Possible mechanism of 191 measles attenuated live vaccine strains inhibiting the proliferation of triple negative breast cancer MDA-MB-231 cells based on proteomics technology / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：基于蛋白质组学技术探讨麻疹减毒活疫苗191株（MV-Hu191）在体内外对三阴性乳腺癌MDA-MB-231、4T1细胞的影响及其作用机制。方法：采用CCK-8法检测MV-Hu191对MDA-MB-231和4T1细胞增殖的影响；液相色谱-质谱联用技术分析MV-Hu191处理对MDA-MB-231细胞中蛋白质谱的影响，多重数据库筛选蛋白质谱中的典型差异蛋白质并进行GO、KEGG、亚细胞定位与功能注释。瘤内注射1×106 TCID50 MV-Hu191干预4T1细胞移植瘤模型小鼠，流式细胞术检测小鼠脾组织中T细胞亚群，ELISA法检测小鼠血清TNF-α和IL-6含量。结果：体外实验结果表明，MV-Hu191具有抑制MDA-MB-231和4T1细胞增殖的作用，差异均具有统计学意义（P<0.01）。蛋白质组学分析结果显示，MV-Hu191作用MDA-MB-231细胞后明显上调蛋白质有38个、下调有12个；差异表达的蛋白质主要参与细胞黏附、信号受体激活、细胞代谢、应激反应等生物学过程，22个差异蛋白质亚细胞定位位于细胞外，KEGG功能分类显示与免疫调节功能相关的差异蛋白质最多且均为上调蛋白，包括C4A、C8B、SERPINF2、A2M、SERPINC1、CTSB、SERPING1、C5；PPI预测发现免疫相关差异蛋白与CD4、CD8、TNF-α及IL-6相互关联。体内实验结果显示，MV-Hu191干预组小鼠脾组织中CD4+ T细胞数量略高于对照组，但差异无统计学意义（P>0.05），CD4+/CD8+ T细胞比值明显高于对照组（P<0.05），血清TNF-α和IL-6含量显著上升（均P<0.01）。结论：MV-Hu191显著抑制MDA-MB-231、4T1细胞增殖及拮抗4T1细胞荷瘤小鼠成瘤性，其机制可能是MV-Hu191通过激活免疫效应分子实现抗肿瘤作用。

circNEIL3 regulates the proliferation, migration and invasion of breast cancer MDA-MB-231 cells through targeting miR-4784 / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：研究环状RNA nei样DNA糖化酶3（circNEIL3）和微小RNA（miR）-4784对乳腺癌细胞MDA-MB-231的增殖、迁移和侵袭的影响。方法：收集2018年1月至2019年12月在济南市中西医结合医院经组织病理诊断为乳腺癌并行手术切除的45例乳腺癌患者的癌组织和配对癌旁组织，qPCR法检测乳腺癌组织、癌旁组织中circNEIL3和miR-4784的相对水平。将circNEIL3的小干扰RNA（si-circNEIL3）、miR-4784模拟物、si-circNEIL3+miR-4784抑制物分别转染MDA-MB-231细胞，采用CCK-8法、平板克隆实验、划痕愈合实验、Transwell实验检测circNEIL3和miR-4784表达对细胞活力、克隆形成、迁移和侵袭的影响。双荧光素酶报告基因实验、RNA免疫沉淀（RIP）和RNA pull-down实验检测circNEIL3和miR-4784之间相互作用。结果：乳腺癌组织中circNEIL3呈高表达（P<0.05），miR-4784呈低表达（P<0.05）。干扰circNEIL3显著降低MDA-MB-231细胞活力、克隆形成数、划痕愈合率以及侵袭数（均P<0.05）。过表达miR-4784显著降低MDA-MB-231细胞活力、克隆形成数、划痕愈合率以及侵袭数（均P<0.05）。双荧光素酶报告基因实验、RIP和RNA pull-down实验均证实circNEIL3与miR-4784可直接结合，干扰circNEIL3能明显上调miR-4784表达（P<0.05），过表达circNEIL3能明显下调miR-4784表达（P<0.05）。抑制miR-4784表达部分逆转干扰circNEIL3对MDA-MB-231细胞活力、克隆形成、迁移和侵袭的抑制作用（P<0.05）。结论：干扰circNEIL3通过靶向上调miR-4784表达抑制MDA-MB-231细胞的增殖、迁移和侵袭。

The inhibitory effect of Licochalcone B on the triple negative breast cancer cell MDA-MB-231 and its mechanism / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨甘草查尔酮B（LCB）对三阴性乳腺癌（TNBC）MDA-MB-231细胞的抑制作用及其机制。方法: 常规培养MDA-MB-231细胞，用不同浓度LCB处理后，采用CCK-8法、免疫荧光法、FCM和WB法分别检测MDA-MB-231细胞的增殖活力、细胞核内DNA双链断裂标志物γ-H2AX的表达，以及细胞周期和周期调控、丝裂原活化蛋白激酶（MAPK）、内质网应激信号途径相关蛋白的表达水平。结果: LCB能显著抑制乳腺癌MDA-MB-231细胞的增殖活力（P<0.05），使γ-H2AX阳性细胞数和蛋白表达水平均显著升高（均P<0.05）、G2/M和S期的细胞数量均明显增加（均P<0.05）、MAPK家族主要成员细胞外调节激酶1/2（ERK1/2）和p38MAPK蛋白的磷酸化水平均显著上调（均P<0.05），还使内质网应激途径相关蛋白Bip、ATF4和CHOP的表达均显著上调（均P<0.05）。结论: LCB能够显著抑制MDA-MB-231细胞的增殖活力、诱导DNA损伤和细胞周期阻滞于G2/M和S期，LCB对MDA-MB-231细胞的抑制作用可能与其激活MAPK和内质网应激信号通路相关。

Norcantharidin induces apoptosis through autophagosome accumulation in breast cancer MDA-MB-231 cells / 中国药科大学学报

@#The aim of this study was to investigate the effect of norcantharidin (NCTD) on the proliferation and apoptosis of triple-negative breast cancer cell line MDA-MB-231.Western blot was used to detect the effect of NCTD on the expression levels of apoptosis-related proteins Bax/Bcl-2, cleaved-PARP/PARP/PARP, cleved-caspase-9, cleaved-caspase-3 and MCL-1 in MDA-MB-231 cells.Also, the expression levels of autophagy-related proteins LC3-II/LC3-I, Parkin and PINK1 in MDA-MB-231 cells were measured by Western blot.Flow cytometry was used to measure the effect of NCTD on the changes of mitochondrial membrane potential and mitochondrial reactive oxygen species (ROS).The effect of NCTD on autophagy flow in cells expressing mCherry-EGFP-LC3 was detected by a confocal microscope.Moreover, the effects of NCTD combined with chloroquine (CQ) or 3-methyladenine (3-MA) on the apoptosis of MDA-MB-231 cells were detected by flow cytometry.The results showed that NCTD significantly increased the expression levels of Bax/Bcl-2, cleaved-PARP/PARP, cleaved-caspase-9, cleasved-caspase-3 and LC3-II/LC3-I proteins, and promoted the mitochondrial translocation of Parkin, and blocked the autophagic flow in MDA-MB-231 cells. Moreover, NCTD combined with CQ accelerated apoptosis, while NCTD combined with 3-MA decreased apoptosis.These results suggest that NCTD can induce autophagy accumulation and lead to apoptosis of MDA-MB-231 cells.

Morphine counteracts the effects of paclitaxel in triple-negative breast cancer cells

Background & objectives: Several studies have provided evidence that opioids may play a role in cancer recurrence and metastasis. Multiple research data indicate that morphine can act as a proliferative or suppressive agent on tumour cells depending on the applied concentration. Therefore, this study was aimed to investigate whether the presence of clinically relevant concentrations of morphine has any effect on the efficacy of paclitaxel, a widely used chemotherapeutic drug, on the viability and apoptosis of human triple-negative breast cancer cell line. Methods: MDA.MB.231 cells were treated with paclitaxel in the presence or absence of morphine and examined for cell proliferation by the MTT assay. In addition, the effect of morphine on paclitaxel- induced apoptosis was investigated by flow cytometric assay and by the ratio of Bax/Bcl-2 mRNA expression levels with quantitative real-time (qRT)-PCR. Results: Morphine significantly increased the proliferation of breast cancer cells at low concentrations (0.1-2.5 ?M) but higher concentrations showed cytotoxic effect. Pre-treatment with 0.1 or 1 ?M of morphine decreased the paclitaxel-induced cytotoxicity, the proportion of apoptotic cell, and the ratio of Bax/Bcl-2 mRNA expressions. Interpretation & conclusions: Our data suggest that morphine promotes breast cancer cell viability at clinically relevant plasma concentrations and reduces the apoptotic effect of paclitaxel. This interaction may be very important in clinical settings; however, more studies are needed to explore the plausible mechanisms of interaction and to correlate such findings through in vivo animal studies as well as clinically.

Piperine suppresses growth and migration of human breast cancer cells through attenuation of Rac1 expression

Objective: To investigate the effect of piperine on human breast cancer cells. Methods: The effect of piperine on proliferation and migration of human breast cancer cells, MCF-7 and MDA-MB-231, was investigated using colony formation assays, wound healing assays, Matrigel migration assays, flow cytometry, RT-qPCR, and Western blotting assays. Results: Piperine inhibited the growth of MCF-7 and MDA-MB-231 cells and suppressed colony formation. Cell reduction at the G 0 / G 1 phase and cell arrest at the G 2 /M phase were observed in breast cancer cells. However, the significant effect was only demonstrated in MDA-MB-231 cells. Moreover, cancer cell migration was suppressed by piperine at low concentration. RT-qPCR and Western blotting assays showed that piperine downregulated Rac1 gene and protein expression. Conclusions: Piperine could inhibit growth and migration of breast cancer cells by reducing Rac1 gene and protein expression.

Cytotoxic evaluation and LC-MS/MS analysis of aerial parts of Eryngium kotschyi Boiss. grown in Turkey

Abstract Increasing biological activity and phytochemical investigations on Eryngium species showed its potential as pharmaceutical approach. Eryngium kotschyi Boiss. is one of the species of Eryngium genus and is endemic to Turkey. It is known that this plant is traditionally used in the South-western part of Turkey for the treatment of various diseases. This study focuses on cytotoxic activities of methanol extract and ethyl acetate, n-butanol and water sub-extracts from E. kotschyi in A549, COLO 205 and MDA-MB-231 cell lines by Sulforhodamin B assay and qualitative and quantitative determination of phytochemical constituents in active extract by LC-MS/MS. From the result of the study, it was seen that E. kotschyi ethyl acetate (EKE) sub-extract showed the strongest cytotoxic effect with the low IC50 values (50.00; 31.96 and 22.26 µg/mL in A549; COLO 205 and MDA-MB-231 cells at 48 h, respectively). Preliminary examination of the mass spectrums revealed the presence of 15 phytochemical compounds in active sub-extract and 7 of them was quantified. According to quantitative analyses the main compounds of EKE sub-extract were rosmarinic acid (485.603 µg/mgextract), chlorogenic acid (62.355 µg/mgextract) and caffeic acid (59.266 µg/mgextract). Moreover, this preliminary study on inhibitory activity of EKE sub-extract suggests further toxicologic investigations and detailed investigation on cytotoxic effect of various combinations of determined compounds

The Potentiality of Citral in Targeting Breast Cancer Multicellular Tumour Spheroids (MTS)

@#Introduction: As the high incidence of breast cancer has a profound impact on a global scale, there is a critical need to improve the clinical outcome of the patients, including efforts to utilize bioactive natural products as treatment or preventive measures. Citral, the essential oil of lemongrass has been reported to possess cytotoxicity in breast cancer cell line . The aim of present study was to determine the capability of citral in targeting aldehyde dehydrogenase-positive (ALDH+) cells in breast cancer cells. Methods: Both MCF-7 and MDA-MB-231 cells were cultured in serum-free media to generate multicellular tumour spheroids for the evaluation of citral as an antiproliferative agent. The cells were treated with identified IC50 (50±4.30 µM and 56±3.17 µM of citral, respectively) to investigate the cytotoxicity of citral. Staining using Propidium Iodide (PI) and Hoechst 33342 was carried out to determine cell proliferation and viability. Finally, ALDH+ cells were quantified via ALDEFLUOR assay. Analysis of differences was carried out by analysis of variance (ANOVA) and independent t-test with p<0.05 considered statistically significant. Results: The size of spheroids in both cancer cell lines were reduced after treatment with the citral. PI and Hoechst 33342 staining also revealed that citral gave rise to a mixture of cells that are normal and undergoing apoptosis and necrosis. ALDEFLUOR assay analysis revealed citral significantly (p <0.05 ) inhibited the population of ALDH+ cells in MCF7 cells. Conclusion: It was demonstrated that citral reduced the ALDH+ cell population in MCF7 breast cancer spheroids by inhibiting the ALDH activity.

miR-32-5p regulates the biological behaviors of breast cancer MDA-MB-231 cells by targeting the expression of Dickkopf-related protein 3 / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：通过生物信息学手段筛选乳腺癌中差异表达的关键miRNA及其靶基因，干预其在乳腺癌细胞中的表达并观察对乳腺癌细胞功能的影响。方法：利用GEO数据库筛选在乳腺癌中差异表达的miRNA，ENCORI数据库验证差异miRNA的表达，以选定最显著的差异表达miRNA为研究对象；利用Starbase、miRDB和miRWalk数据库预测miR-32-5p的靶基因，利用DAVID数据库对靶基因进行GO分析和KEGG分析，利用String数据库联合Cytoscape3.6.2软件进行PPI网络分析及核心基因的筛选，从核心基因中选择相互联系紧密“度值”最显著的Dickkopf相关蛋白3（DDK3）基因进行后续实验。qPCR检测miR-32-5p在人正常乳腺细胞 MCF10A和人乳腺癌细胞MCF7、MDA-MB-231、MDA-MB-453细胞中的表达。向MDA-MB-231细胞中转染miR-32-5p mimic、miR-32-5p inhibitor及各自的对照（NC）序列，分别用CCK-8法、流式细胞术和Transwell实验检测过表达或抑制miR-32-5p对细胞增殖、凋亡和侵袭的影响。结果：从GEO数据库中获取的两个数据集共识别出两个差异miRNA，ENCORI数据库验证差异miRNA的表达发现miR-32-5p的表达水平与GEO数据库的结果一致，故选择其进行研究；预测得到198个miR-32-5p潜在的靶基因并鉴定出10个核心基因（DKK3、WNT2B、SFRP5、SFRP2、SFRP1、LRP6、WNT6、KREMEN1、NEDD4L、TRIP12），其中DKK3的度值最大可能在乳腺癌中较为重要，于是选择miR-32-5p/DKK3轴进行后续研究。miR-32-5p在3种乳腺癌细胞中的表达水平显著高于正常乳腺细胞（均P<0.01），其中以MDA-MB-231细胞中表达最高。双荧光素酶基因报告实验验证了miR-32-5p与DKK3基因的靶向结合及其对后者表达的负向调控。转染miR-32-5p mimic、miR-32-5p inhibitor后成功提高或抑制了MDA-MB-231细胞中miR-32-5p的表达。与对照组相比，过表达miR-32-5p可抑制MDA-MB-231细胞的凋亡而促进细胞增殖和侵袭（P<0.05或P<0.01），敲低miR-32-5p则起相反的作用（均P<0.01）。结论：miR-32-5p/DKK3轴可能是影响乳腺癌发生发展的关键通路，过表达miR-32-5p能够抑制乳腺癌MDA-MB-231细胞的凋亡而促进细胞增殖和侵袭。

Effect of Curdione on MDA-MB-231 Cell Cycle and Apoptosis / 中国实验方剂学杂志

Objective:To investigate the effects of curdione on the proliferation， apoptosis and cell cycle of triple negative breast cancer cell line MDA-MB-231. Method:MDA-MB-231 cells were cultured<italic> in vitro</italic> with capecitabine （positive control） and curdione at different concentrations （125， 250， 500， 1 000， and 2 000 μmol·L^-1）， respectively， for detecting their viability using the cell counting kit-8 （CCK-8） at 24 and 48 h. Three effective inhibitory concentrations （250， 500， and 1 000 μmol·L^-1） against cell proliferation were selected for subsequent experiments. The effect of curdione on cell cycle was determined by flow cytometry combined with propidium iodide （PI） staining. After the set-up of high-concentration （2 000 μmol·L^-1） group， the effect of curdione on cell mitochondrial membrane potential was measured by JC-1（5，5，6，6-tetrachloro-1，1，3，3-tetraethylbenzimidazolylcarbocyanine iodide） staining， followed by the detection of cell apoptosis by flow cytometry combined with Annexin V-FITC/PI double staining. The changes in cell cycle status and apoptosis-related protein expression following curdione intervention were assayed by Western blot. Result:Compared with the blank control， curdione at 250， 500， 1 000， and 2 000 μmol·L^-1significantly inhibited the proliferation of MDA-MB-231 cells （<italic>P</italic><0.01）， exhibiting a concentration- and time-response relationship. The half maximal inhibitory concentration （IC₅₀） values at 24 and 48 h were 1 607 and 1 401 μmol·L^-1， respectively. Curdione at 250， 500， and 1 000 μmol·L^-1 arrested cells in G₁ phase. Curdione at 250 μmol·L^-1had no effect on cell mitochondrial membrane potential， which， however， declined significantly in the 500， 1 000， and 2 000 μmol·L^-1groups （<italic>P</italic><0.05， <italic>P</italic><0.01）. Curdione at 250， 500， and 1 000 μmol·L^-1obviously increased the proportion of apoptotic cells （<italic>P</italic><0.05， <italic>P</italic><0.01）. Curdione at each concentration elevated the Bcl-2-associated X protein （Bax）/B-cell lymphoma 2 （Bcl-2） ratio （<italic>P</italic><0.05， <italic>P</italic><0.01）， but did not change the cysteinyl aspartate-specific protease-3 （Caspase-3） expression. The protein expression levels of Caspase-9， cleaved Caspase-9， cleaved Caspase-3， p53， and p21 were up-regulated （<italic>P</italic><0.05）. Conclusion:A certain concentration of curdione inhibits the proliferation of MDA-MB-231 cells， which may be related to its efficacy in arresting cell cycle and inducing apoptosis.

Effect of Shugan Jianpi Jiedu Prescription Medicated Serum on Proliferation， Migration and Invasion of Triple-negative Breast Cancer MDA-MB-231 Cells Based on PI3K/Akt/mTOR Signaling Pathway / 中国实验方剂学杂志

Objective:To study the efficacy and mechanism of Shugan Jianpi Jiedu prescription （SJJ） in the treatment of triple-negative breast cancer through <italic>in vitro</italic> cell experiments. Method:The following groups were set up in this study： a normal serum group，a pirarubicin group，and low-，medium-， and high-dose SJJ-medicated serum groups. Twenty SD rats were randomly divided into four groups and administered with SJJ solution （16.8，8.2，4.05 g·kg^-1） and normal saline （equal volume） according to the body surface area to prepare serum. MDA-MB-231 cells were treated separately. The proliferation， migration and invasion of MDA-MB-231 cells were detected by the cell counting kit-8（CCK-8），wound healing assay and transwell cell invasion assay. The phosphoinositide 3-kinase （PI3K），protein kinase B （Akt）， and mechanistic target of rapamycin （mTOR） protein expression levels in MDA-MB-231 cells were tested by the Western blot. Result:The cell proliferation in the three different doses of medicated serum groups and the pirarubicin positive control group was significantly inhibited as compared with that in the normal serum group（<italic>P</italic><0.01），and there was no statistical difference for this between the medium/high dose medicated serum group and the pirarubicin positive control group.The wound healing in the SJJ-medicated serum groups and the pirarubicin group was slowed down as compared with that in the normal serum group （<italic>P</italic><0.01），and the effect in the SJJ-medicated serum groups was weaker than that in the pirarubicin group （<italic>P</italic><0.05，<italic>P</italic><0.01）. The number of cells invading the lower transwell chamber was decreased as compared with that in the normal serum group （<italic>P</italic><0.01），and there was no statistical difference between the medium-/high-dose SJJ-medicated serum groups and the pirarubicin group. Western blot results showed that 48 h after treatment，the PI3K，Akt， and mTOR expression levels in the cells of SJJ-medicated serum groups and the pirarubicin group were lower than those of the normal serum group（<italic>P</italic><0.01）. Conclusion:The SJJ-medicated serum could inhibit the proliferation， migration and invasion of MDA-MB-231 cells presumedly by down-regulating the protein expression levels in the PI3K/Akt/mTOR signaling pathway.

Inhibitory Effect and Mechanism of Jingulian Capsule on Human Breast Cancer MDA-MB-231 Cells / 中国实验方剂学杂志

Objective:To observe effect of Jingulian capsule on the proliferation of human breast cancer MDA-MB-231 cells and investigate its action mechanism against triple negative breast cancer （TNBC）. Method:The ingredients of Jingulian capsule were identified by ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）. The inhibitory effect of Jingulian capsule at different doses （0.125，0.25，0.5，1，and 2 g·L^-1） against the proliferation of MDA-MB-231 cells were detected by methyl thiazolyl tetrazolium （MTT） assay. After treatment for 24 h， the morphological changes in nuclear apoptosis of MDA-MB-231 cells were detected by Hoechst 33258 staining. The effect of different concentrations of Jingulian capsule on the apoptosis and cycle of MDA-MB-231 cells after different treatment time were determined by flow cytometry. The protein expression levels of Poly-ADP-ribose polymeras （PARP）， proto-oncogene c-Myc， cyclin B₁， and phosphorylated extracellular signal-regulated kinase （p-ERK） in each group were assayed by Western blot. Result:A total of 113 compounds in Jingulian capsule were identified by UPLC-MS/MS. As revealed by MTT assay，compared with blank group，Jingulian capsule （0.125，0.25，0.5，1，2 g·L^-1） significantly inhibited viability of MDA-MB-231 cells （<italic>P</italic><0.01）， with the half maximal inhibitory concentration （ IC₅₀） of（0.13±0.02）g·L^-1. According to flow cytometry，compared with the blank group，Jingulian capsule at 1 g·L^-1 significantly induced the apoptosis of MDA-MB-231 cells （<italic>P</italic><0.05）and Jingulian capsule at 0.5， 1 g·L^-1 obviously increased the number of MDA-MB-231 cells in S phase （<italic>P</italic><0.05，<italic>P</italic><0.01）. The results of Western blotting demonstrated that the protein expression levels of PARP，c-Myc，and cyclin B₁ in 0.5， 1 g·L^-1Jingulian capsule groups were remarkably down-regulated as compared with those in the blank group（<italic>P</italic><0.01）， and the protein expression level of p-ERK in 1 g·L^-1Jingulian capsule group was also down-regulated （<italic>P</italic><0.01）. Conclusion:Jingulian capsule is able to inhibit the proliferation of MDA-MB-231 cells，induce S phase cell cycle arrest， and promote their apoptosis， which may be related to the inactivation of the MAPK signaling pathway.

TM9SF2 promotes proliferation and metastasis of triple negative breast cancer cell line MDA-MB-231 / 中国药科大学学报

@#The aim of this study was to investigate the effect of transmembrane 9 superfamily protein member 2 (TM9SF2) in proliferation and migration of triple negative breast cancer cell line MDA-MB-231.The expression of TM9SF2 in triple negative breast cancer cell line MDA-MB-231 and nontumorigenic mammary epithelial cell line MCF-10A were measured by Western blot. MDA-MB-231 cells were treated with siRNA-TM9SF2 to knock-down the expression of TM9SF2. The effect of silencing TM9SF2 was measured with Western blot.The proliferation of cells was tested by MTS,and the migration was measured with Transwell and wound-healing assay.Proteins related to proliferation (PI3K,AKT,SRC and ERK) and migration (Snail,Slug and N-cadherin) were measured with Western blot.Protein expressions of TM9SF2 was better improved in triple negative breast cancer MDA-MB-231 cell line than MCF-10A.Compared with the control group, the siRNA-TM9SF2 infected group had lower expressions of PI3K, Snail, Slug and N-cadherin, and at the same time phosphorylation of AKT was decreased. The results suggest TM9SF2 can promote the proliferation and metastasis of triple negative breast cancer MDA-MB-231 cell line.

Effects of Urtica dioica extract on malignant biological behaviors of breast cancer cells and its possible mechanism / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨大荨麻提取物对乳腺癌细胞增殖、凋亡和细胞周期的影响，并初步探讨其可能的作用机制。方法：用不同质量浓度的大荨麻提取物（0、1、2、4、8、16、32、64 mg/ml）处理乳腺癌细胞MCF-7和MDA-MB-231 24 h，MTT法检测细胞增殖活力，选择中位抑制浓度附近的浓度（5和10 mg/ml）作为给药浓度分别处理MCF-7和MDA-MB-231细胞24 h后，平板克隆形成实验和流式细胞术分别检测大荨麻提取物对乳腺癌细胞增殖、周期和凋亡的影响，WB法检测对细胞周期和凋亡相关蛋白以及PI3K/AKT信号通路相关蛋白表达的影响。在MCF-7细胞用5 mg/ml大荨麻提取物处理的同时转染过表达AKT质粒（大荨麻+AKT组），转染空载质粒为对照组（大荨麻+vec组），WB法检测过表达效率，比较过表达AKT对细胞增殖、周期和凋亡的影响。结果：各大荨麻提取物处理组MCF-7和MDA-MB-231细胞增殖活力均显著低于对照组（P<0.05或P<0.01）；与对照组比较，5或10 mg/ml大荨麻处理组乳腺癌细胞的克隆形成数显著减少，G0/G1期细胞占比和凋亡率显著增加（P<0.05或P<0.01），P21、BAX蛋白表达显著升高而Cyclin D1、CDK4、Bcl2蛋白以及p-PI3K、p-AKT蛋白表达显著降低（P<0.05或P<0.01）。大荨麻+AKT组p-AKT和AKT蛋白表达显著高于大荨麻+vec组，克隆数、S期和G2/M期细胞占比均高于大荨麻+vec组（P<0.05或P<0.01），G0/G1期细胞占比和凋亡率低于大荨麻+vec组（P<0.05或P<0.01）。结论：大荨麻提取物可以抑制乳腺癌细胞增殖、促进凋亡且阻滞细胞在G0/G1期，其作用机制可能与抑制PI3K/AKT信号通路相关。

Purple sweet potato anthocyanin regulates the proliferation, migration and invasion of breast cancer MDA-MB-231 cells through circ_0003998/miR-145 axis / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨紫甘薯花色苷（purple sweet potato anthocyanin, PSPA）是否通过circ_0003998/miR-145轴调控乳腺癌MDA-MB-231细胞的增殖、迁移和侵袭。方法：选用乳腺癌MDA-MB-231细胞，将其分为对照组，200、400和800 μg/ml PSPA组，pcDNA组、pcDNA-circ_0003998组、si-NC组、si-circ_0003998组、si-circ_0003998+anti-miR-145组、PSPA+pcDNA组、PSPA+pcDNA-circ_0003998组和PSPA+anti-miR-145组。用qPCR法检测细胞中circ_0003998和miR-145的表达，CCK-8法、Transwell小室法分别检测转染前后细胞的增殖、迁移和侵袭能力，WB法检测细胞中Ki-67、MMP-2和MMP-9蛋白的表达。用双荧光素酶报告基因实验验证circ_0003998与miR-145的靶向关系。结果：与对照组比较，各剂量PSPA组MDA-MB-231细胞的增殖抑制率、miR-145表达水平均显著升高（均P<0.01），Ki-67、MMP-2、MMP-9蛋白和circ_0003998的表达水平、细胞迁移和侵袭细胞数均显著降低（均P<0.01），并呈现浓度依赖性。circ_0003998可以靶向负调控miR-145的表达。敲减circ_0003998后，MDA-MB-231细胞的增殖抑制率、miR-145表达水平显著升高，Ki-67、MMP-2和MMP-9蛋白表达水平、细胞迁移和侵袭细胞数均显著减少（均P<0.01）。共转染si-circ_0003998和anti-miR-145则可逆转敲减circ_0003998表达对MDA-MB-231细胞增殖、迁移和侵袭的抑制作用，过表达circ_0003998或抑制miR-145表达可逆转PSPA对MDA-MB-231细胞增殖、迁移和侵袭的抑制作用。结论：PSPA通过circ_0003998/miR-145轴抑制乳腺癌MDA-MB-231细胞的增殖、迁移和侵袭。

Effect and Its Mechanism of Plantago asiatica Polysaccharide on the Proliferation ，Migration and Invasion of Breast Cancer Cells / 中国药房

OBJECTIVE：To study the effects of Plantago asiatica polysaccharide on the proliferation ，migration and invasion of breast cancer cells ，and to investigate its mechanism preliminarily. METHODS ：Using human breast cancer cell MDA-MB- 231 as subjects ，MTT method was adopted to detect the effects of different concentrations of P. asiatica polysaccharide（8，16，32，64 mg/L）on the cell proliferation ability ，and survival rate of the cells was calculated. Scratch test and Transwell invasion test were used to detect the effects of different concentrations of P. asiatica polysaccharide（8，16 mg/L）on cell migration ability and invasion ability. Western blot assay was used to detect the expression of epithelial-mesenchymal transition （EMT）-related proteins [matrix metalloproteinase- 2（MMP-2），MMP-9，E-cadherin，N-cadherin，vimentin]. RESULTS ：Results of MTT assay showed that survival rate of the cells in 32，64 mg/L P. asiatica polysaccharide groups were significantly lower than control group （P＜0.05 or P＜0.01），so that 8，16 mg/L，which did not affect the cell survival rate ，were used as the follow-up drug concentrations. Compared with control group ，relative mobility （12，24 h），relative invasion rate and relative expression of MMP- 2，MMP-9， N-cadherin and vimentin protein were decreased significantly in 8，16 mg/L P. asiatica polysaccharide groups （P＜0.05 or P＜ 0.01），while relative expression of E-cadherin protein was increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：P. asiatica polysaccharide can inhibit the proliferation of breast cancer cells MDA-MB- 231，and inhibit the migration and invasion of the cells by regulating the expression of metastasis and EMT-related proteins.

Escape from abluminal LRP1-mediated clearance for boosted nanoparticle brain delivery and brain metastasis treatment

Breast cancer brain metastases (BCBMs) are one of the most difficult malignancies to treat due to the intracranial location and multifocal growth. Chemotherapy and molecular targeted therapy are extremely ineffective for BCBMs due to the inept brain accumulation because of the formidable blood‒brain barrier (BBB). Accumulation studies prove that low density lipoprotein receptor-related protein 1 (LRP1) is promising target for BBB transcytosis. However, as the primary clearance receptor for amyloid beta and tissue plasminogen activator, LRP1 at abluminal side of BBB can clear LRP1-targeting therapeutics. Matrix metalloproteinase-1 (MMP1) is highly enriched in metastatic niche to promote growth of BCBMs. Herein, it is reported that nanoparticles (NPs-K-s-A) tethered with MMP1-sensitive fusion peptide containing HER2-targeting K and LRP1-targeting angiopep-2 (A), can surmount the BBB and escape LRP1-mediated clearance in metastatic niche. NPs-K-s-A revealed infinitely superior brain accumulation to angiopep-2-decorated NPs-A in BCBMs bearing mice, while comparable brain accumulation in normal mice. The delivered doxorubicin and lapatinib synergistically inhibit BCBMs growth and prolongs survival of mice bearing BCBMs. Due to the efficient BBB penetration, special and remarkable clearance escape, and facilitated therapeutic outcome, the fusion peptide-based drug delivery strategy may serve as a potential approach for clinical management of BCBMs.

Cytotoxicity of oleanane type triterpene from leaf extract of Pterospermum acerifolium (in vitro) and theoretical investigation of inhibitory signaling pathway / 中草药·英文版

Objective: To evaluate the cytotoxic activity of taraxerol isolated from the leaves of Pterospermum acerifolium and its EtOH extract against human breast, colon, and lung cancer cell lines and docking studies. Methods: The structures of the isolated compounds were elucidated by several spectroscopic methods such as

Inhibitory effect of disulfiram combined with cisplatin on triple negative breast cancer MDA-MB-231 cells and its mechanism / 吉林大学学报(医学版)

Objective: To observe the inhibitory effect of combination of disulfiram (DSF) and cisplatin (CDDP) on the proliferation of triple-negative breast cancer (TNBC) MDA-MB-231 cells, and to explore its possible mechanism. Methods: The MDA-MB-231 cells in the logarithmic phase were divided into blank control group. DSF group. CDDP group and combination groups (60/xmol • L 1 CDDP+1. 2 and 4/imol • L ' DSF). MTT method was used to detect the survival rates of the MDA-MB-231 cells in various groups and flow cytometry was used to detect the apoptotic rates of the MDA-MB-231 cells in various groups, flow cytometry was used to detect the level of active oxygen ( ROS ) in the MDA-MB-231 cells in various groups, and Inductively Coupled Plasma-MassSpectrometry (ICP-MS) was used to detect the levels of Pt in the MDA-MB-231 cells in various groups. Results: The MTT method and flow cytometry results showed that after treated for 72 h. compared with 60/xmol • L ' CDDPgroup, the survival rates of the MDA-MB-231 cells in combination 1 group (60/xmol • L CDDP+1/xmol • L 1 DSF), combination 2 group (60/xmol • L 'CDDP+ 2/xmol • L 1 DSF) and combination 3 group (60/xmol • L 'CDDP+4/xmol • L 'DSF) were decreased ( P<0. 05). After treated for 24 h. compared with 60/x mol • L ' CDDP group and 2/x mol • L DSF group, the apoptotic rate of the MDA-MB-231 cells in combination group (60/xmol • L 'CDDP+2/xmol • L ' DSF) was increased ( P<0. 05). Compared with CDDP group, the level of ROS in the MDA-MB-231 cells in combination group (60/xmol • L 'CDDP+2/xmol • L 1 DSF) was increased (P<0. 05). The 1CP-MS results showed that compared with CDDP group, the level of Pt in the MDA-MB-231 cells in combination group (20/xmol • L 1CDDP+ 0.625/xmol • L ' DSF) was increased (P< 0.05). Conclusion: The combination of DSF and CDDP can significantly inhibit the proliferation of MDA-MB-231 cells, and its possible mechanism may be related to inhibiting the growth of tumor cells by increasing the ROS level and Pt content in the MDA-MB-231 cells.

Network analysis on Rab7a gene in breast cancer MDA-MB-231 cells / 吉林大学学报(医学版)

Objective: To investigate the changes of expressions of the related genes in the triple-negative breast cancer (TNBC) MDA-MB-231 cells after knockout of Rab7a gene, and to elucidate the roles of Rab7a-related genes in TNBC. Methods: The breast cancer ZR-75-30, MCF-7, T-47D, MDA-MB-231 and HCC-1937 cells in the logarithmic phase were selected. The expression levels of Rab7a protein in the breast cancer ZR-75-30, MCF-7, T-47D, MDA-MB-231, and HCC-1937 cells were detected by Western blotting method. The Rab7a gene in the MDA-MB-231 cells was knockout with lentivirus and the cells were divided into negative control group and Rab7a knockout group. According to the four different knockout Rab7a sequences, the Rab7a knockout group was divided into KD1 group, KD2 group, KD3 group and KD4 group. The knockout efficiencies of Rab7a gene in the MDA-MB-231 cells in various groups were detected by qPCR method, the differential genes in the highest knockout efficiency group (KD2 group) and negative control group were detected by full gene expression microarray, and the interaction between Rab7a gene and other genes was analyzed by integrated pathway analysis (IPA) software. Results: The Rab7a protein was expressed in the TNBC MDA-MB-231 cells. Compared with negative control group, the knockout efficiency of Rab7a gene in the MDA-MB-231 cells in KD2 group was the highest (P