蓝萼乙素对三阴性乳腺癌 MDA-MB-231细胞增殖、迁移及侵袭的影响

目的:探讨蓝萼乙素对三阴性乳腺癌MDA-MB-231细胞增殖、迁移及侵袭的影响及其作用机制。方法:体外培养人乳腺癌细胞MDA-MB-231,加入不同浓度(0、2、4、8μmol/L)蓝萼乙素干预处理,采用MTT法检测细胞增殖能力;划痕实验考察细胞迁移能力;Transwell小室法考察细胞侵袭能力;Western blotting法检测细胞内p38MAPK、p-p38MAPK、FOXO3a及上皮-间质转化(EMT)相关标志物(E-cadherin、Vimentin及N-cadherin)的表达水平。结果:蓝萼乙素浓度为2、4、8μmol/L时,细胞增殖率分别为(83.2±6.90)%、(70.72±6.53)%、(45.43±7.51)%,较空白对照组[(100.00±7.84)%]降低;细胞侵袭数量分别为(300.54±24.91)、(255.44±23.59)、(208.66±36.18),较空白对照组(368.44±28.32)降低;细胞迁移距离分别为(487.11±53.00)μm、(394.93±61.91)μm、(312.88±35.42)μm,较空白对照组[(559.37±75.77)μm]降低;p-p38MAPK表达量分别为(1.38±0.11)、(1.69±0.13)、(2.23±0.19),较空白对照组(1.00±0.09)增加;FOXO3α表达量分别为(1.40±0.16)、(1.97±0.31)、(2.44±0.26),较空白对照组(1.00±0.18)增加;E-cadherin表达量分别为(1.15±0.11)、(1.77±0.22)、(1.86±0.15),较空白对照组(1.00±0.11)增加;Vimentin表达量分别为(0.86±0.04)、(0.49±0.05)、(0.54±0.04),较空白对照组(1.00±0.04)减少;N-cadherin表达量分别为(0.66±0.07)、(0.58±0.08)、(0.42±0.04),较空白对照组(1.00±0.12)减少,差异均有统计学意义(P 0.05)。结论:蓝萼乙素可通过介导p38MAPK/FOXO3a信号传导有效干扰肿瘤细胞EMT进程,发挥其抑制三阴性乳腺癌细胞增殖、迁移及侵袭的作用。     

川楝素对乳腺癌MDA-MB-231细胞增殖的抑制作用

目的 探讨川楝素对人三阴性乳腺癌MDA-MB-231细胞生长的抑制作用及其机制.方法 取对数生长期的MDA-MB-231细胞,采用0、6.25、12.50、25.00、50.00和100.00 nmol/L川楝素处理,绘制生长曲线.川楝素0、12.5和50 nmol/L处理72 h后,采用MTS法检测川楝素对MDA-MB-231细胞增殖,流式细胞仪检测乳腺癌细胞凋亡和周期,Western blot检测凋亡相关蛋白半胱氨酶蛋白酶3(Caspase-3)、聚腺苷酸二磷酸核糖转移酶(PARP)和cleave-PARP表达.结果 川楝素呈时间和剂量依赖性抑制MDA-MB-231乳腺癌细胞增殖(P＜0.01).川楝素作用48、72和96 h的半数抑制浓度(IG0)分别为9.32、16.96和122.37nmol/L.川楝素能呈浓度依赖性诱导乳腺癌细胞凋亡(P＜0.01),阻滞细胞在S期(P＜0.01).以0、12.50和50.00 nmol/L川楝素处理MDA-MB-231细胞72 h,其早期凋亡率分别为8.12％、20.85％和67.21％,处于细胞周期S期的细胞占32.69％、47.90％和61.23％,凋亡相关蛋白Caspase-3和PARP减少,cleaved-PARP增多.结论 川楝素对人乳腺癌MDA-MB-231细胞增殖具有抑制作用;其机制可能与诱导细胞凋亡和引起细胞S期阻滞有关.     

Triptolide inhibits human breast cancer MCF-7 cell growth via downregulation of the ERα-mediated signaling pathway

Aim:

To investigate the anticancer mechanisms of triptolide, a diterpenoid isolated from the plant Tripterygium wilfordii Hook F, against human breast cancer cells and the involvement of the estrogen receptor-α (ERα)-mediated signaling pathway in particular.

Methods:

Human breast cancer ERα-positive MCF-7 cells and ERα-negative MDA-MB-231 cells were tested. PrestoBlue assay was used to evaluate the cell viability. The levels of ERα mRNA and protein were detected with real-time PCR and immunoblotting, respectively. Mouse models of MCF-7 or MDA-MB-231 xenograft tumors were treated with triptolide (0.4 mg·kg⁻¹·d⁻¹, po) or a selective estrogen receptor modulator tamoxifen (mg·kg⁻¹·d⁻¹, po) for 3 weeks, and the tumor weight and volume were measured.

Results:

Triptolide (5–200 nmol/L) dose-dependently inhibited the viability of both MCF-7 and MDA-MB-231 cells, with a more potent inhibition on MCF-7 cells. Knockdown of ERα in MCF-7 cells by siRNA significantly attenuated the cytotoxicity of triptolide, whereas overexpression of ERα in MDA-MB-231 cells markedly enhanced the cytotoxicity. Triptolide dose-dependently decreased the expression of ERα in MCF-7 cells and MCF-7 xenograft tumors. Furthermore, treatment of MCF-7 cells with triptolide inhibited the phosphorylation of ERK1/2 in dose- and time-dependent manners. In the mice xenografted with MCF-7 cells, treatment with triptolide or tamoxifen resulted in significant reduction in the tumor weight and volume. Similar effects were not obtained in the mice xenografted with MDA-MB-231 cells.

Conclusion:

The anticancer activity of triptolide against ERα-positive human breast cancer is partially mediated by downregulation of the ERα-mediated signaling pathway.     

circRNA_001569通过miR-145/HBXIP轴影响乳腺癌细胞的恶性生物学行为

目的：探讨 circRNA_001569 通过 miR-145/HBXIP 轴在乳腺癌细胞增殖、侵袭、迁移中发挥的作用。方法:收集2016年1月至2019年1月期间衡水市人民医院收治的30例乳腺癌患者的癌组织和癌旁组织。qPCR检测circRNA_001569在乳腺癌组织、癌旁组织以及细胞系中的表达。生物信息学工具预测miR-145的靶基因，RNA免疫沉淀（RNA immunoprecipitation，RIP）和双荧光素酶报告基因实验检测 miR-145 或靶基因之间的相互作用 ；向乳 腺 癌 MDA-MB-231 和 MCF-7细胞中转染si-circRNA_001569、miR-145 mimics或miR-145 inhibitor，建立基因过表达或沉默的细胞模型，qPCR和Western blotting分别检测转染对相关基因和蛋白表达的影响，CCK-8法、Transwell实验检测转染对细胞增殖、侵袭和迁移的影响。结果:在乳腺癌组织和乳腺癌细胞中，circRNA_001569 和 HBXIP 均呈高表达、miR-145 呈低表达。RIP 分析和双荧光素酶实验证实了 miR-145 与circRNA_001569和HBXIP之间的靶向关系；circRNA_001569或HBXIP过表达促进MDA-MB-231和MCF-7细胞的增殖、侵袭和迁移（均 P<0.01），而 miR-145 过表达起相反的作用（均 P<0.01）。结论:circRNA_001569 可能通过下调 miR-145 的表达、上调HBXIP的表达从而促进乳腺癌细胞的增殖、侵袭和迁移。     

2-Methoxy-1,4-Naphthoquinone (MNQ) suppresses the invasion and migration of a human metastatic breast cancer cell line (MDA-MB-231)

Metastasis contributes to the escalating mortality rate among cancer patients worldwide. The search for novel and more effective anti-metastatic agent is crucial owing to the lack of anticancer drugs that can successfully combat metastasis. Hence, this study aims to examine the effects of 2-Methoxy-1,4-Naphthoquinone (MNQ) towards the metastasis of MDA-MB-231 cells. In invasion assays, the number of cells permeating across a Matrigel barrier was found to be decreased in a dose-dependent manner upon treatment with MNQ (0–7.5 μM). In wound-healing migration assays, MNQ exhibited dose-dependent inhibition of cell migration in which significant reduction in the zone of closure was observed as compared to untreated controls. Furthermore, the proteolytic activity of a pivotal metastatic mediator, matrix metalloproteinase-9 (MMP-9) was also downregulated by MNQ as determined by gelatin zymography. This study reports for the first time, the ability of MNQ to inhibit the invasion and migration characteristics of a highly metastatic MDA-MB-231 cancer cell line.     

Novel Suppressive Effects of Ketotifen on Migration and Invasion of MDA-MB-231 and HT-1080 Cancer Cells

The high mortality rates associated with cancer reflect the metastatic spread of tumor cells from the site of their origin. Metastasis, in fact, is the cause of 90% of cancer deaths. Therefore, considerable effort is being made to inhibit metastasis. In the present study, we screened ketotifen for anti-migratory and anti-invasive activities against MDA-MB-231 breast cancer and HT-1080 fibrosarcoma cancer cells. Cancer cell migration and invasion were measured using multi-well chambers. Additionally, western blots were used to examine the effects of ketotifen on the expressions of CDC42, Rho, Rac, and matrix metalloproteinase 9 (MMP-9). The results showed that ketotifen dose-dependently suppressed the migration and invasion of MDA-MB-231 and HT-1080 cells. Ketotifen also suppressed the expressions of CDC42, Rac, and Rho, which, significantly, are involved in MDA-MB-231 and HT-1080 cancer cell migration. Moreover, ketotifen suppressed the expression and activity of MMP-9, which is involved in degradation of the extracellular matrix leading to invasion. The overall data suggested that ketotifen suppresses the migration and invasion of MDA-MB-231 and HT-1080 cancer cells via inhibition of CDC42, Rac, Rho, and MMP-9 expression.     

Effect of colestipol and clofibrate on plasma lipid and lipoproteins in Type IIa hyperlipoproteinemia

The effects of 2.0 g of clofibrate and 15, 20 and 30 g of colestipol on plasma lipid and lipoprotein levels were evaluated in adult patients with Type IIa hyperlipoproteinemia. Clofibrate treatment was associated with decreases of 11.0% in plasma cholesterol, 15.2% in LDL cholesterol, 26.1% in triglycerides, and an 11.3% increase in HDL cholesterol. The reductions in total cholesterol with the various doses of colestipol ranged from 11.9 to 17.8% and reductions in LDL cholesterol ranged from 16.1 to 27.3%. Colestipol treatment was not associated with any significant change in HDL cholesterol levels and minor increases in triglycerides. The addition of clofibrate to patients receiving colestipol resulted in a significant increase in HDL cholesterol and a decrease in triglycerides, but no additional reduction in total or LDL cholesterol.     

25-Methoxyhispidol A, a novel triterpenoid of Poncirus trifoliata, inhibits cell growth via the modulation of EGFR/c-Src signaling pathway in MDA-MB-231 human breast cancer cells

The fruit of Poncirus trifoliata (Rutaceae) has been used a medicinal food and traditional medicine. Recently we reported the isolation of 25-methoxyhispidol A (25-MHA) as a novel triterpenoid from the immature fruit of P. trifoliata with the potential growth inhibition of cancer cells. However, the molecular mechanisms on the anti-proliferative activity in cancer cells remain to be elucidated. In the present study, we investigated the anti-proliferative activity and mechanisms of actions mediated by 25-MHA in estrogen receptor (ER)-negative MDA-MB-231 human breast cancer cells. 25-MHA exhibited the growth inhibitory activity against MDA-MB-231 cells with the cell cycle arrest in the G0/G1 phase. The cell cycle arrest in the G0/G1 by 25-MHA was well correlated with the downregulation of cyclin D1, cyclin dependent kinase (CDK4), CDK2, cyclin A, phosphorylated retinoblastoma protein (pRb), and induction of cdk inhibitor p21^WAF1/Cip1 protein. 25-MHA also suppressed the activation of c-Src/epidermal growth factor receptor (EGFR)/Akt signaling, and consequently led to the inactivation of mTOR and its downstream signal molecules including 4E-binding protein (4E-BP) and p70 S6 kinase. These findings suggest that 25-MHA-mediated inhibitory activity of human breast cancer cell growth might be related with the cell cycle arrest and modulation of signal transduction pathways.     

红景天提取物对乳腺癌、宫颈癌治疗的研究综述

体内外的肿瘤研究显示红景天具有一定的抗肿瘤作用,本文综述了红景天提取物对乳腺癌MDA-MB-435细胞株及宫颈癌Hela细胞的影响及其可能的机制。显示：红景天治疗后移植瘤内乳腺癌细胞增殖指标Ki-67染色比例、染色强度降低,增殖指标PCNA的染色强度和比例的综合评价指数H分数均值有所降低;红景天治疗后宫颈癌Hela细胞胞体回缩,贴附型细胞不贴壁,胞质粗糙,有大量颗粒状物堆积,而且药物浓度越大,形态学改变越明显。给药组肿瘤细胞增殖缓慢甚至停滞,出现细胞脱落、胞浆内颗粒状物堆积等形态学改变,克隆形成数明显少于对照组,cpm和A值明显降低,即3H-TdR掺入率减少,生存率下降。结果表明红景天的体内抗癌机制可能部分通过抑制肿瘤的增殖。