基础医学与临床 ›› 2015, Vol. 35 ›› Issue (10): 1314-1319.

• 研究论文 • 上一篇    下一篇

STAT3异常激活对人脐带间充质干细胞在乳腺癌微环境中恶性转化的影响

汪玲,朱静,田杰,谭彬,燕莎   

  1. 重庆医科大学附属儿童医院
  • 收稿日期:2015-04-27 修回日期:2015-06-30 出版日期:2015-10-05 发布日期:2015-09-30
  • 通讯作者: 朱静 E-mail:zhujing310@yahoo.com.cn
  • 基金资助:
    国家自然科学基金面上项目

Effects of aberrant activation of STAT3 in malignant transformation of human umbilical correlation cord mesenchymal stem cells when exposed to human breast cancer microenvironment

  • Received:2015-04-27 Revised:2015-06-30 Online:2015-10-05 Published:2015-09-30
  • Contact: Jing ZHU E-mail:zhujing310@yahoo.com.cn

摘要: 目的 探讨人脐带间充质干细胞(hUCMSCs)在MCF-7B乳腺癌微环境中是否存在恶性转变并探究其生物学变化与STAT3异常激活及高表达的关系。 方法 将hUCMSCs分为3组:空白对照组(hUCMSCs单独培养)、实验组(hUCMSCs与MCF-7B共培养)、阳性对照组(MCF-7B单独培养)。倒置显微镜观察细胞形态变化;流式细胞术检测细胞周期;RT-qPCR检测STAT3及其下游原癌基因c-Myc及抗凋亡Bcl-xLmRNA的表达;细胞免疫荧光检测p-STAT3、c-Myc和Bcl-xL蛋白表达及定位;Western blot检测p-STAT3、STAT3、c-Myc和Bcl-xL蛋白表达。结果 实验组hUCMSCs与对照组比较核质比增大,细胞大小不一 ,排列紊乱;实验组细胞G1期比例显著低于对照组(P<0.05),S期和G2期比例均显著高于对照组(P<0.05);实验组STAT3、c-Myc和Bcl-xLmRNA的表达均显著高于对照组(P<0.05);实验组p-STAT3、STAT3、c-Myc和Bcl-xL的蛋白表达水平显著高于对照组(P<0.05),且主要定位于细胞核内。结论hUCMSCs在MCF-7B乳腺癌微环境中存在恶性转化趋势,且STAT3的异常激活及高表达是hUMSCs恶性转化的重要因素之一。

关键词: 人脐带间充质干细胞, 恶性转化, 信号传导与转录活化因子3

Abstract: Abstract:Objiective To investigate whether hUCMSCs undergo malignant transformation when exposed to MCF-7B breast cancer microenvironment and whether the abnormal activation and over expression of STAT3 play an important role in this transformation. Methods The experiment was divided into three groups:blank group(hUCMSCs were separately cultured),experimental group(hUCMSCs were indirectly co-cultured with MCF-7B breast cancer cells),positive control group(MCF-7B breast cancer cells were separately cultured ).Morphology of cells was detected by invertedmicroscope. Cell cycle was detected by Flow cytometry. The mRNA expressions of STAT3、c-Myc and Bcl-xL were tested by real-time PCR.The protein expressions and locations of p-STAT3、c-Myc and Bcl-xL were detected by immunofluorescence.The protein expressions of p-STAT3、STAT3、c-Myc and Bcl-xL were also measured by Western blot.Results The experimental group cells showed typical morphology features of tumor cell. The cells proportion of experiment group in G1 phase was significantly lower than that of the blank group(P<0.05),but which in S and G2 phase were significantly higher than those of the blank group(P<0.05).The mRNA expression levels of STAT3、c-Myc and Bcl-xL in experimental group were significantly higher than those in blank group(P<0.05). p-STAT3、STAT3、c-Myc and Bcl-xL protein were significantly higher than that of the blank group(P<0.05),and they were mainly situated in the nuclei.The protein expressions of STATS also showed significant changes in experimental group. Conclusions hUCMSCs trends to malignant transformations when exposed to MCF-7B breast cancer microenvironment.And the abnormal activation and over expression of STAT3 is one of important factors leading to the malignant transformation of hUCMSCs.

Key words: human Umbilical Cord Mesenchymal Stem Cells, malignant transformation, Signal transducerand activator of transcription 3

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