环孢素A调控自噬诱导肿瘤相关巨噬细胞极化干预子宫内膜癌细胞的恶性生物学行为

被引:2
作者
徐丽
程欣
王秀虹
钟晖
马丹
机构
[1] 咸宁市中心医院(湖北科技学院附属第一医院)妇科
关键词
环孢素A; 子宫内膜癌; 肿瘤相关巨噬细胞; 自噬;
D O I
暂无
中图分类号
R737.33 [子宫肿瘤];
学科分类号
摘要
目的:探究环孢素A(CsA)对肿瘤相关巨噬细胞极化的调控作用及其与子宫内膜癌(EC)细胞恶性生物学行为之间的联系,并初步探讨相关机制。方法:采用佛波酯(PMA)和白细胞介素-4(IL-4)诱导THP-1细胞向具有肿瘤相关巨噬细胞极化分型特点的M2样巨噬细胞分化,分别以50、100、150、200 ng/mL CsA处理诱导后的细胞24 h, CCK-8法检测细胞活性,流式细胞术检测CD86和CD206表达,实时荧光定量聚合酶链式反应(qRT-PCR)检测诱导型一氧化氮合酶(iNOS)、肿瘤坏死因子-α(TNF-α)、转化生长因子-β(TGF-β)、重组人精氨酸酶1(Arg-1)的mRNA相对表达量。将人子宫内膜癌Ishikawa细胞随机分为4组并进行相应处理:对照组,Ishikawa细胞加入细胞培养液培养24 h; CsA组,200 ng/mL CsA干预Ishikawa细胞24 h;肿瘤相关巨噬细胞(TAM)组,肿瘤相关巨噬细胞培养液上清干预Ishikawa细胞24 h; CsA+TAM组,经200 ng/mL CsA处理肿瘤相关巨噬细胞的培养液上清干预Ishikawa细胞24 h。处理结束后,CCK-8法检测细胞活性,Transwell小室实验检测细胞迁移与侵袭,Annexin V-FITC/PI双染法检测细胞凋亡,Western Blotting检测自噬相关蛋白表达,免疫荧光染色检测自噬标记物LC3表达,透射电子显微镜观察自噬体形成。结果:经过不同浓度CsA处理肿瘤相关巨噬细胞后,细胞存活率未发生变化(P>0.05),而细胞中CD86表达增加、CD206表达下降,iNOS、TNF-α的mRNA相对表达量升高,TGF-β、Arg-1的mRNA相对表达量降低(P<0.01);与对照组比较,CsA组Ishikawa细胞存活率下降,迁移数目与侵袭数目减少,细胞凋亡率增加,Beclin-1蛋白表达下降、p62蛋白表达增加,LC3-Ⅱ/LC3-Ⅰ比值下调,LC3荧光表达减弱,细胞内未见自噬体,而TAM组细胞迁移数目与侵袭数目增加,细胞凋亡率减少,Beclin-1蛋白表达增加、p62蛋白表达下降,LC3-Ⅱ/LC3-Ⅰ比值上调,LC3荧光表达增强,细胞内自噬体数目较多(P<0.01);与TAM组比较,CsA+TAM组细胞存活率下降,迁移数目与侵袭数目减少,细胞凋亡率增加,Beclin-1蛋白表达下降、p62蛋白表达增加,LC3-Ⅱ/LC3-Ⅰ比值下调,同时,LC3荧光表达减弱,细胞内自噬体减少(P<0.01)。结论:CsA通过调控肿瘤相关巨噬细胞的极化,抑制子宫内膜癌细胞增殖、迁移与侵袭能力,其机制可能与细胞自噬水平相关。
引用
收藏
页码:2690 / 2698
页数:9
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