[1]郭　满,翟晓建,张　浩,等.p38蛋白激酶信号通路对骨桥蛋白介导的三阴性乳腺癌细胞自噬、凋亡活性的影响[J].新乡医学院学报,2018,35(12):1058-1063.[doi:10.7683/xxyxyxb.2018.12.004]
　GUO Man,ZHAI Xiao-jian,ZHANG Hao,et al.Effect of p38 mitogen-activated protein kinase signaling pathway on autophagy and apoptosis activity of triple negative breast cancer cell mediated by osteopontin[J].Journal of Xinxiang Medical University,2018,35(12):1058-1063.[doi:10.7683/xxyxyxb.2018.12.004]
点击复制

p38蛋白激酶信号通路对骨桥蛋白介导的三阴性乳腺癌细胞自噬、凋亡活性的影响
分享到：

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

卷:
35
期数:
2018年12
页码:
1058-1063
栏目:
基础研究
出版日期:
2018-12-05

文章信息/Info

Title:
Effect of p38 mitogen-activated protein kinase signaling pathway on autophagy and apoptosis activity of triple negative breast cancer cell mediated by osteopontin
作者:
郭　满1; 翟晓建1; 张　浩1; 李伟汉1; 刘　明2
（1.南阳市中心医院乳腺外科，河南　南阳　473003；2.新乡医学院研究生院，河南　新乡　453003）
Author(s):
GUO Man1; ZHAI Xiao-jian1; ZHANG Hao1; LI Wei-han1; LIU Ming2
（1.Department of Galactophore Surgery，the Central Hospital of Nanyang City，Nanyang 473003，Henan Province，China；2.Graduate School of Xinxiang Medical University，Xinxiang 453003，Henan Province，China）
关键词:
三阴性乳腺癌; 骨桥蛋白; 自噬; p38蛋白激酶; Beclin 1
Keywords:
triple negative breast cancer; osteopontin; autophagy; p38 mitogen-activated protein kinase; Beclin 1
分类号:
R737.9
DOI:
10.7683/xxyxyxb.2018.12.004
文献标志码:
A
摘要:
目的　探讨骨桥蛋白（OPN）和自噬相关蛋白Beclin 1在三阴性乳腺癌组织中的表达及OPN下调对p38蛋白激酶（p38MAPK）、Beclin 1表达及乳腺癌细胞自噬、凋亡活性的影响。方法　选取2015年1月至2016年12月南阳市中心医院收治的60例女性三阴性乳腺癌患者的癌组织标本为研究对象，另选取同期手术治疗的40例女性乳腺纤维腺瘤或乳腺增生结节患者的正常乳腺组织作为对照；采用免疫组织化学方法检测三阴性乳腺癌组织和正常乳腺组织中OPN和Beclin 1的表达。将MDA-MB-231细胞株分为特异性OPN小干扰RNA（siRNA）实验组（OPN-siＲNA组）、非同源阴性CON-siRNA对照组（CON-siRNA组）和mock空白对照组（mock组）；以慢病毒介导基因干扰技术制备OPN低表达MDA-MB-231细胞模型，应用透射电镜及流式细胞术检测OPN下调后细胞自噬体形成和凋亡率，实时荧光定量聚合酶链反应（PCR）和Western blot法检测OPN下调后p38MAPK及Beclin 1的表达。结果　OPN蛋白在乳腺癌组织和正常乳腺组织中的阳性表达率分别为68.3％（41/60）和12.5％（5/40），OPN蛋白在乳腺癌组织中的阳性表达率高于正常乳腺组织（χ2=56.881，P=0.000）；Beclin 1蛋白在乳腺癌组织和正常乳腺组织中的阳性表达率分别为18.3％（11/60）和42.5％（17/40），Beclin 1蛋白在乳腺癌组织中的阳性表达率低于正常乳腺组织（χ2=54.692，P=0.000）；三阴性乳腺癌组织中OPN蛋白表达与Beclin 1蛋白表达呈负相关（r=-0.713，P=0.003）。病毒转染48 h后， OPN-siRNA组细胞中OPN mRNA和蛋白相对表达量显著低于CON-siRNA组和mock组（F=57.170、50.410，P=0.001）。OPN 表达下调后，OPN-siRNA组细胞中自噬体个数、早期凋亡率显著高于CON-siRNA组和mock组（F=17.590，P=0.004；F=23.180，P=0.003）；OPN-siRNA组细胞中Beclin 1 mRNA和蛋白相对表达量明显高于CON-siRNA组和mock组（F=29.873、32.174，P=0.001），p38MAPK mRNA和蛋白相对表达量低于 mock组和CON-siRNA组（F=32.735、37.110，P=0.000）；mock组与CON-siRNA组细胞中自噬体个数、早期凋亡率、Beclin 1 mRNA和蛋白相对表达量及p38MAPK mRNA和蛋白相对表达量比较差异均无统计学意义（P>0.05）。结论　抑制OPN表达可促进三阴性乳腺癌细胞自噬和凋亡活性，其作用机制可能与p38MAPK表达下调、Beclin 1表达上调有关。
Abstract:
Objective　To study the expression of osteopontin （OPN） and autophagy re-lated protein Beclin 1 in triple negative breast cancer tissues，and to explore the effect of OPN knockdown on the expression of p38 mitogen-activated protein kinase（p38MAPK）、Beclin 1 and autophagy，apoptosis of breast cancer cells.Methods　A total of 60 female patients with triple negative breast cancer admitted to the the Central Hospital of Nanyang City from January 2015 to December 2016 were selected as study objects，and forty normal breast tissue specimens of female patients with breast fibroadenoma or breast hyperplasia nodules treated at the same time were selected as the control.Immunohistochemistry were used to detect the expression of OPN and Beclin 1 in the triple negative breast cancer tissues and normal breast tissues.MDA-MB-231 cell lines were divided into specific OPN small interference RNA experimental group （OPN-siRNA group），the same negative CON-siRNA control group （CON-siRNA group） and mock blank control group （mock group）.Lentivirus mediated RNA interfer-ence technology were employed to build OPN low expression MDA-MB-231cells，then transmission electron microscope and flow cytometry were used to detect auto-phagy and apoptosis of MDA-MB-231 cells after OPN knockdown，western blot and real-time fluorescence quantitative polymerase chain reaction were used to detect the expression of p38MAPK and Beclin 1.Results　The positive expression rate of OPN protein in breast cancer tissue and normal breast tissue was 68.3% （41/60） and 12.5% （5/40），the positive ex-pression rate of OPN protein in breast cancer tissue was higher than that in the normal breast tissue （χ2=56.881，P=0.000）.The positive expression rate of Beclin 1 protein in breast cancer tissues and normal breast tissues was 18.3% （11/60） and 42.5% （17/40） respectively，and the positive expression rate of Beclin 1 protein in breast cancer tissues was lower than that in normal breast tissue （χ2=54.692，P=0.000）.OPN protein expression in tri-negative breast cancer tissues was negatively correlated with Beclin 1 protein expression （r=-0.713，P=0.003）.After 48 h viral transfection，the relative expression of OPN mRNA and protein in the OPN-siRNA group was significantly lower than that in the CON-siRNA group and the mock group （F=57.170，50.410，P=0.001）.After the down-regulation of OPN expression，the number of autophagosomes and early apoptosis rate in cells of the OPN-siRNA group were significantly higher than those of the CON-siRNA group and the mock group （F=17.590，P=0.004；F=23.180，P=0.003）.In the OPN-siRNA group，the relative expression of Beclin 1 mRNA and protein was significantly higher than that of the CON-siRNA group and the mock group （F=29.873，32.174，P=0.001），and the relative expression of p38MAPK mRNA and protein was lower than that of the mock group and the CON-siRNA group （F=32.735，37.110，P=0.000）.There was no statistically significance in the number of autophagosomes，early apoptosis rate，relative expression of Beclin 1 mRNA and protein，and relative expression of p38MAPK mRNA and protein in cells between the mock group and the CON-siRNA group （P>0.05）.Conclusion　Inhibition of OPN expression could promote autophagy and apoptosis of triple negative breast cancer cells，the underlying mechanism of this phenomenon may be related to down-regulation of p38MAPK and up-regulation of Beclin 1.

参考文献/References:

［1］　DESANTIS C，MA J，BRYAN L，et al.Breast cancer statistics，2013［J］.CA Cancer J Clin，2014，64（1）：52-62.
［2］　DIETZE E C，SISTRUNK C，MIRANDA-CARBONI G，et al.Triple-negative breast cancer in African-American women：disparities versus biology［J］.Nat Rev Cancer，2015，15（4）：248-254.
［3］　ZDUNIAK K，ZIOLKOWSKI P，AHLIN C，et al.Nuclear osteopontin-c is a prognostic breast cancer marker［J］.Br J Cancer，2015，112（4）：729-738.
［4］　GU Y，LI P，PENG F，et al.Autophagy-related prognostic signature for breast cancer［J］.Mol Carcinog，2016，55（3）：292-299.
［5］　HABIB R，AKHTAR J，TAQI M，et al.Lentiviral vector-mediated survivin shRNA delivery in gastric cancer cell lines significantly inhibits cell proliferation and tumor growth［J］.Oncol Rep，2015，34（2）：859-867.
［6］　董树旭，赵轼轩，王颖，等.利用透射电镜技术分析血细胞自噬在血液病中的分布特点［J］.中华血液学杂志，2015，36（2）：144-147.
［7］　秦海运，潘淑云，李政，等.复方苦参注射液联合拉帕替尼治疗Her-2阳性晚期乳腺癌疗效及对肿瘤标志物的影响［J］.世界中医药，2018，13（6）：1472-1476.
［8］　YANG L，WEI L，ZHAO W，et al.Down-regulation of osteopontin expression by RNA interference affects cell proliferation and che-motherapy sensitivity of breast cancer MDA-MB-231 cells［J］.Mol Med Rep，2012，5（2）：373-376.
［9］　ORTIZ-MARTINEZ F，SANMARTIN E，POMARES-NAVARRO E，et al.Osteopontin regulates VEGFA and ICAM-1 mRNA expression in breast carcinoma［J］.Am J Clin Pathol，2015，143（6）：812-822.
［10］　焦庆丽，吴爱国，邵国利，等.乳腺癌组织Beclin 1和EMT相关分子标志物表达及其临床意义探讨［J］.中华肿瘤防治杂志，2015，22（18）：1456-1460.
［11］　许杜娟，杨翠，王猛武，等.保护性自噬对顺铂诱导人乳腺癌MCF-7细胞凋亡的抑制作用探讨［J］.安徽医药，2015,19（1）：152-155.
［12］　YU X，LI R，SHI W，et al.Silencing of microRNA-21 confers the sensitivity to tamoxifen and fulvestrant by enhancing autophagic cell death through inhibition of the PI3K-AKT-mTOR pathway in breast cancer cells［J］.Biomed Pharmacother，2016，361（77）：37-44.
［13］　JUNG Y Y，LEE Y K，KOO J S.The potential of Beclin 1 as a therapeutic target for the treatment of breast cancer［J］.Expert Opin Ther Targets，2016，20（2）：167-178.
［14］　LI J P，YANG Y X，LIU Q L，et al.The investigational aurora kinase A inhibitor alisertib （MLN8237） induces cell cycle G2/M arrest，apoptosis，and autophagy via p38 MAPK and Akt/mTOR signaling pathways in human breast cancer cells［J］.Drug Des Devel Ther，2015，9（9）：1627-1652.

相似文献/References:

[1]路　娜,古吉敏,闵志波.肿瘤间质比在三阴性乳腺癌患者预后评估中的价值[J].新乡医学院学报,2017,34(12):1089.[doi:10.7683/xxyxyxb.2017.12.011]
　LU Na,GU Ji-min,MIN Zhi-bo.Value of tumor-stroma ratio in evaluating the prognosis of patients with triple-negative breast cancer[J].Journal of Xinxiang Medical University,2017,34(12):1089.[doi:10.7683/xxyxyxb.2017.12.011]
[2]高金亮,赵　楠,张　浩,等.乳腺癌患者血清及癌组织中骨桥蛋白表达及意义[J].新乡医学院学报,2019,36(6):585.[doi:10.7683/xxyxyxb.2019.06.022]
　GAO Jin-liang,ZHAO Nan,ZHANG Hao,et al.Significance of the expression of osteopontin in serum and cancer tissues of patients with breast cancer[J].Journal of Xinxiang Medical University,2019,36(12):585.[doi:10.7683/xxyxyxb.2019.06.022]
[3]刘德总,杨颖涛.乳腺癌辅助治疗现状与展望[J].新乡医学院学报,2021,38(8):781.[doi:10.7683/xxyxyxb.2021.08.017]
[4]郭彬彬,郭　满,刘　明,等.扭曲蛋白、波形蛋白、血管生成拟态在三阴性乳腺癌组织中的表达及临床意义[J].新乡医学院学报,2021,38(5):436.[doi:10.7683/xxyxyxb.2021.05.008]
　GUO Binbin,GUO Man,LIU Ming,et al.Expression of Twist,vimentin and vasculogenic mimicry in triple-negative breast cancer tissues and its clinical significance[J].Journal of Xinxiang Medical University,2021,38(12):436.[doi:10.7683/xxyxyxb.2021.05.008]
[5]刘　韡,杨　欢,彭慧霞,等.复方苦参注射液联合化学治疗和调强适形放射治疗中晚期宫颈癌疗效观察[J].新乡医学院学报,2020,37(12):1156.[doi:10.7683/xxyxyxb.2020.12.012]
　LIU Wei,YANG Huan,PENG Huixia,et al.Effect of Fufang Kushen Zhusheye combined with chemotherapy and intensity modulated radiation therapy in treatment of advanced cervical cancer[J].Journal of Xinxiang Medical University,2020,37(12):1156.[doi:10.7683/xxyxyxb.2020.12.012]
[6]王小雯.血清骨桥蛋白、神经胶质纤维酸性蛋白及生长分化因子15水平在急性脑梗死患者神经损伤及预后评估中的应用价值[J].新乡医学院学报,2019,36(5):479.[doi:10.7683/xxyxyxb.2019.05.019]
　WANG Xiao-wen.Application value of serum osteopontin，neurofibrillar acidic protein and growth differentiation factor 15 in the assessment of nerve injury and prognosis in patients with acute cerebral infarction[J].Journal of Xinxiang Medical University,2019,36(12):479.[doi:10.7683/xxyxyxb.2019.05.019]
[7]付朝红,刘丹娜,段方方,等.晚期三阴性乳腺癌核苷酸切除修复交叉互补基因1表达与化学治疗效果的关系[J].新乡医学院学报,2017,34(10):904.[doi:10.7683/xxyxyxb.2017.10.010]
　FU Chao-hong,LIU Dan-na,DUAN Fang-fang,et al.Relationship between excision repair cross complementing gene 1 expression and chemotherapeutic efficacy in patients with advanced triple-negative breast cancer[J].Journal of Xinxiang Medical University,2017,34(12):904.[doi:10.7683/xxyxyxb.2017.10.010]
[8]朱方园,邵营波,陈　琦,等.长链非编码RNA前列腺癌相关转录本7对三阴性乳腺癌MDA-MB-436细胞增殖、迁移及侵袭的影响[J].新乡医学院学报,2022,39(8):701.[doi:10.7683/xxyxyxb.2022.08.001]
　ZHU Fangyuan,SHAO Yingbo,CHEN Qi,et al.Effect of long non-coding RNA prostate cancer associated transcript 7 on proliferation,migration and invasion of triple-negative breast cancer MDA-MB-436 cells[J].Journal of Xinxiang Medical University,2022,39(12):701.[doi:10.7683/xxyxyxb.2022.08.001]
[9]马新豫.血清骨桥蛋白和D-二聚体水平对心肌梗死介入治疗患者预后的影响[J].新乡医学院学报,2022,39(11):1047.[doi:10.7683/xxyxyxb.2022.11.009]
　MA Xinyu.Effect of serum osteopontin and D-dimer levels on prognosis of patients with myocardial infarction undergoing interventional therapy[J].Journal of Xinxiang Medical University,2022,39(12):1047.[doi:10.7683/xxyxyxb.2022.11.009]
[10]吴昊,段海明.人表皮生长因子受体2阳性和三阴性乳腺癌患者新辅助化疗后腋窝淋巴结病理学分析[J].新乡医学院学报,2024,(4):358.[doi:10.7683/xxyxyxb.2024.04.011]
　WU Hao,DUAN Haiming.Pathology of axillary lymph nodes after neoadjuvant chemotherapy for human epidermal growth factor receptor 2-positive and triple-negative breast cancer patients[J].Journal of Xinxiang Medical University,2024,(12):358.[doi:10.7683/xxyxyxb.2024.04.011]
[11]刘　明,郭　满,张　浩,等.血管生成拟态及骨桥蛋白和基质金属蛋白酶2在三阴性乳腺癌组织中的表达及意义[J].新乡医学院学报,2019,36(4):345.[doi:10.7683/xxyxyxb.2019.04.011]
　LIU Ming,GUO Man,ZHANG Hao,et al.Expression and significance of vasculogenic mimicry，osteopontin and matrix metalloproteinase-2 in triple-negative breast cancer[J].Journal of Xinxiang Medical University,2019,36(12):345.[doi:10.7683/xxyxyxb.2019.04.011]

更新日期/Last Update: 2018-12-05