BRD4 inhibitor and histone deacetylase inhibitor synergistically inhibit the proliferation of gallbladder cancer in vitro and in vivo

Gallbladder cancer (GBC) is the most common malignancy of the bile duct and has a high mortality rate. Here, we demonstrated that BRD4 inhibitor JQ1 and histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) synergistically inhibited the GBC cells in vitro and in vivo. Our results showed that cotreatment with JQ1 and SAHA significantly inhibited proliferation, cell viability and metastasis, and induced apoptosis and G2/M arrest in GBC cells, with only minor effects in benign cells. In vivo, tumor volumes and weights of GBC xenograft models were significantly decreased after treatment with JQ1 or SAHA; meanwhile, the cotreatment showed the strongest effect. Further study indicated that the above anticancer effects was associated with the downregulation of BRD4 and suppression of PI3K/AKT and MAPK/ERK pathways. These findings highlight JQ1 and SAHA as potential therapeutic agents and their combination as a promising therapeutic strategy for GBC.     

MEK inhibitor effective against proliferation in breast cancer cell

The targeted small-molecule drug AZD6244 is an allosteric, ATP-noncompetitive inhibitor of MEK1/2 that has shown activity against several malignant tumors. Here, we report that AZD6244 repressed cell growth and induced apoptosis and G1-phase arrest in the breast cancer cell lines MDA-MB-231 and HCC1937. Using microRNA (miRNA) arrays and quantitative RT-PCR, we found that miR-203 was up-regulated after AZD6244 treatment. In accordance with bioinformatics and luciferase activity analyses, CUL1 was found to be the direct target of miR-203. Furthermore, miR-203 inhibition and CUL1 overexpression reversed the cytotoxicity of AZD6244 on the MDA-MB-231 and HCC1937 cells. Collectively, our data indicate that miR-203 mediates the AZD6244-induced cytotoxicity of breast cancer cells and that the MEK/ERK/miR-203/CUL1 signaling pathway may participate in this process.     

miR-195 Targets HDGF to inhibit proliferation and invasion of NSCLC cells

Non-small cell lung cancer (NSCLC) is one of the most common causes of cancer-related death worldwide. MicroRNAs (miRNAs) play critical roles in the development and progression of NSCLC. miR-195 acts as a tumor suppressor in several cancers, however, its role in NSCLC is not well understood. Herein, we found that miR-195 was significantly decreased in both NSCLC tissues and cell lines. Forced expression of miR-195 significantly suppressed proliferation, migration, and invasion of NSCLC cells. Hepatoma-derived growth factor (HDGF) was identified as a target of miR-195 in NSCLC cells. Overexpression of HDGF dramatically abolished the tumor suppressive role of miR-195 in NSCLC cells. Our results demonstrated a tumor suppressive role of miR-195 in NSCLC, and suggested a potential therapeutic target for NSCLC.     

PI3K抑制剂ZSTK-474对结直肠癌细胞增殖和侵袭的影响

目的:探讨PI3K抑制剂ZSTK-474对结直肠癌细胞增殖和侵袭的影响。方法:采用不同浓度（0、1、2、4、6、8和10 μmol/L）ZSTK-474处理结直肠癌HT-29和HCT-116细胞24 h和48 h,CCK8法检测细胞增殖活性并计算出ZSTK-474在两株结直肠癌细胞中的半数抑制浓度（IC50）。根据IC50的值选用浓度为4 μmol/L的ZSTK-474作用HT-29和HCT-116细胞24 h,同时设置加入等量的0.1%DMSO为对照。采用平板克隆形成实验检测细胞增殖,流式细胞仪检测细胞凋亡,qRT-PCR检测细胞PI3K、Akt的mRNA水平,Western blot 检测细胞PI3K、Akt的蛋白水平。结果:ZSTK-474可明显抑制HT-29和HCT-116细胞的增殖,且呈时间和剂量依赖性。ZSTK-474可抑制HT-29和HCT-116细胞的侵袭能力,但不影响其凋亡。ZSTK-474处理HT-29和HCT-116细胞24 h后,PI3K和Akt的mRNA和蛋白表达水平均显著下降。结论:ZSTK-474能够抑制结直肠癌细胞增殖,并且能抑制其侵袭。     

MicroRNA-183抑制结直肠癌肿瘤细胞的增殖、侵袭及迁移

目的:探讨microRNA183 （miRNA-183）对结直肠癌肿瘤细胞增殖、侵袭及迁移的影响。方法:使用慢病毒技术,分别转染HT29细胞株,建立稳定过表达细胞株miRNA-183 mimics,抑制miRNA-183表达的稳定细胞株miRNA-183 inhibitor以及阴性对照组negative control。qRT-PCR检测各组细胞miRNA-183的表达,Western blot检测miRNA-183下游靶基因PDCD4蛋白水平表达变化,CCK8增殖实验检测各组细胞的增殖情况,同时以划痕实验和Transwell实验分析转染miRNA-183 mimics和miRNA-183 inhibitor对HT29细胞迁移、侵袭能力的影响。结果:与阴性对照组相比,miRNA-183 mimics组细胞增殖速度明显下降,其迁移和侵袭能力也显著降低,差异均具有统计学意义（P<0.05）;miRNA-183下游靶基因PDCD4蛋白水平表达明显增加。结论:miRNA-183可有效地抑制结直肠癌肿瘤细胞的增殖、侵袭及迁移。     

LSD1抑制剂抑制非小细胞肺癌A549细胞侵袭、迁移及上皮间质转化

目的:探究赖氨酸特异性组蛋白去甲基化酶1（lysine-specific demethylase 1,LSD1）抑制剂对非小细胞肺癌（non-small cell lung cancer,NSCLC）A549细胞侵袭、迁移及上皮间质转化的影响。方法:将体外培养的人非小细胞肺癌A549细胞分为对照组和LSD1抑制剂组,在细胞进入对数生长期时,向实验组细胞中分别加入不同浓度的LSD1抑制剂,培养24 h。CCK-8法用于细胞增殖检测,划痕实验用于细胞迁移检测,Transwell小室法用于细胞侵袭检测,裂解细胞提取总蛋白后通过蛋白质免疫印迹技术（Western blot,WB）检测A549细胞中上皮细胞标志物E型钙黏蛋白（E-cadherin）、间充质细胞标志物N型钙黏蛋白（N-cadherin）和波形蛋白（Vimentin）的表达水平。结果:与对照组相比,LSD1抑制剂组细胞增殖率、穿膜细胞数和划痕愈合率显著降低（P＜0.05）;LSD1抑制剂组相较于对照组,E-cadherin表达量升高,N-cadherin和Vimentin的表达量均降低（P＜0.05）。结论:LSD1抑制剂可抑制非小细胞肺癌A549细胞的侵袭、迁移及上皮间质转化。     

奥沙利铂联合替尼泊甙对胃癌BGC-823细胞的生长抑制和诱导凋亡的协同作用(英文)

Objective:The aim of this study was to investigate the synergistic effects of oxaliplatin and teniposide on proliferation and apoptosis of gastric cancer cell line BGC-823.Methods:MTT assay was carried to examine the inhibition rate of oxaliplatin and teniposide on gastric cancer cell line BGC-823 with various concentrations separately and associatively.The apoptosis rate of BGC-823 cells under the treatment of oxaliplatin or/and teniposide was examined by flow cytometry.The expression level of livin, an ap...     

蟾毒灵与紫杉醇协同抑制乳腺癌细胞增殖的机制

目的:研究蟾毒灵(Bufalin)与紫杉醇是否可协同抑制乳腺癌细胞增殖,探索可能的协同机制.方法:采用MTT法测定细胞活力、采用流式细胞术检测细胞凋亡,采用Western Blot法检测AKT、p-AKT、p38、p-p38、β-actin蛋白的表达.结果:MTT结果显紫杉醇和Bufalin可以剂量依赖方式抑制乳腺癌MCF-7细胞增殖.两药联合作用可协同抑制MCF-7细胞增殖.流式分析显40nmoL/L的紫杉醇作用24小时可诱导乳腺癌MCF-7发生明显的G2M期阻滞和12.5％的细胞凋亡,10nmol/L的Bufalin作用24小时未诱导明显的细胞凋亡,10nmol/L的Bufalin与40nmol/L的紫杉醇联合作用24小时后可诱导23.7％的MCF-7细胞凋亡.Western Blot分析显紫杉醇作用后导致AKT和p38的活化,Bufalin与紫杉醇联合作用后可抑制紫杉醇诱导的AKT活化,增强紫杉醇诱导的p38活化.结论:Bufalin可通过抑制AKT的活化、上调p38的活化与紫杉醇协同抑制乳腺癌细胞增殖,诱导细胞凋亡.     

紫花牡荆素抑制乳腺癌MCF-7细胞增殖与侵袭能力的研究

目的观察紫花牡荆素对乳腺癌MCF-7细胞株增殖与侵袭能力的影响并探讨其分子机制。方法应用四甲基偶氮唑蓝（MTT）比色法与侵袭实验检测紫花牡荆素对MCF-7细胞增殖与侵袭能力的影响,应用反转录PCR、Western blot法、Tunel法检测紫花牡荆素对基因表达、蛋白表达、细胞凋亡的影响。结果不同浓度紫花牡荆素均抑制MCF-7细胞增殖水平,同未加药对照组比较差异均有统计学意义（P〈0．05）,5、10、20μmol／L紫花牡荆素作用后,MCF-7细胞迁移数与未处理组相比分别降低20.3％、44．4％和50．3％（P〈0．05）。以10μmol／L紫花牡荆素处理后,凋亡细胞数增多,可以上调Bax与Caspase-3蛋白的表达水平,下调基质金属蛋白酶（MMP）-2与MMP-9的mRNA和蛋白表达水平。结论紫花牡荆素对于乳腺癌细胞恶性增殖与侵袭能力具有显著抑制作用。     

The HER-2-targeting antibodies trastuzumab and pertuzumab synergistically inhibit the survival of breast cancer cells

Trastuzumab (herceptin) and pertuzumab (Omnitarg, 2C4) are recombinant humanized monoclonal antibodies that target different extracellular regions of the HER-2 tyrosine kinase receptor. We explored combination effects of these agents in the HER-2-overexpressing BT474 breast cancer cell line. Trastuzumab and 2C4 synergistically inhibited the survival of BT474 cells, in part, because of increased apoptosis. Trastuzumab increased 2C4-mediated disruption of HER-2 dimerization with the epidermal growth factor receptor and HER-3. Combination drug treatment reduced levels of total and phosphorylated HER-2 protein and blocked receptor signaling through Akt but did not affect mitogen-activated protein kinase. These results suggest that combining HER-2-targeting agents may be a more effective therapeutic strategy in breast cancer rather than treating with a single HER-2 monoclonal antibody.     

A MEK inhibitor (U0126) markedly inhibits direct liver invasion of orthotopically inoculated human gallbladder cancer cells in nude mice

Primary cancer of the gallbladder is not unusual. Most cases of gallbladder cancer are found at an advanced stage, accompanied by the invasion to the liver, metastases to the lymph nodes and distant organs, and peritoneal dissemination. In this study, we first examined the effect of mitogen-activated protein kinase kinase (MEK) inhibitors on the production of matrix metalloproteinases (MMPs), urokinase-type plasminogen activator (uPA), and tissue inhibitors of metalloproteinases (TIMPs) in a human gallbladder cancer cell line, NOZ cells in vitro. MEK inhibitors (PD98059 and U0126) inhibited the production of MMP-2, MMP-9 and high MW uPA, and upregulated TIMPs (TIMP-1, TIMP-2 and TIMP-3). Subsequently, we examined the effect of U0126 on invasion and metastasis of orthotopically inoculated NOZ cells in nude mice. Direct liver invasion by cancer cells was detected in all of the mice in the control group, but in only one mouse in the U0126-treated group. Most of the primary tumors in the U0126-treated group expanded to the liver, but did not invade into the liver. Vessel invasion in the liver was evident in 4 out of 5 mice in the control group, but in only one mouse in the U0126-treated group. Lymph node metastases and peritoneal dissemination were recognized in all of the mice in both groups. All 5 mice in the U0126-treated group, and 4 out of 5 mice in the vehicle control group, had metastases in the lungs. The present results suggest that a MEK inhibitor, U0126, prolonged the survival of the mice with NOZ tumor by inhibiting direct liver invasion and vessel invasion of the cancer cells via down-regulation of the matrix degrading ability of the cancer cells.     

Akt抑制剂MK-2206在缺氧环境下对结肠癌SW480细胞增殖、侵袭的影响

目的:探讨Akt抑制剂MK-2206在缺氧环境下对人结肠癌SW480细胞增殖、侵袭的影响。方法:根据CCK-8实验结果选择CoCl2和MK-2206的浓度,最后分成空白组、MK-2206组、CoCl2组、MK-2206+CoCl2组;Transwell小室实验检测四组细胞的迁移和侵袭能力;RT-PCR法检测各细胞组中Akt、mTOR、HIF-1α的mRNA表达水平;Western blot技术检测各细胞组中Akt、p-Akt、mTOR、p-mTOR、HIF-1α蛋白表达的差异性情况。结果:CoCl2诱导的缺氧环境可以促进SW480细胞的侵袭、迁移(P＜0.05),同时能够促进HIF-1α的mRNA和蛋白表达(P＜0.05),但是低浓度范围内的CoCl2对SW480细胞增殖活性无明显影响(P＞0.05);MK-2206可以在体外的常氧和缺氧环境中抑制SW480细胞的增殖活性和侵袭、迁移能力(P＜0.05);MK-2206能在体外的常氧环境下抑制SW480细胞的Akt、mTOR、HIF-1α的mRNA表达(P＜0.05),缺氧环境无明显作用;同时MK-2206能够在常氧和缺氧环境中显著降低p-Akt、p-mTOR、HIF-1α的蛋白表达水平(P＜0.05)。 结论:Akt抑制剂MK-2206可以在体外的常氧和缺氧环境下抑制SW480细胞的增殖活性和侵袭能力,但缺氧环境可能会弱化MK-2206对SW480细胞迁移的抑制作用。     

MicroRNA-497 and bufalin act synergistically to inhibit colorectal cancer metastasis

The study aims to investigate the effect of microRNA-497 (miR-497) expression and bufalin treatment in regulating colorectal cancer invasion and metastasis. The expression of miR-497 in colorectal cancer cells with prior treatment with bufalin was determined using real-time quantitative PCR. The nude mouse abdominal aortic ring assay and the human umbilical vein endothelial cell (HUVEC) migration assays were used to measure the angiogenic effect of bufalin. The effect of both bufalin treatment and miR-497 overexpression on colorectal cancer metastasis was measured using an animal tumor model together with in vivo imaging. These results suggested: (1) In the HCT116 cells and HUVECs, proliferation was inhibited in a time-dependent and/or concentration-dependent manner following the administration of bufalin; (2) Bufalin inhibited cell migration in a concentration-dependent manner by cell motility assays; (3) In the aortic ring assay, administration bufalin to the aortic ring significantly promoted micro-angiogenesis of nude mouse abdominal aorta in a concentration-dependent and time-dependent manner; (4) miR-497 was upregulated in human colorectal cancer HCT116 cells treated with different concentrations of bufalin in a concentration-dependent manner; and (5) Combined application of bufalin and miR-497 significantly reduced metastatic lesions and reduced weight loss compared with bufalin alone and control groups in vivo. This study revealed that bufalin inhibited angiogenesis and regulated miR-497 expression and that bufalin and miR-497 acted in synergy to inhibit colorectal cancer metastasis, resulting in improved quality of life in a nude mouse model.     

PYCR1对胃癌细胞增殖和侵袭的影响

目的:探讨人吡咯啉-5-羧酸还原酶1(PYCR1)对胃癌细胞AGS增殖和侵袭的影响.方法:人胃癌细胞AGS转染敲减和过表达PYCR1的质粒,设置3种敲减序列,qRT-PCR和Western Blot检测PYCR1 mRNA和蛋白表达以验证敲减效率,并以敲减效率最好的进行后续实验,AGS细胞分为si-NC组、si-PYC...