Silencing of HSulf-2 expression in MCF10DCIS.com cells attenuate ductal carcinoma in situ progression to invasive ductal carcinoma in vivo.
Silencing of HSulf-2 expression in MCF10DCIS.com cells attenuate ductal carcinoma in situ progression to invasive ductal carcinoma in vivo.
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DOI：
10.1186/bcr3140
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发表时间：
2012-03-12
期刊：
影响因子：
--
通讯作者：
Shridhar V
中科院分区：
文献类型：
--
作者：
Khurana A;McKean H;Kim H;Kim SH;mcguire J;Roberts LR;Goetz MP;Shridhar V
乳腺导管原位癌（DCIS）是一组异质性增生细胞病变，有可能成为浸润性的。关于从DCIS进展为浸润性导管癌（IDC）的分子改变知之甚少。乙酰肝素内切硫酸酯酶（HSulf-2）编辑硫酸乙酰肝素蛋白聚糖（HSPG）上的硫酸酯部分，并且涉及调节肝素结合生长因子信号传导、血管生成和肿瘤发生。然而，HSulf-2在乳腺癌进展中的作用知之甚少。MCF10DCIS.com细胞（称为MCF 10 DCIS）表达HSulf-2并形成粉刺型DCIS，并且当移植到免疫缺陷小鼠中时进展为IDC，因此，是研究乳腺癌进展的理想模型。我们使用小鼠脂肪垫乳腺异种移植物评估了HSulf-2在从DCIS进展到IDC中的作用。分别用NTC shRNA和两种不同的针对HSulf-2的慢病毒shRNA在MCF 10 DCIS乳腺癌细胞中实现了非靶向对照（NTC）和HSulf-2的敲低。通过在小鼠乳腺脂肪垫中注射NTC和HSulf-2缺陷型MCF 10 DCIS细胞建立异种移植物。对异种移植物进行H&E染色以进行形态学分析、TUNEL和碘化丙啶染色（以确定细胞凋亡的程度）、蛋白质印迹分析和酶谱分析。使用小鼠乳房脂肪垫衍生的异种移植物模型，我们观察到与对照处理的异种移植物相比，HSulf-2的下调与第7周的生长显著延迟相关（P值< 0.05）。肿瘤的组织学检查表明，在HSulf-2耗尽的异种移植物中，粉刺状坏死具有显著的管腔凋亡和凋亡标志物Bim、裂解的PARP和裂解的半胱天冬酶3的上调。此外，与非靶向对照相比，HSulf-2耗尽的异种移植物保留了基底膜的完整性，基质金属蛋白酶9（MMP-9）的活性和表达降低，MMP-9是细胞外基质降解的关键酶。我们的数据表明，HSulf-2表达可能是人类乳腺癌进展的关键。HSulf-2的下调导致粉刺型DCIS的保留并延迟DCIS向IDC的进展。有必要进一步研究以确定HSulf-2表达的治疗靶向是否可能延迟DCIS向IDC的进展。
Ductal carcinoma in situ (DCIS) of the breast is a heterogeneous group of proliferative cellular lesions that have the potential to become invasive. Very little is known about the molecular alterations involved in the progression from DCIS to invasive ductal carcinoma (IDC). Heparan endosulfatase (HSulf-2) edits sulfate moieties on heparan sulfate proteoglycans (HSPGs) and has been implicated in modulating heparin binding growth factor signaling, angiogenesis and tumorigenesis. However, the role of HSulf-2 in breast cancer progression is poorly understood. MCF10DCIS.com cells (referred as MCF10DCIS) express HSulf-2 and form comedo type DCIS and progress to IDC when transplanted in immune-deficient mice and, therefore, is an ideal model to study breast cancer progression. We evaluated the role of HSulf-2 in progression from DCIS to IDC using mouse fat pad mammary xenografts. Non-target control (NTC) and HSulf-2 knockdown in MCF10DCIS breast cancer cells were achieved by NTC shRNA and two different lentiviral shRNA against HSulf-2 respectively. Xenografts were established by injecting NTC and HSulf-2 deficient MCF10DCIS cells in mouse mammary fat pads. Xenografts were subjected to H&E staining for morphological analysis, TUNEL and Propidium iodide staining (to determine the extent of apoptosis), Western blot analysis and zymography. Using a mouse mammary fat pad derived xenograft model, we observed that compared to control treated xenografts, down-regulation of HSulf-2 was associated with significant delays in growth at Week 7 (P-value < 0.05). Histological examination of the tumors demonstrated substantial differences in comedo necrosis, with marked luminal apoptosis and up-regulation of apoptotic markers Bim, cleaved PARP and cleaved caspase 3 in HSulf-2 depleted xenografts. Furthermore, HSulf-2 depleted xenografts retained the basement membrane integrity with decreased activity and expression of matrix metalloproteinase 9 (MMP-9), an enzyme critical for degradation of extracellular matrix compared to nontargeted control. Our data suggest that HSulf-2 expression may be critical for human breast cancer progression. Down-regulation of HSulf-2 leads to retention of comedo type DCIS and delays the progression of DCIS to IDC. Further studies are necessary to determine if therapeutic targeting of HSulf-2 expression might delay the progression of DCIS to IDC.
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DOI：
10.1111/j.1478-3231.2010.02336.x
发表时间：
2010-11
期刊：
Liver international : official journal of the International Association for the Study of the Liver
影响因子：
--
作者：
Lai JP;Sandhu DS;Yu C;Moser CD;Hu C;Shire AM;Aderca I;Murphy LM;Adjei AA;Sanderson S;Roberts LR
通讯作者：
Roberts LR
影响因子：
3.7
作者：
Nawroth R;van Zante A;Cervantes S;McManus M;Hebrok M;Rosen SD
通讯作者：
Rosen SD
影响因子：
4.8
作者：
Locatelli, Alessia;Lange, Carol A.
通讯作者：
Lange, Carol A.
影响因子：
7.4
作者：
Brown, A M
通讯作者：
Brown, A M
影响因子：
8
作者：
Lemjabbar-Alaoui, H.;van Zante, A.;Singer, M. S.;Xue, Q.;Wang, Y-Q;Tsay, D.;He, B.;Jablons, D. M.;Rosen, S. D.
通讯作者：
Rosen, S. D.