[1]刘　冲,郜赵伟,王会平,等.腺苷对肺癌细胞增殖的影响及其机制[J].新乡医学院学报,2020,37(8):701-706.[doi:10.7683/xxyxyxb.2020.08.001]
　LIU Chong,GAO Zhaowei,WANG Huiping,et al.Effect of adenosine on proliferation of lung cancer cells and its mechanism[J].Journal of Xinxiang Medical University,2020,37(8):701-706.[doi:10.7683/xxyxyxb.2020.08.001]
点击复制

腺苷对肺癌细胞增殖的影响及其机制
分享到：

《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
37
期数:
2020年8
页码:
701-706
栏目:
基础研究
出版日期:
2020-08-05

文章信息/Info

Title:
Effect of adenosine on proliferation of lung cancer cells and its mechanism
作者:
刘　冲; 郜赵伟; 王会平; 何　婷; 刘　丽; 马海航; 丁聪聪; 董　轲
（空军军医大学第二附属医院检验科，陕西　西安　710038）
Author(s):
LIU Chong; GAO Zhaowei; WANG Huiping; HE Ting; LIU Li; MA Haihang; DING Congcong; DONG Ke
(Department of Clinical Laboratory,the Second Affiliated Hospital of Air Force Medical University,Xi′an 710038,Shannxi Province,China)
关键词:
腺苷; 增殖; 周期; 迁移; 凋亡
Keywords:
adenosine; proliferation; cycle; migration; apoptosis
分类号:
R734.2
DOI:
10.7683/xxyxyxb.2020.08.001
文献标志码:
A
摘要:
目的　研究腺苷对肺癌细胞增殖的作用，并探究其潜在分子机制。方法　取对数生长期A549细胞分为空白对照组和0.5、1.0、2.5、5.0 mmol·L-1腺苷处理组，分别加入含0.0、0.5、1.0、2.5、5.0 mmol·L-1腺苷的培养液进行培养。采用细胞计数试剂盒-8检测腺苷对细胞增殖的影响，流式细胞术检测腺苷对A549细胞周期和凋亡的影响，划痕修复实验检测细胞迁移能力，实时荧光定量聚合酶链反应检测腺苷对A549细胞凋亡相关分子表达水平的影响。结果　0.5、1.0、2.5、5.0 mmol·L-1腺苷处理组A549细胞培养24、48 h的生长抑制率均显著高于空白对照组（P<0.05）；各浓度腺苷处理组A549细胞培养24、48 h的生长抑制率随浓度的升高逐渐增加，两两比较差异均有统计学意义（P<0.05）。2.5、5.0 mmol·L-1腺苷处理组A549细胞相对迁移率均显著低于空白对照组(P<0.05)；5.0 mmol·L-1腺苷处理组A549细胞相对迁移率显著低于2.5 mmol·L-1腺苷处理组(P<0.05)。与空白对照组比较，2.5 mmol·L-1腺苷处理组A549细胞处于G0/G1期比例显著降低，S期比例升高，G2/M期比例升高 (P<0.05)；5.0 mmol·L-1腺苷处理组A549细胞处于G0/G1期比例显著降低，G2/M期比例显著升高 (P<0.05) ；空白对照组和5.0 mmol·L-1腺苷处理组A549细胞处于S期比例比较差异无统计学意义 (P>0.05)。与2.5 mmol·L-1腺苷处理组比较，5.0 mmol·L-1腺苷处理组A549细胞处于S期比例降低，G2/M期比例升高 (P<0.05)；2.5 mmol·L-1腺苷处理组和5.0 mmol·L-1腺苷处理组细胞处于G0/G1期比例比较差异无统计学意义 (P>0.05)。2.5、5.0 mmol·L-1腺苷处理组A549细胞凋亡率显著高于空白对照组 (P<0.05)；5.0 mmol·L-1腺苷处理组A549细胞凋亡率显著高于2.5 mmol·L-1腺苷处理组 (P<0.05)。5.0 mmol·L-1腺苷处理组细胞中Bak、Caspase-3 mRNA相对表达量显著高于空白对照组 (P<0.05)；2组细胞中Bcl-2、Mcl-1 mRNA相对表达量比较差异无统计学意义 (P>0.05)。结论　腺苷可通过诱导A549细胞周期阻滞和凋亡来抑制细胞增殖能力，腺苷诱导A549细胞凋亡与其上调促凋亡因子Bak基因表达从而改变线粒体膜通透性有关。腺苷还可抑制A549细胞迁移能力。
Abstract:
Objective　To investigate the effect of adenosine on proliferation of lung cancer cells and the potential molecular mechanism.Methods　The A549 cells in logarithmic growth phase were taken and divided into the blank control group and 0.5,1.0,2.5,5.0 mmol·L-1adenosine treatment group;the above five groups were added 0.0,0.5,1.0,2.5,5.0 mmol·L-1adenosine medium for culture.The effect of adenosine on A549 cells proliferation was detected by cell counting kit-8,the cell cycle and cell apoptosis were examined by flow cytometry,the cells migration capacity was detected by scratch assay,the expression of apoptosis related molecular was detected by quantitative real-time polymerase chain reaction.Results　The growth inhibition rate of A549 cells in the 0.5,1.0,2.5,5.0 mmol·L-1adenosine treatment group was signi-ficantly higher than that in the blank control group at 24,48 h after treatment(P<0.05);the growth inhibition rate of A549 cells in different concentrations of adenosine treatment groups increased with the increase of the concentration of adenosine at 24,48 h after treatment,and the difference was statistically significant (P<0.05).The relative migration rates of A549 cells in 2.5 mmol·L-1 and 5.0 mmol·L-1 adenosine treatment groups were significantly lower than those in the blank control group (P<0.05);the relative migration rate of A549 cells in 5.0 mmol · L-1 adenosine treatment group was significantly lower than that in the 2.5 mmol·L-1 adenosine treatment group (P<0.05).Compared with the blank control group,the proportion of A549 cells in G0/G1 phase decreased and in S，G2/M phase increased in the 2.5 mmol·L-1 adenosine treatment group (P<0.05)；the proportion of A549 cells in G0/G1 phase decreased in G2/M phase increased in the 5.0 mmol·L-1 adenosinetreatment group (P<0.05);there was no significant difference in the proportion of A549 cells in S phase between the blank control group and the 5.0 mmol·L-1 adenosine treatment group(P>0.05).Compared with the 2.5 mmol·L-1 adenosine treatment group,the proportion of A549 cells in S phase decreased,while that in G2/M phase increased in the 5.0 mmol·L-1 adenosine treatment group (P<0.05);there was no significant difference in the proportion of A549cells in G0/G1 phase between the 2.5 mmol·L-1 adenosine treatment group and 5.0 mmol · L-1 adenosine treatment group (P>0.05).The apoptosis rates of A549 cells in the 2.5 mmol·L-1 and 5.0 mmol·L-1 adenosine treatment group were significantly higher than those in the blank control group (P<0.05);the apoptosis rate of A549 cells in the 5.0 mmol·L-1 adenosine treatment group was significantly higher than that in the 2.5 mmol·L-1 adenosine treatment group (P<0.05).The relative expression of Bak,Caspase-3 mRNA in A549 cells in the 5.0 mmol·L-1 adenosine treatment group was significantly higher than that in the blank control group(P<0.05);there was no significant difference in the relative expression of Bcl-2 and Mcl-2 mRNA in A549 cells between the two groups (P>0.05).Conclusion　Adenosine can inhibit the proliferation of A549 cells by inducing cell apoptosis and cell cycles arrest.Adenosine induce the apoptosis of A549 cells was associated with the up-regulation of Bak gene expression.Adenosine also can inhibite the migration of A549 cells.

参考文献/References:

［1］　SIEGEL R L,MILLER K D,JEMAL A.Cancer statistics,2019［J］.CA Cancer J Clin，2019,69(1):7-34.
［2］　SIEGEL R L,MILLER K D,JEMAL A.Cancer statistics,2020［J］.CA Cancer J Clin，2020,70(1):7-30.
［3］　ANTONIOLI L,FORNAI M,BLANDIZZI C,et al.Adenosine signaling and the immune system:when a lot could be too much［J］.Immunol Lett,2019,205(1):9-15.
［4］　BESSA-GONCALVES M,BRAGANCA B,MARTINS-DIAS E,et al.Is the adenosine A2B ′biased′ receptor a valuable target for the treatment of pulmonary arterial hypertension［J］.Drug Discov Today,2018,23(6):1285-1292.
［5］　CHEN M,WEI L,LAW C T,et al.RNA N6-methyladenosine methyltransferase-like 3 promotes liver cancer progression through YTHDF2-dependent posttranscriptional silencing of SOCS2［J］.Hepatology,2018,67(6):2254-2270.
［6］　MORANDI F,HORENSTEIN A L,RIZZO R,et al.The role of extracellular adenosine generation in the development of autoimmune diseases［J］.Mediators Inflamm,2018,2018:7019398.
［7］　YANG D,YAGUCHI T,YAMAMOTO H,et al.Intracellularly transported adenosine induces apoptosis in HuH-7 human hepatoma cells by downregulating c-FLIP expression causing caspase-3/-8 activation［J］.Biochem Pharmacol,2007,73(10):1665-1675.
［8］　MUJOOMDAR M,HOSKIN D,BLAY J.Adenosine stimulation of the proliferation of colorectal carcinoma cell lines.Roles of cell density and adenosine metabolism［J］.Biochem Pharmacol,2003,66(9):1737-1747.
［9］ 　高文强，刘坤，周艳，等.腺苷对人胃癌MGC-803细胞增殖与凋亡的影响［J］.实用临床医药杂志，2016，20(13)：44-47，55.
［10］　SAITO M,YAGUCHI T,YASUDA Y,et al.Adenosine suppresses CW2 human colonic cancer growth by inducing apoptosis via A(1) adenosine receptors［J］.Cancer Lett,2010,290(2):211-215.
［11］　SRDIC D,PLESTINA S,SVERKO-PETERNAC A,et al.Cancer cachexia,sarcopenia and biochemical markers in patients with advanced non-small cell lung cancer-chemotherapy toxicity and prognostic value.Supportive care in cancer［J］.Support Care Cancer,2016,24(11):4495-4502.
［12］　CHEN J F,ELTZSCHIG H K,FREDHOLM B B.Adenosine receptors as drug targets：what are the challenges［J］.Nat Rev Drug Discov,2013,12(4):265-286.
［13］　ELTZSCHIG H K.Extracellular adenosine signaling in molecular medicine［J］.J Mol Med (Berl),2013,91(2):141-146.
［14］　SHIRALI S,AGHAEI M,SHABANI M,et al.Adenosine induces cell cycle arrest and apoptosis via cyclin D1/Cdk4 and Bcl-2/Bax pathways in human ovarian cancer cell line OVCAR-3［J］.Tumour Biol,2013,34(2):1085-1095.
［15］　JAFARI S M,JOSHAGHANI H R,PANJEHPOUR M,et al.Apoptosis and cell cycle regulatory effects of adenosine by modulation of GLI-1 and ERK1/2 pathways in CD44+ and CD24- breast cancer stem cells［J］.Cell Prolif,2017,50(4):e12345.
［16］　王会平，郜赵伟，刘冲，等.腺苷抑制乳腺癌细胞增殖和迁移的机制［J］.现代肿瘤医学，2016，24(9)：1349-1352.
［17］　MA Y,ZHANG J,ZHANG Q,et al.Adenosine induces apoptosis in human liver cancer cells through ROS production and mitochondrial dysfunction［J］.Biochem Biophys Res Commun,2014,448(1):8-14.
［18］　TSUCHIYA A,NISHIZAKI T.Anticancer effect of adenosine on gastric cancer via diverse signaling pathways［J］.World J Gastroenterol,2015,21(39):10931-10935.
［19］　YU S,HOU D,CHEN P,et al.Adenosine induces apoptosis through TNFR1/RIPK1/P38 axis in colon cancer cells［J］.Biochem Biophys Res Commun,2015,460(3):759-765.
［20］　NUTT L K,PATAER A,PAHLER J,et al.Bax and Bak promote apoptosis by modulating endoplasmic reticular and mitochondrial Ca2+ stores［J］.J Biol Chem,2002,277(11):9219-9225.
［21］　BAUER J H,HELFAND S L.New tricks of an old molecule:lifespan regulation by p53［J］.Aging Cell,2006,5(5):437-440.
［22］　SAMUELS-LEV Y,O′CONNOR D J,BERGAMASCHI D,et al.ASPP proteins specifically stimulate the apoptotic function of p53［J］.Mol Cell,2001,8(4):781-794.
［23］　方慧华，李洪娇，何小蕾，等.小檗碱及其衍生物抗肿瘤作用研究进展［J］.中国药学杂志，2017，52(6)：429-433.
［24］　邹翔，曲中原，季宇彬.胡桃醌经由活性氧介导JNK、p38通路诱导SGC-7901细胞凋亡［J］.中国药学杂志，2016，51(7)：544-549.
［25］　GAO Z W,WANG H P,DONG K,et al.Adenosine inhibits migration,invasion and induces apoptosis of human cervical cancer cells［J］.Neoplasma,2016,63(2):201-207.

相似文献/References:

[1]路平,梁秋冬,郑全庆.宫颈鳞癌中erbB3、erbB4基因蛋白表达与细胞凋亡和增殖的相关性研究[J].新乡医学院学报,2003,20(05):317.
[2]张卫国。,董果雄,韩小宁,等.全反式维甲酸对培养大鼠主动脉血管平滑肌细胞增殖及细胞周期素E表达的影响[J].新乡医学院学报,2003,20(05):322.
[3]路　平,梁秋冬,郑全庆.宫颈鳞癌中erbB3 、erbB4 基因蛋白表达与细胞凋亡和增殖的相关性研究[J].新乡医学院学报,2003,20(05):317.
[4]张卫国,董果雄,韩小宁,等.全反式维甲酸对培养大鼠主动脉血管平滑肌细胞增殖及细胞周期素E表达的影响[J].新乡医学院学报,2003,20(05):322.
[5]王　峰,张　煜,李娜娜.碱性成纤维细胞生长因子对小鼠骨髓内皮细胞增殖的促进作用[J].新乡医学院学报,2006,23(05):475.
[6]王蕾,赵玉林,苗旺,等.老年性耳聋患者内皮祖细胞功能变化 TitleFilter('chTitle');[J].新乡医学院学报,2010,27(04):348.
[7]徐建可,栗延伟,韩新生,等.星形胶质细胞条件培养液对缺氧损伤神经元的影响[J].新乡医学院学报,2012,29(10):750.
[8]刘莹莹,刘晶晶,刘志华.外加直流电场对脑卒中大鼠侧脑室室管膜下区神经干细胞增殖与迁移的影响[J].新乡医学院学报,2015,32(12):1086.
[9]王　静,康媛洁,王　贞,等.DEAH盒解旋酶15对转化生长因子-β1诱导的呼吸道平滑肌细胞增殖和迁移的影响[J].新乡医学院学报,2021,38(5):413.[doi:10.7683/xxyxyxb.2021.05.003]
　WANG Jing,KANG Yuanjie,WANG Zhen,et al.Effect of DEAH-box helicase 15 on the proliferation and migration of airway smooth muscle cells induced by transforming growth factor-β1[J].Journal of Xinxiang Medical University,2021,38(8):413.[doi:10.7683/xxyxyxb.2021.05.003]
[10]段　聿,陈　吉,崔　宏,等.长链非编码RNA尿路上皮癌胚抗原1/微RNA-143-3p/成纤维细胞生长因子1轴对胃癌细胞BGC-823增殖、迁移及侵袭的影响[J].新乡医学院学报,2021,38(7):607.[doi:10.7683/xxyxyxb.2021.07.002]
　DUAN Yu,CHEN Ji,CUI Hong,et al.Effect of long non-coding RNA urothelial carcinoma antigen 1/microRNA-143-3p/fibroblast growth factor 1 axis on proliferation,migration and invasion of gastric cancer BGC-823 cell[J].Journal of Xinxiang Medical University,2021,38(8):607.[doi:10.7683/xxyxyxb.2021.07.002]

更新日期/Last Update: 2020-08-05