ALDH1A1增强乳腺癌细胞血管生成因子表达并促进共培养HUVEC细胞小管形成和侵袭能力  

ALDH1A1 enhances the expression of angiogenesis factor in breast cancer cells and promotes tubule formation and invasion of co-cultured HUVEC cells

作　　者：赵洪玉[1] 阿不都艾尼·图尔逊 范明江 ZHAO Hongyu;Abduhini·Tursun;FAN Mingjiang(Surgical Oncology of Breast and Thyroid,the First People's Hospital of Kashgar,Xinjiang Kashgar 844000,China)

机构地区：[1]喀什地区第一人民医院乳腺甲状腺肿瘤外科,新疆喀什844000

出　　处：《现代肿瘤医学》2021年第4期552-559,共8页Journal of Modern Oncology

基　　金：新疆维吾尔自治区科技支疆项目(编号:2017E0263)。

摘　　要：目的:探讨干细胞标志物醛脱氢酶1A1(ALDH1A1)对乳腺癌细胞血管生成因子表达的影响,以及对与乳腺癌细胞共培养的HUVEC细胞小管形成和侵袭能力的影响。方法:采用免疫组化检测了乳腺癌组织和乳腺增生组织中ALDH1A1的表达。使用ALDH1A1 shRNA或过表达ALDH1A1的pcDNA3.1质粒转染乳腺癌细胞(MCF-7和MDA-MB-231),通过qRT-PCR和Western blot检测敲低或过表达ALDH1A1对乳腺癌细胞中血管内皮生长因子(VEGF)、缺氧诱导因子-1α(HIF-1α)和白细胞介素-12(IL-12)表达的影响。通过用1μmol/L的外源性RA和RAR阻断剂(AGN 193109)处理乳腺癌细胞48 h来考察视黄酸信号通路是否参与ALDH1A1对VEGF和HIF-1α的调控过程。将乳腺癌细胞(MCF-7和MDA-MB-231)和HUVEC细胞共培养来模拟肿瘤形成的微环境,并检测HUVEC的小管形成能力和细胞侵袭能力。结果:乳腺癌组织的ALDH1A1染色平均光密度显著高于乳腺增生组织,并且淋巴结转移的乳腺癌组织显著高于未淋巴结转移的乳腺癌组织(P<0.05)。敲低ALDH1A1可显著降低MCF-7和MDA-MB-231细胞中VEGF和HIF-1α蛋白表达,并上调IL-12蛋白表达。然而,上调ALDH1A1表达则可逆转上述变化。外源性RA处理可显著上调MCF-7和MDA-MB-231细胞中VEGF和HIF-1α的表达,然而,RAR阻断剂处理可抑制MCF-7和MDA-MB-231细胞中VEGF和HIF-1α的上调。敲低乳腺癌细胞中ALDH1A1的表达可导致共培养的HUVEC细胞的小管形成能力和侵袭能力显著降低。而上调乳腺癌细胞中ALDH1A1的表达则可显著促进共培养的HUVEC细胞的小管形成能力和侵袭能力。结论:在乳腺癌细胞中,ALDH1A1通过激活HIF-1α和视黄酸信号通路来上调血管生成因子的表达并提高共培养的内皮细胞的血管生成能力,从而增加肿瘤的侵袭性。Objective:To investigate the effect of stem cell marker aldehyde dehydrogenase 1A1(ALDH1A1)on the expression of angiogenic factors in breast cancer cells and the effect of tubule formation and invasion on HUVEC cells co-cultured with breast cancer cells.Methods:The expression of ALDH1A1 in breast cancer tissues and breast hyperplasia tissues was detected by immunohistochemistry.Breast cancer cells(MCF-7 and MDA-MB-231)were transfected with ALDH1A1 shRNA or overexpressing ALDH1A1-pcDNA3.1 plasmid,and the effects of knockdown or overexpression of ALDH1A1 on the expression of vascular endothelial growth factor(VEGF),hypoxia inducible factor-1α(HIF-1α)and interleukin-12(IL-12)in breast cancer cells were detected by qRT-PCR and Western blot.Breast cancer cells were treated with exogenous RA and RAR blocker(AGN 193109)for 48 hours to investigate whether retinoic acid signaling pathway was involved in the regulation of ALDH1A1 on VEGF and HIF-1α.Breast cancer cells(MCF-7 and MDA-MB-231)and HUVEC cells were co-cultured to mimic the microenvironment of tumor formation,and the tubular formation ability and cell invasion ability of HUVEC were examined.Results:The mean optical density of ALDH1A1 staining in breast cancer tissues was significantly higher than that in breast hyperplasia tissues,and the lymph node metastasis of breast cancer tissues was significantly higher than that in non-lymph node metastatic breast cancer tissues(P<0.05).Knockdown of ALDH1A1 significantly reduced VEGF and HIF-1αprotein expression and up-regulated IL-12 protein expression in MCF-7 and MDA-MB-231 cells.However,up-regulation of ALDH1A1 expression reversed these changes.Exogenous RA treatment significantly up-regulated the expression of VEGF and HIF-1αin MCF-7 and MDA-MB-231 cells,however,RAR blocker treatment inhibited the up-regulation of VEGF and HIF-1αin MCF-7 and MDA-MB-231 cells.Knockdown of ALDH1A1 expression in breast cancer cells resulted in a significant decrease in tubule formation and invasive ability of co-cultured HUVEC cell

关 键 词：乳腺癌 醛脱氢酶1A1 视黄酸 缺氧诱导因子-1Α 侵袭 血管生成 

分 类 号：R737.9[医药卫生—肿瘤]