliated Hospital of Zhejiang Unirersity School of Medicine

Objective:To explore effect of 5-AZn-2'-deoxycytidine on proliferation of human lung adenocarcinoma cell line a549 in ***:Superoxide dismutase(SOD)activity was measured by hydroxylamine colorimetric *** effect of 5-AZn-2'deoxycytidylic acid at different concentration and different time on growth of a549 cell was determined by MTT *** dioxyamphetamine(MDA)was measured by throbarbituric acid colorimetric *** of 5-AZn-2'deoxycytidylic acid on apoptosis of a549 cell was determined by Hoechst 33258 dyeing ***:5-AZn-2'deoxycytidylic acid had significant inhibition effect on proliferation of a549 cells in vitro,and the inhibition was notably dependent on time and dosage during 48-72 h;SOD level was significantly lower than those of control group(Pa549 cells began to be in apoptosis after using 5-AZn-2'deoxycytidylic ***:5-AZn-2'deoxycytidylic acid has significant inhibition effect on growth of a549 cell,and can lead the change of lipid *** indicates that the mechanism has relationship with a549 cell cycle tissue and induction factor of apoptosis.

关键词： DAC a549 cell Lipid peroxidation Apoptosis

Differential alterations of positive and negative regulators of beta catenin enhance endogenous expression and activity of beta catenin in a549 non small cell lung cancer(NSCLC)cells

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Genes & Diseases 2016年第4期3卷 282-288页

作者： Supratim Ghatak Sanghamitra Raha Crystallography and Molecular Biology Division Saha Institute of Nuclear Physics1/AF BidhannagarKolkata 700064India Department of Biotechnology and ISERC Visva Bharati UniversitySantiniketan731235India Present address:Centre for Cellular and Molecular Biology Uppal RoadHyderabad 500007India

Beta catenin has been well documented in previous studies to be involved in non small cell lung cancer(NSCLC).Beta catenin abundance and transcriptional activity are significantly regulated by several *** it is well known that Akt and Gsk3 beta are respective positive and negative regulators of beta catenin,however,no single study has so far documented how the expression and activity of both positive as well as negative regulators play favorable role on beta catenin expression and activity in *** this study,we compared expression and activity of beta catenin and its regulators in normal lung cell WI38 and NSCLC cell a549 by western blot,qRT-PCR and luciferase *** observed that beta catenin positive regulators(Akt and Hsp90)and negative regulators(Gsk3 beta and microRNA-214)have differential expression and/or activity in NSCLC cell *** the differentially altered statuses of both the positive and negative regulators rendered cumulative positive effect on beta catenin expression and activity in *** study thus suggests that chemotherapeutic modulations of regulating factors are crucial when abrogation and/or inhibition of key oncogenic proteins are necessary for cancer chemotherapy.

关键词： a549 Beta catenin Gsk3 beta miR-214 Non small cell lung cancer

北京大气PM_(2.5)对a549细胞炎性因子及DNA损伤的毒性

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中国环境科学 2016年第5期36卷 1579-1588页

作者：焦周光付绪磊温占波李劲松李娜张柯王洁胡凌飞军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室北京100071 渤海大学食品科学与工程学院辽宁锦州121013

为探讨大气PM_(2.5)及其不同组分对人肺上皮细胞a549的毒性作用及其剂量-反应关系,将前期采集的PM_(2.5)颗粒物用不同方法进一步制备PM_(2.5)水溶性组分、PM_(2.5)脂溶性组分和PM_(2.5)单纯颗粒物,将制备的PM_(2.5)颗粒物及其组分以不... 详细信息

为探讨大气PM_(2.5)及其不同组分对人肺上皮细胞a549的毒性作用及其剂量-反应关系,将前期采集的PM_(2.5)颗粒物用不同方法进一步制备PM_(2.5)水溶性组分、PM_(2.5)脂溶性组分和PM_(2.5)单纯颗粒物,将制备的PM_(2.5)颗粒物及其组分以不同浓度(10,50,100,200,400μg/m L)对a549细胞染毒,用MTS法分别在染毒6,10,24,48,72h后测定细胞活力,染毒24h后用ELISA及RT-QPCR法测定炎性因子IL-6和TNF-α表达量,AP位点计数法测定细胞DNA损伤情况.结果表明:除PM_(2.5)水溶性组分外,其余染毒样本高浓度染毒时始终对细胞生长表现出抑制作用,其中低浓度染毒时可在较短时间对细胞生长表现出抑制作用,染毒时间较长时抑制作用减弱或消失,PM_(2.5)水溶性组分对细胞生长抑制作用并不显著;除PM_(2.5)水溶性组分外,其余染毒样本都显著升高了IL-6m RNA的相对表达量和IL-6蛋白的分泌,除PM_(2.5)脂溶性组分外,其余染毒样本都显著升高了TNF-αm RNA的相对表达量;除PM_(2.5)水溶性组分外,其余染毒样本都显著提高了DNA碱基缺失程度.总的来说,PM_(2.5)水溶性组分在抑制细胞活力、造成炎性损伤及DNA损伤方面作用相对较小,而PM_(2.5)所产生的毒性作用并不仅限于其所吸附的复杂成分,其中作为载体的固体核心颗粒对机体可能造成的毒性作用也不容忽视.

关键词： PM2.5 组分 a549 细胞活力炎性因子 DNA损伤

榄香烯乳逆转肺腺癌a549/DDP细胞耐药性及可能作用机制的研究

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肿瘤 2013年第12期33卷 1061-1068页

作者：姚成才涂远荣杜好信姜杰叶社房张义陈隽鹏福建中医药大学附属厦门中医院胸外科福建厦门361009 福建医科大学附属第一医院胸外科福建福州355000 厦门大学附属第一医院胸外科福建厦门361001 厦门大学分子生物学与材料学院福建厦门361000

目的 :探讨榄香烯乳(elemene)逆转人肺腺癌耐顺铂(cisplatin,DDP)细胞a549/DDP的耐药性及可能的作用机制。方法 :采用MTT法检测榄香烯乳单药作用时的细胞毒作用及与DDP联合作用对a549/DDP细胞耐药的逆转作用;Hoechst 33342法进行细胞核... 详细信息

目的 :探讨榄香烯乳(elemene)逆转人肺腺癌耐顺铂(cisplatin,DDP)细胞a549/DDP的耐药性及可能的作用机制。方法 :采用MTT法检测榄香烯乳单药作用时的细胞毒作用及与DDP联合作用对a549/DDP细胞耐药的逆转作用;Hoechst 33342法进行细胞核染色,荧光显微镜下观察细胞形态的改变;FCM检测细胞凋亡率;蛋白质印迹法检测细胞质中细胞色素C(cytochrome C,cyt C)、pro-caspase-3、caspase-3和抗凋亡因子B细胞淋巴瘤-2(B cell lymphoma-2,Bcl-2)家族蛋白表达的情况。结果 :不同浓度榄香烯乳对a549/DDP细胞均有一定的抑制作用,呈时间-剂量依赖性效应,联合DDP能提高耐药细胞株a549/DDP对DDP的敏感性而逆转其耐药性。Hoechst 33342法和FCM法检测结果显示,榄香烯乳联合DDP能更大程度上诱导a549/DDP细胞的凋亡,提高耐药细胞的凋亡率,同时还上调a549/DDP细胞质中cyt C、caspase-3和Bad蛋白的表达,而下调pro-caspase-3和Bcl-2蛋白的表达。结论 :榄香烯乳能逆转a549/DDP细胞耐药性可能与其损伤线粒体膜,使线粒体释放cyt C到细胞质、同时活化caspase-3、上调促凋亡蛋白Bad和下调抗凋亡蛋白Bcl-2表达以启动凋亡路径有关。

关键词：肺肿瘤抗药性肿瘤细胞凋亡榄香烯 a549 DDP细胞

丙泊酚下调水通道蛋白3和基质金属蛋白酶-9表达抑制人肺癌a549细胞的侵袭力

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南方医科大学学报 2016年第9期36卷 1286-1290,1294页

作者：叶慧瑾白建杰郭培培汪威林春水南方医科大学南方医院麻醉科广东广州510515

目的探讨丙泊酚对人肺癌a549细胞水通道蛋白3(AQP-3)、基质金属蛋白酶-9(MMP-9)表达及其侵袭力的影响。方法a549细胞经丙泊酚不同剂量(25、50、100μmol/L)和不同时间(12、24 h)处理。用RT-PCR方法观察丙泊酚对a549细胞AQP-3m RNA的作用... 详细信息

目的探讨丙泊酚对人肺癌a549细胞水通道蛋白3(AQP-3)、基质金属蛋白酶-9(MMP-9)表达及其侵袭力的影响。方法a549细胞经丙泊酚不同剂量(25、50、100μmol/L)和不同时间(12、24 h)处理。用RT-PCR方法观察丙泊酚对a549细胞AQP-3m RNA的作用,用Western Blot方法和Transwell侵袭实验分别检测不同浓度丙泊酚作用24 h对a549细胞AQP-3、MMP-9蛋白表达和细胞侵袭力的影响。结果与空白组比较,25、50、100μmol/L丙泊酚24 h处理组对AQP-3 m RNA表达的最大抑制比值为0.13,最小0.41;12 h处理组最大抑制比值0.19,最小0.65,差异有统计学意义(Pa549细胞AQP-3 m RNA、AQP-3蛋白和MMP-9蛋白的表达,抑制a549细胞的侵袭力。

关键词：丙泊酚水通道蛋白3 基质金属蛋白酶9 a549 侵袭力