目的:分析核仁纺锤体相关蛋白(NuSAP1)的表达对乳腺癌(BC)患者预后的影响,探讨三阴性乳腺癌(TNBC)细胞株的致瘤机制.方法:我们从癌症基因组图谱(TCGA)下载了乳腺癌(BC)的RNA-seq数据,用R软件在此数据中筛选出了NuSAP1基因及BC患者的临床资料.生存曲线用Kaplan-Meier-plotter绘制.建立阴性对照组(NC)、敲减NuSAP1组(sh-NuSAP1)、过表达NuSAP1组(sh-NuSAP1+ov-NuSAP1),用细胞增殖毒性检测试剂盒(CCK8)和Transwell法检测各组细胞增殖和侵袭能力.实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,qRT-PCR)和免疫印迹法(Western blotting)检测肿瘤细胞中NuSAP1、Wnt/β-catenin通路和上皮-间充质转化(epithelial-mesenchymal transition,EMT)相关蛋白的表达.结果:NuSAP1在BC中表达上调.NuSAP1表达差异与多种临床病理因素有关,且NuSAP1表达越高,生存期越短.在MDA-MB-231和BT-549细胞中,与NC组相比,sh-NuSAP1组细胞增殖和侵袭能力下降,CyclinD1、Vimentin、Slug、Twist、Wnt3a和p-β-catenin的表达减少,而E-cadherin的表达显著增加.与sh-NuSAP1组相比,sh-NuSAP1+ov-NuSAP1组结果与上述相反.结论:NuSAP1是一种致癌基因,预测乳腺癌的不良预后,并通过Wnt/β-catenin和上皮-间充质转化(EMT)途径促进肿瘤进展.

Breast cancer (BC) is a leading cause of cancer mortality in women worldwide. MAC30/Transmembrane protein 97 (TMEM97) is aberrantly up-regulated in many human carcinoma cells. However, the function of MAC30 in invasion and EMT of BC cells is uncertain. qRT-PCR was used to determine the level of MAC30 in BC tissues and cell lines. si-MAC30 was transfected into BC cells, and the effects of MAC30 silencing on the invasion and EMT were explored by qRT-PCR as well as transwell and western blot assays. Also, we determined the effects of MAC30 silencing on Wnt/β-catenin and PI3K/Akt signaling pathways by western blot. We found that MAC30 is significantly up-regulated in BC tissues and cell lines. Down-regulation of MAC30 expression efficiently inhibited the invasion of BC cells. Furthermore, the EMT of BC cells was also inhibited by down-regulation of MAC30. Finally, we found that MAC30 knockdown inhibited Akt phosphorylation, β-catenin, survivin, and cyclin D1 expressions. To our knowledge, this is the first report investigating the effect of MAC30 on invasion and EMT in BC cells by suppressing Wnt/β-catenin and PI3K/Akt signaling pathways. MAC30 may be a potential therapeutic target for BC.

目的 探讨miR-1468-3p分子通过Janus激酶2(JAK2)蛋白调控乳腺癌细胞的凋亡.方法 运用TargetScan在线分析miR-1468-3p与JAK2的相关性;将JAK2的YUTR构建进PmirGLO质粒,利用luciferase assay 检测miR-1468-3p是否靶向调控JAK2;用脂质体梯度转染miR-1468-3P mimics或miR-1468-3P inhibitor转入乳腺癌MCF7细胞,通过Western blot检测JAK2的表达量和乳腺癌MCF7细胞凋亡标志蛋白表达情况.结果 通过TargetScan分析,miR-1468-3p在3个区域与JAK2具有较高匹配度;通过luciferase assay发现,miR-1468-3p靶向JAK2的YUTR;梯度转染miR-1468-3P mimics时,JAK2的表达量梯度下降,细胞凋亡标志蛋白cleaved-caspase3/caspase3、cleaved-caspase9/caspase9、BAX表达量上升,Bcl-2表达量下降(P＜0.05);而用梯度转染miR-1468-3P inhibitor 进乳腺癌MCF7细胞时,JAK2的表达量梯度上升,cleaved-caspase3/caspase3、cleaved-caspase9/caspase9和BAX表达量下降,Bcl-2表达量上升(P＜0.05).结论 miR-1468-3p能通过降低JAK2的表达来促进乳腺癌MCF7细胞的凋亡.

Down-regulation of the meningioma-associated protein (MAC30) gene has been found in many solid cancers. This study was carried out to determine the roles and the mechanisms of MAC30 in breast cancer. We used our own data and a public database to analyze the MAC30 mRNA and protein levels in breast cancer tissues. In addition, we established MAC30 knockdown breast cancer cells using MAC30 siRNA. The roles of MAC30 were detected by using the Soft agar assay, Annexin-V-FITC/PI double staining and the Transwell assay. Western blotting was used to analyze the potential mechanism(s) of MAC30 in these cells. We found that MAC30 mRNA and protein were higher in the cancer tissues compared to the matched normal tissues. MAC30 expression was associated with tumor size, tumor differentiation and estrogen receptor (ER) status. Overall survival rate of the patients with low MAC30 expression was obviously higher than the ones with high expression. The apoptotic ratio was lower in MDA-MB-231 and MDA-MB-157 cells with MAC30 expression. By Western blot analysis, we found that increased levels of phosphorylated YAP1, MST1 and LATS1 after MAC30 siRNA transfection in these two cells. In summary, we demonstrate that MAC30 knockdown is involved in the activation of the Hippo signaling pathway.

Breast cancer is the most frequently diagnosed cancer in females. Warburg effect could enhance tumorigenesis and has garnered attention as a target for tumor treatment. In this study, we found that the mRNA and protein levels of hexokinase 2 (HK2), pyruvate kinase (PKM2), and pyruvate dehydrogenase kinase (PDK1) in breast cancer tissues were higher than those in corresponding noncancerous tissues. HK2, PKM2, and PDK1 expression was correlated statistically with the survival rate of the patients with breast cancer. We also demonstrated a shorter fragment of KISS1, Kisspeptin-10 (KP-10), inhibited the Warburg effect and induced mitochondrial injury in human breast cancer cell line, MDA-MB-231. We confirmed that KP-10-inhibited the Warburg effect by activating Smad pathway. The effects and related mechanisms of these treatments were also confirmed in murine xenografts. However, additional studies are needed to confirm these results in other cell types.

Breast cancer is the leading cause of cancer-related death in women. Kisspeptin-10 (KP-10) is a shorter fragment of KISS1. In the present study, we demonstrated the antitumor effects of KP-10 on human breast cancer cell lines, MDA-MB-231 and MDA-MB-157, both in vitro and in vivo. KP-10 was observed to induce apoptosis and inhibit the mobility of MDA-MB-231 and MDA-MB-157 cells. Correspondingly, KP-10 suppressed tumor growth in established xenograft tumor models and improved the survival rate of tumor-bearing mice. The formation of intratumoral microvessels was inhibited following treatment with KP-10. Finally, we confirmed that KP-10 inhibited cell mobility via epithelial-mesenchymal transition (EMT). Overall, the present study demonstrated that KP-10 suppressed breast cancer and human umbilical vein endothelial cell (HUVEC) growth both in vivo and in vitro. KP-10 is a novel regulator of EMT in breast cancer cells. However, additional studies are needed to confirm these results in other cell types.

Breast cancer is the most frequently diagnosed cancer in women. In these studies, a metastasis suppressor gene, KISS1 and its truncated fragment, were overexpressed in the breast cancer cell line MDA-MB-231. In addition, KISS1 expression was downregulated in MDA-MB-157 cell line using a KISS1-specific siRNA. The effects of KISS1 on breast cancer cells both in vivo and in vitro were then identified. Our results indicate that KISS1 can induce apoptosis and inhibit mobility of breast cancer cells. Moreover, the expression of KISS1 in established xenografted tumors was associated with a decrease in tumor size and weight. Accordingly, the survival rate of these mice was significantly higher compared to that of mice bearing tumors that did not express KISS1. We also confirm that KISS1 could decrease the number of circulating tumor cells (CTCs). The plasma levels of metastin and the number of CTCs were significantly positively correlated. Furthermore, we found that KISS1 can inactivate p-MEK and p-ERK. Overall, these studies demonstrate the antitumor activity of KISS1 in the breast cancer cell lines and provide insight into relevant mechanisms that may lead to novel treatments for breast cancer.