南蛇藤提取物通过调控基质金属蛋白酶组及其抑制因子抑制人胃癌SGC-7901细胞侵袭转移的研究

目的探讨南蛇藤提取物(COE)对人胃癌SGC-7901细胞的侵袭和转移的影响及其分子机制。方法 MTT法检测COE对人胃癌SGC-7901细胞增殖的影响;Transwell小室实验检测COE对SGC-7901细胞侵袭和转移能力的影响。Western blotting检测经COE作用后的SGC-7901细胞中金属基质蛋白酶(MMPs)MMP2、MMP9和金属基质蛋白酶抑制因子(TIMPs)TIMP1、TIMP3的蛋白水平。RT-PCR检测经COE作用后的SGC-7901细胞中MMP2、MMP9、TIMP1、TIMP3的m RNA表达情况。结果 Transwell实验显示,经20、40、80 mg/L COE处理过的SGC-7901细胞穿膜数明显减少并呈一定的浓度依赖性。Western blotting结果显示,经20、40、80 mg/L COE处理过的SGC-7901细胞,MMP2、MMP9蛋白的表达水平明显降低,而TIMP1、TIMP3蛋白表达水平明显上调。RT-PCR结果显示,经20、40、80 mg/L COE处理过的SGC-7901细胞中MMP2和MMP9的m RNA水平有轻微升高趋势,而TIMP1和TIMP3的m RNA水平明显大幅上调(P0.001)。结论 COE能明显抑制人胃癌SGC-7901细胞的侵袭和迁移,其机制可能与下调MMP2、MMP9蛋白水平直接相关。而其作用的实现,很可能是通过直接上调TIMP1和TIMP3的m RNA转录水平从而提高TIMPs蛋白表达水平实现的。     

槐耳清膏抑制人前列腺癌PC3细胞增殖和侵袭作用及其机制研究

该研究探讨槐耳清膏对人前列腺癌PC3细胞增殖和侵袭的抑制作用及其相关分子机制。采用CCK-8法检测槐耳清膏对PC3细胞的增殖抑制作用。采用流式细胞术分析槐耳清膏给药后PC3细胞凋亡及细胞周期的变化。采用细胞划痕实验及Transwell侵袭实验分析给药后PC3细胞侵袭与迁移能力的变化。通过Western blot法,检测槐耳清膏给药后与侵袭迁移相关的EMT蛋白标志物表达水平的变化及MAPK信号通路中关键蛋白磷酸化水平的变化。结果显示,槐耳清膏可以明显抑制人前列腺癌PC3细胞的增殖。槐耳清膏8 g·L~(-1)作用于PC3细胞48 h后,细胞凋亡率较对照组明显升高,且细胞发生明显的S期阻滞。槐耳清膏给药后可以明显降低PC3细胞的侵袭与迁移能力,并且能够下调N-cadherin和TCF8/ZEB1的表达水平和上调E-cadherin的蛋白表达水平,表明槐耳清膏能够促进PC3细胞MET的发生,同时还发现MAPK信号通路中的关键蛋白JNK和ERK的磷酸化水平明显下降。因此,槐耳清膏能够抑制人前列腺癌PC3细胞增殖和迁移侵袭能力,这可能与其调控EMT及抑制MAPK信号通路有关。     

大蒜素对人子宫内膜癌Ishikawa细胞侵袭和迁移抑制作用的研究

目的:探索大蒜素对人子宫内膜癌Ishikawa细胞侵袭和迁移的抑制作用及其调控机制。方法:取对数生长期人子宫内膜癌Ishikawa细胞为受试细胞,设空白对照组(DMSO),大蒜素低、中、高剂量组(12.5、25、50μg/ml)和顺铂组(40μg/ml)。药物干预48 h后,CCK-8法检测细胞增殖抑制率,PI染色流式细胞术分析细胞周期分布,Transwell小室法、划痕实验检测细胞侵袭能力和迁移能力,Western blot法检测MMP-2、MMP-9、E-cadherin、PI3K、p-Akt蛋白表达。结果:经大蒜素中、高剂量或顺铂干预能够明显提高Ishikawa细胞增殖抑制率,提高处于G₀/G₁期Ishikawa细胞比例,降低Ishikawa细胞侵袭能力和迁移能力,下调MMP-2、MMP-9、E-cadherin、PI3K、p-Akt蛋白表达量,较空白对照组比较,差异均具有统计学意义(P<0.05)。结论:大蒜素对人子宫内膜癌Ishikawa细胞侵袭和迁移具有明显抑制作用,其机制可能与阻滞细胞周期进程而抑制细胞增殖,降低MMP-2...     

半枝莲抑制结肠癌细胞迁移和侵袭实验研究

目的:探讨半枝莲对结肠癌细胞迁移和侵袭的抑制作用及相关机制。方法:体外培养结肠癌SW620、CCL-228细胞系,用浓度为10 μmol/L半枝莲进行干预。将SW620、CCL-228细胞分为对照组(不做任何处理)、10 μmol/L半枝莲组、Rac1L61组(Rac1L61过表达质粒转染细胞)、联合组(Rac1L61过表达质粒转染细胞后加入10 μmol/L半枝莲)。实时细胞分析(RTCA)系统评估半枝莲对细胞的抑制作用,MTT法检测细胞活力,划痕实验检测细胞迁移,Transwell实验检测细胞侵袭,Western blotting法检测Rac1、p-PAK1、MMP2、MMP9蛋白表达水平。结果:10 μmol/L半枝莲组SW620、CCL-228细胞中Rac1、p-PAK1蛋白表达均低于对照组,差异有统计学意义(均P<0.05)。转染过表达Rac1质粒后,SW620、CCL-228细胞Rac1、p-PAK1蛋白表达均升高,而用10 μmol/L半枝莲干预可逆转这种效应。10 μmol/L半枝莲组SW620、CCL-228细胞侵袭和迁移能力均低于对照组,差异有统计学意义(均P<0.05)。转染过表达Rac1质粒后,SW620、CCL-228细胞侵袭和迁移明显增强,而用10 μmol/L半枝莲干预可逆转这种效应。10 μmol/L半枝莲组MMP2、MMP9蛋白表达低于对照组(均P<0.05)。转染过表达Rac1质粒后,SW620、CCL-228细胞MMP2、MMP9蛋白表达均升高,而用10 μmol/L半枝莲干预可逆转这种效应。结论:半枝莲通过调节Rac1/PAK1信号通路活性,减少MMP9、MMP2表达而降低结肠癌细胞迁移和侵袭能力。     

不同产地夏枯草中抗非小细胞肺癌成分科罗索酸的差异比较

目的:比较不同产地夏枯草中抗非小细胞肺癌活性成分科罗索酸的含量差异。方法:通过四甲基偶氮唑蓝(MTT)和吖啶橙(AO)/溴化乙啶(EB),评价科罗索酸对非小细胞肺癌细胞的增殖抑制以及凋亡作用;建立高效液相色谱法,比较14个产地夏枯草中科罗索酸的含量差异。结果:科罗索酸抑制SPC-A-1人非小细胞肺癌的生长增殖(IC50=1.05×10-6mol/L),并且诱导SPC-A-1细胞凋亡。14个产地夏枯草中科罗索酸的含量差异较大,山西产地的含量最高,河南-2产地的含量最低。结论:科罗索酸为夏枯草中的抗非小细胞肺癌的活性成分,不同产地夏枯草的科罗索酸含量差异较大。     

Cucurbitacin B inhibits the migration and invasion of breast cancer cells by altering the biomechanical properties of cells

Changes in cellular biomechanical properties affect cell migration and invasion. The natural compound Cucurbitacin B (CuB) has potent anticancer activity; however, the mechanism underlying its inhibitory effect on breast cancer metastasis needs further study. Here, we showed that low‐dose CuB inhibited adhesion and altered the viscoelasticity of breast cancer cells, thereby, reducing cell deformability. In vitro and in vivo experiments proved that CuB effectively inhibited the migration and invasion of breast cancer cells. Further studies have found that CuB downregulated the expression of F‐actin/vimentin/FAK/vinculin in breast cancer cells, altering the distribution and reorganization of cytoskeletal proteins in the cells. CuB inhibited signaling by the Rho family GTPases RAC1/CDC42/RhoA downstream of integrin. These findings indicate that CuB has been proven to mediate the reorganization and distribution of cytoskeletal proteins of breast cancer cells through RAC1/CDC42/RhoA signaling, which improves the mechanical properties of cell adhesion and deformation and consequently inhibits cell migration and invasion.     

Ethanol extract of Prunella vulgaris var. lilacina inhibits HMGB1 release by induction of heme oxygenase-1 in LPS-activated RAW 264.7 cells and CLP-induced septic mice

The ethanol extract of the flower of P. vulgaris var. lilacina (EEPV) has been used traditionally as an antiinflammatory agent in many countries. Inducers of heme oxygenase‐1 (HO‐1) reduce high mobility group box 1 (HMGB1), a late phase cytokine, in sepsis. Although EEPV has been used as an antiinflammatory agent, no report is available as to whether it modifies HMGB1 in sepsis due to HO‐1 induction. It was found that EEPV increased HO‐1 protein expression in RAW 264.7 cells, which was significantly inhibited by LY294002, but not PD98059, SB203580 or SP600125. In addition, EEPV activated NF‐E2‐related factor (Nrf2) to move from the cytosol to the nucleus, and EEPV‐induced HO‐1 and activation of ARE‐luciferase activity were significantly reduced by siNrf2 transfection and LY294002 but not SB203508. EEPV also significantly inhibited NF‐κB luciferase activity, and decreased both iNOS/NO and COX‐2/PGE₂ production in lipopolysaccharide (LPS)‐stimulated macrophages which was reversed by siHO‐1 RNA transfection. Importantly, EEPV inhibited HMGB1 release in LPS‐activated macrophages in a PI3K‐sensitive manner and reduced serum HMGB1 level and lung HMGB1 expression in cecal ligation and puncture (CLP)‐induced septic mice. It is concluded that EEPV induces HO‐1 expression through PI3K/Nrf2 signal pathways, which may be beneficial for the treatment of sepsis due to a reduction of HMGB1 release. Copyright © 2011 John Wiley & Sons, Ltd.     

Vaccinium angustifolium (lowbush blueberry) leaf extract increases extravillous trophoblast cell migration and invasion in vitro

Perturbations to extravillous trophoblast (EVT) cell migration and invasion are associated with the development of placenta‐mediated diseases. Phytochemicals found in the lowbush blueberry plant (Vaccinium angustifolium) have been shown to influence cell migration and invasion in models of tumorigenesis and noncancerous, healthy cells, however never in EVT cells. We hypothesized that the phenolic compounds present in V. angustifolium leaf extract promote trophoblast migration and invasion. Using the HTR‐8/SVneo human EVT cell line and Boyden chamber assays, the influence of V. angustifolium leaf extract (0 to 2 × 10⁴ ng/ml) on trophoblast cell migration (n = 4) and invasion (n = 4) was determined. Cellular proliferation and viability were assessed using immunoreactivity to Ki67 (n = 3) and trypan blue exclusion assays (n = 3), respectively. At 20 ng/ml, V. angustifolium leaf extract increased HTR‐8/SVneo cell migration and invasion (p < .01) and did not affect cell proliferation or viability. Chlorogenic acid was identified as a major phenolic compound of the leaf extract and the most active compound. Evidence from Western blot analysis (n = 3) suggests that the effects of the leaf extract and chlorogenic acid on trophoblast migration and invasion are mediated through an adenosine monophosphate‐activated protein (AMP) kinase‐dependent mechanism. Further investigations examining the potential therapeutic applications of this natural health product extract and its major chemical compounds in the context of placenta‐mediated diseases are warranted.     

异甘草素对结直肠癌细胞增殖、侵袭和迁移的影响

目的研究异甘草素对结直肠癌细胞增殖、侵袭和迁移的影响。方法 10、20、40、60、80、100、120μmol/L异甘草素处理HCT116、HT29细胞后,MTT法检测细胞毒性,流式细胞术检测HCT116细胞凋亡,Western blot法检测HCT116细胞中cleaved caspase-3、pJNK、JNK、pERK1/2、ERK1/2、pP38、P38蛋白表达,Transwell实验检测HCT116细胞侵袭和迁移。结果异甘草素对HCT116、HT29细胞的IC_(50)值分别为22.45、23.69μmol/L。与对照组比较,异甘草素组呈浓度依赖性地显著诱导HCT116细胞凋亡(P0.05),显著上调cleaved caspase-3蛋白表达(P0.05),显著抑制HCT116细胞侵袭和迁移(P0.05),显著下调pJNK/JNK、pERK1/2/ERK1/2、pP38/P38(P0.05)。结论异甘草素能抑制结直肠癌细胞增殖和侵袭,诱导细胞凋亡,其机制可能与抑制MAPK/ERK信号通路有关。     

基于miR-208a-3p探究萝卜硫素对肝癌细胞增殖、侵袭和迁移的影响

目的 探讨萝卜硫素对肝癌细胞增殖、侵袭和迁移的影响及其作用机制.方法 用10、20、40μmol/L萝卜硫素处理人正常肝细胞L02、肝癌HepG2细胞48h,通过CCK-8法筛选合适的萝卜硫素浓度用于后续实验;将HepG2细胞分为对照组和萝卜硫素(20μmol/L)组,CCK-8法检测细胞增殖;Transwell测定细...     

The antihypertensive and vasodilator effects of aqueous extract from Berberis vulgaris fruit on hypertensive rats

The aqueous extract from Berberis vulgaris fruit (B.V.) was tested to evaluate its antihypertensive effects on DOCA-induced hypertension in the rats. Hypertension was induced in male Sprague-Dawley rats (200-250 g) by DOCA-salt injection (20 mg/kg, twice weekly, for 5 weeks, s.c.) plus NaCl (1%) which was added to the animals' drinking water. Then 5 weeks later, the rats were anaesthetized with thiopental (30 mg/kg, i.p.) and the arterial blood pressure was measured. The mean arterial blood pressure and heart rate were 231 +/- 6.4 (mmHg) and 506 +/- 12 (beats/min), respectively. Administration of B.V. extracts significantly reduced the rat arterial blood pressure. In in vitro studies, rings of descending aorta were cut and mounted for isometric tension recording in an organ chamber containing Krebs solution. Mesenteric beds were also removed and perfused with Krebs solution. After 1 h of stabilization, preparations (aortic rings or mesenteric beds) were precontracted with phenylephrine (10(-5) M), then different concentrations of B.V. (0.4, 2 and 4 mg/mL) were added which caused a relaxation in these vessels. To investigate the mechanism of action of the extract, the tissues were incubated with either L-NAME (10(-5) M) or indomethacin (10(-5) M) for 20 min. In the aortic rings L-NAME pretreatment could only reduce the vasodilatory effects of a low concentration of B.V. (0.4 mg/mL), but indomethacin was without effect. In isolated perfused mesenteric beds preincubation with either L-NAME or indomethacin did not modify the vasodilator effects of the aqueous extract from B.V. fruit. The present results suggest that the antihypertensive and vasodilatory effects of B.V. fruit extract are mainly endothelial-independent and it may be used to treat hypertension, a status with endothelial dysfunction.     

消岩汤对肺癌A549细胞及肺癌干细胞迁移及侵袭的影响

目的探讨消岩汤对非小细胞肺癌转移的作用机制。方法通过划痕实验,观察消岩汤对肺腺癌A549细胞的迁移能力的影响;通过侵袭实验,观察消岩汤对肺癌干细胞(SP细胞)侵袭能力的影响。结果划痕实验中消岩汤组肺腺癌A549细胞在24、48、72 h的迁移距离明显短于各时刻对照组细胞(P0.05);侵袭实验中消岩汤干预下的SP细胞穿出Transwell小室的细胞数明显少于对照组(P0.05)。结论消岩汤可有效抑制肺腺癌A549细胞的迁移及SP细胞的侵袭。     

鸡血藤抗肿瘤有效部位诱导A549肺癌细胞非凋亡性程序化死亡的研究

目的：探讨鸡血藤抗肿瘤有效部位(SSCE)对人非小细胞肺癌A549细胞的杀伤特点和作用机制。方法：采用四氮唑溴盐(MTT)法、绘制生长曲线观察药物对A549细胞增殖的影响。采用流式细胞术分析药物对肿瘤细胞周期的影响。通过HE染色和激光共聚焦扫描荧光显微镜技术、AnnexinV-PI双标法，Caspase-3活性检测以及DNA琼脂糖凝胶电泳，观察药物对细胞凋亡的诱导作用。结果：SSCE对A549细胞有明显杀伤作用，24 h药效最强，IC₅₀为25.54 mg·L^-1。细胞周期分析显示，药物作用12 h，S期细胞增多，G₀-G₁和G₂-M期细胞减少，细胞周期变化在24 h后明显减弱，48 h恢复正常。形态学观察可见，药物作用短时间内(8～12 h)，细胞核发生皱缩扭曲，胞浆内出现较多空泡及颗粒状物，胞膜完整；随后细胞核固缩。细胞膜磷脂酰丝氨酸(PS)外翻在用药后12 h最显著，具有时间-剂量依赖关系。Caspase-3活性无明显升高，且与剂量成反比。DNA琼脂糖凝胶电泳未见片断化梯形条带。结论：SSCE对A549细胞具有直接杀伤作用，药效迅速，细胞周期受到干扰(S delay)，主要表现为非凋亡性程序化细胞死亡。     

HPLC法同时测定夏枯草中芦丁、金丝桃苷、槲皮素和山柰酚

目的 建立高效液相色谱法同时测定夏枯草中芦丁、金丝桃苷、槲皮素和山柰酚的分析方法.方法 用DiamonsilC18(4.6 mm× 150 mm,5μn)色谱柱分离,乙腈-0.1％磷酸进行梯度洗脱,体积流量1.0 mL/min,检测波长365 nm.结果 芦丁、金丝桃苷、槲皮素和山柰酚分别在1.88～37.60 μg/mL(r =0.999 9),5.28～ 39.60 μg/mL(r=0.999 9),1.32～26.40 μg/mL(r=0.999 9),0.168～13.44 μg/mL(r =0.999 9)范围内线性关系良好,平均回收率均大于97％.结论 本方法简便,准确,重复性好,可用于夏枯草的质量控制.     

The extract of Hibiscus syriacus inducing apoptosis by activating p53 and AIF in human lung cancer cells

Natural products including plants, microorganisms and marine life provide rich resources for anticancer drug discovery. The root bark of Hibiscus syriacus has been used as an antipyretic, anthelmintic and antifungal agent in Asia. The antiproliferative effects of H. syriacus on human lung cancer cells were evaluated with bio-assays. The apoptotic activity was detected by Hoechst 33342 DNA staining and annexin V staining. The expression of caspases, p53, apoptosis induced factor (AIF), Bcl-2 and Bax were evaluated with Western blotting. The in vivo anticancer activity was evaluated using A549-xenograft model. The acetone extract of H. syriacus (HS-AE) exhibited a better cytotoxic effect on lung cancer cells than its methanol extract (HS-ME) or water extract (HS-WE). The IC(50) values of HS-AE on A549 (adenocarcinoma), H209 (squamous cell carcinoma) or H661 (large cell carcinoma) lung cancer cells ranged from 14 to 22 microg/ml after 48 hours of treatment. After 48 hours of exposure, HS-AE (15 microg/ml) induced A549 cell apoptosis to 48 +/- 3.6% of the control. Using Western blotting, HS-AE appears to suppress the expression of p53 and AIF. The results of the in vivo study showed that HS-AE suppresses growth in A549 subcutaneous xenograft tumors. These results indicate that HS-AE exerts significant and dose-dependent antiproliferative effects on cancer cells in vitro and in vivo, which prompts us to further evaluate and elucidate the bioactive component(s) of H. syriacus.     

Ganoderma lucidum extract induces G1 cell cycle arrest, and apoptosis in human breast cancer cells

Ganoderma lucidum (Fr.) Karst is a traditional Chinese herb that has been widely used for centuries to treat various diseases including cancer. Herein, an ethanol-soluble and acidic component (ESAC), which mainly contains triterpenes, was prepared from G. lucidum and its anti-tumor effects in vitro were tested on human breast cancer cells. Our results showed that ESAC reduced the cell viability of MCF-7 and MDA-MB-231 cells in a concentration-dependent manner with IC(50) of about 100 μg/mL and 60 μg/mL, respectively. DNA damage was detected by Comet assay and the increased expression of γ-H2AX after ESAC treatment was determined in MCF-7 cells. Moreover, ESAC effectively mediated G1 cell cycle arrest in both concentration- and time-dependent manners and induced apoptosis as determined by Hoechst staining, DNA fragment assay and Western blot analysis in MCF-7 cells. In conclusion, ESAC exerts anti-proliferation effects by inducing DNA damage, G1 cell cycle arrest and apoptosis in human breast cancer cells.     

青蒿琥酯对人结肠癌HCT-8细胞侵袭能力影响及机制研究

目的研究青蒿琥酯对结肠癌细胞侵袭能力的影响以及对细胞间黏附分子-1(Intercellular adhesion molecule-1,ICAM-1)、血管内皮生长因子165(Vessel endothelium growth factor165,VEGF165)和血管生成素-2(Angiogenin-2,Ang-2)蛋白表达的影响。方法以人结肠癌细胞HCT-8为模型,采用软琼脂集落培养试验检测青蒿琥酯对肿瘤细胞侵袭抑制效应;Western blot法检测青蒿琥酯作用前后细胞ICAM-1、VEGF165和Ang-2蛋白表达变化。结果 3μmol/L青蒿琥酯即能有效抑制结肠癌HCT-8细胞诱导的软琼脂集落形成(P<0.05)。经青蒿琥酯处理后,人结肠癌HCT-8细胞ICAM-1、VEGF165和Ang-2蛋白表达呈剂量依赖性减少(3～12μmol/L,P<0.05)。结论青蒿琥酯可明显抑制结肠癌HCT-8细胞的侵袭,其作用机制可能与其下调ICAM-1、VEGF165和Ang-2蛋白表达有关。