Suppression of Spry4 enhances cancer stem cell properties of human MDA-MB-231 breast carcinoma cells.
Suppression of Spry4 enhances cancer stem cell properties of human MDA-MB-231 breast carcinoma cells.
复制标题
DOI：
10.1186/s12935-016-0292-7
复制
发表时间：
2016
影响因子：
5.8
通讯作者：
Yang X
中科院分区：
文献类型：
--
作者：
Jing H;Liaw L;Friesel R;Vary C;Hua S;Yang X
癌症干细胞有助于肿瘤的发生、异质性和复发，并且是癌症治疗中的关键靶点。Sprouty 4（Spry 4）是一种有效的酪氨酸激酶受体信号转导抑制剂，在调节细胞增殖、迁移和分化中发挥重要作用。Spry 4被认为是一种肿瘤抑制因子，并参与调节胚胎干细胞。这项研究的目的是测试Spry 4调节乳腺癌中癌症干细胞特性的新想法。利用Spry 4在人MDA-MB-231细胞中的功能缺失来验证我们的假设。使用人Spry 4或非靶向对照shRNA的慢病毒递送产生Spry 4敲低或对照细胞系，然后用2 μg/ml嘌呤霉素选择。细胞生长和迁移能力分别用生长曲线和细胞周期流式细胞仪分析和划痕试验测定。采用异种移植瘤模型测定其体内致瘤活性和转移情况。使用免疫印迹分析和荧光激活细胞分选来评估癌症干细胞相关标志物。使用体外乳腺球形成和药物敏感性测试以及体内有限稀释肿瘤形成测定来评估癌症干细胞表型。三种测试的人Spry 4 shRNA中的两种显著抑制MDA-MB-231细胞中内源性Spry 4的表达。抑制Spry 4表达增加MDA-MB-231细胞增殖和迁移。抑制Spry 4增加β3整合素表达和CD 133 + CD 44+亚群。抑制Spry 4增加了乳腺球的形成，同时降低了MDA-MB-231细胞对紫杉醇治疗的敏感性。最后，抑制Spry 4增加了MDA-MB-231细胞肿瘤起始的效力，这是归因于癌症干细胞的特征。我们的研究结果提供了新的证据，即内源性Spry 4可能通过抑制癌症干细胞特性以及对肿瘤细胞增殖和迁移的负面影响而在乳腺癌中具有肿瘤抑制活性。本文的在线版本（doi：10.1186/s12935-016-0292-7）包含补充材料，可供授权用户使用。
Cancer stem cells contribute to tumor initiation, heterogeneity, and recurrence, and are critical targets in cancer therapy. Sprouty4 (Spry4) is a potent inhibitor of signal transduction pathways elicited by receptor tyrosine kinases, and has roles in regulating cell proliferation, migration and differentiation. Spry4 has been implicated as a tumor suppressor and in modulating embryonic stem cells. The purpose of this research was to test the novel idea that Spry4 regulates cancer stem cell properties in breast cancer. Loss-of function of Spry4 in human MDA-MB-231 cell was used to test our hypothesis. Spry4 knockdown or control cell lines were generated using lentiviral delivery of human Spry4 or non-targeting control shRNAs, and then selected with 2 μg/ml puromycin. Cell growth and migratory abilities were determined using growth curve and cell cycle flow cytometry analyses and scratch assays, respectively. Xenograft tumor model was used to determine the tumorigenic activity and metastasis in vivo. Cancer stem cell related markers were evaluated using immunoblotting assays and fluorescence-activated cell sorting. Cancer stem cell phenotype was evaluated using in vitro mammosphere formation and drug sensitivity tests, and in vivo limiting dilution tumor formation assay. Two out of three tested human Spry4 shRNAs significantly suppressed the expression of endogenous Spry4 in MDA-MB-231 cells. Suppressing Spry4 expression increased MDA-MB-231 cell proliferation and migration. Suppressing Spry4 increased β3-integrin expression, and CD133+CD44+ subpopulation. Suppressing Spry4 increased mammosphere formation, while decreasing the sensitivity of MDA-MB-231 cells to Paclitaxel treatment. Finally, suppressing Spry4 increased the potency of MDA-MB-231 cell tumor initiation, a feature attributed to cancer stem cells. Our findings provide novel evidence that endogenous Spry4 may have tumor suppressive activity in breast cancer by suppressing cancer stem cell properties in addition to negative effects on tumor cell proliferation and migration. The online version of this article (doi:10.1186/s12935-016-0292-7) contains supplementary material, which is available to authorized users.