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缺氧条件下人脐带源性间充质干细胞增强缺氧诱导因子-1α表达促进肺腺癌细胞增殖

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缺氧条件下人脐带源性间充质干细胞增强缺氧诱导因子-1α表达促进肺腺癌细胞增殖
Upregulation of hypoxia-inducible factor-1α by human umbilical cord-derived mesenchymal stem cells under hypoxia facilitates the proliferation of lung adenocarcinoma cells
上传时间:2019/7/5 14:54:14    作者:李昶,赵成岭,陈诗军,陈余清|下载

李昶,赵成岭,陈诗军,陈余清*
(蚌埠医学院第一附属医院呼吸与危重症学科,蚌埠233004)

Li Chang, Zhao Chengling, Chen Shijun, Chen Yuqin*
(Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China)
摘要:目的 探讨缺氧条件下缺氧诱导因子-1α (hypoxia-inducible factor-1α, HIF-1α)在人脐带源性间充质干细胞(human umbilical cord-derived mesenchymal stem cells, hucMSCs)促进肺腺癌A549细胞增殖过程中的可能机制。方法 分离培养和鉴定hucMSCs,分别与肺腺癌A549细胞、干扰了HIF-1α基因的A549细胞在常氧或缺氧条件下进行共培养,荧光定量PCR和Western blot法分别检测HIF-1α和生存素(survivin)表达水平,MTT法检测细胞增殖力,划痕实验分析细胞迁移力。结果 MTT和划痕实验检测显示,常氧条件下,hucMSCs共培养的A549 细胞增殖能力、迁移能力较无hucMSCs共培养的细胞明显减弱;缺氧条件下,A549 细胞增殖力和迁移力较常氧培养的A549 细胞明显增强,hucMSCs共培养使二者进一步增强,沉默HIF-1α则使二者减弱,hucMSCs共培养不改变沉默HIF-1α对细胞增殖力和迁移力的影响。荧光定量PCR和Western blot检测显示,缺氧培养时,A549 细胞内HIF-1α和survivin表达明显上调,hucMSCs共培养明显增强缺氧对HIF-1α和survivin表达的上调作用;沉默HIF-1α抑制缺氧对A549细胞HIF-1α和survivin表达的上调,hucMSCs共培养不改变沉默HIF-1α对HIF-1α和survivin表达上调的抑制。结论  缺氧条件下,hucMSCs共培养导致肺腺癌A549细胞HIF-1α和survivin表达上调,从而使A549细胞的增殖和迁移能力增强。
关键词:肺腺癌;缺氧;人脐带源性间充质干细胞;缺氧诱导因子-1α;生存素
Abstract: Objective To investigate the possible mechanism underlying the promotion effect of hypoxia-inducible factor-1α (HIF-1α) on the proliferation of lung adenocarcinoma cells under the regulation of human umbilical cord-derived mesenchymal stem cells (hucMSCs) under hypoxia. Methods After isolated culture and identification, hucMSCs were co-cultured with lung adenocarcinoma A549 cells or A549 cells with interfering HIF-1α gene under normoxia or hypoxia, respectively. Subsequently, quantitative real-time PCR and western blot were used to detect the expression level of HIF-1α and survivin in A549 cells cultured alone or co-cultured with hucMSCs, before or after interfering HIF-1α gene, under normoxia or hypoxia, respectively. MTT assay was used to detect the proliferation of A549 cells. Scratch assay was uesd to analyse the ability of migration of A549 cells. Results MTT assay and Scratch assay showed that poliferation and migration of A549 cells co-cultured with hucMSCs decreased significantly compared to A549 without co-culture under normoxia. Under hypoxia, the proliferation and migration of A549 cells were significantly enhanced compared with those in normoxia, which were further enhanced when co-cultured with hucMSCs. However, silencing HIF-1α weakened both the proliferation and migration, and the co-culture with hucMSCs did not change the effects of silencing HIF-1α on cell proliferation and migration. Quantitative real-time PCR and western blot showed that the expression of HIF-1α and survivin was up-regulated in A549 cells during hypoxia culture, which was significantly enhanced due to the co-culture of hucMSCs but was inhibited by silencing of HIF-1α. The co-culture with hucMSCs did not change the inhibition of silencing HIF-1α. Conclusion Under hypoxic conditions, co-culture with hucMSCs up-regulated the expression of HIF-1α and survivin in lung adenocarcinoma A549 cells, which enhanced the proliferation and migration of A549 cells.
Keywords: Lung adenocarcinoma cells; hypoxia; human umbilical cord-derived mesenchymal stem cells; hypoxia inducible factor-1α; HIF-1α; survivin

〔中图分类号〕R734.2             〔文献标识码〕A             DOI:10.16705/ j. cnki. 1004-1850. 2019. 02. 002

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