Effects of Genistein and Daidzein on the Proliferation, Invasion, Migration and Adhesion of Melanoma Cells

IntroductionGenistein and daidzein (Fig.1 ) ,two majorisoflavonoids in soybeans,were reported to playimportant roles in cancer prevention[1 ,2 ] .Previousstudies demonstrated that genistein and daidzeininhibited the growth of leukemia,breast,colonand prostate cancers[38] .Our earlier studies alsodemonstrated that daidzein enhanced the immunefunction in mice[9,1 0 ] . These functions madegenistein and daidzein promising candidates forcancer prevention. However,it is usually themetastatic disse…     

Effects of Genistein on Cell Cycle and Apoptosis of Two Murine Melanoma Cell Lines

The effects of genistein on several tumor cell lines were investigated to study the effects of gen- istein on cell growth, cell cycle, and apoptosis of two murine melanoma cell lines, B16 and K1735M2. These two closely related murine melanoma cell lines, however, have different responses to the genistein treat- ment. Genistein inhibits the growth of both the B16 and K1735M2 cell lines and arrests the growth at the G2/M phase. After treatment with 60 μmol/L genistein for 72 h, apoptosis and caspase activities were de- tected in B16 cells, while such effects were not found in K1735M2. Further tests showed that after genistein treatment the protein content and mRNA levels of p53 increased in B16, but remained the same in K1735M2. The protein content and mRNA levels of p21WAF1/CIP1 increased in both cell lines after treatment. The results show that genistein might induce apoptosis in B16 cells by damaging the DNA, inhibiting topoi- somerase II, increasing p53 expression, releasing cytochrome c from the mitochondria, and activating the caspases which will lead to apoptosis.     

合欢花水提物对大鼠脑胶质瘤C6细胞株的增殖抑制与凋亡诱导作用

采用MTT法检测合欢花水提物对C6细胞增殖活力的影响,流式细胞术检测该水提物对C6细胞的诱导凋亡作用.结果表明:合欢花水提物能抑制C6细胞的增殖,且具有剂量和时间依赖性,其在24 h、48 h、72 h时对C6细胞的半数抑制率(LD50)分别为:305 μg/mL、50 μg/mL、15 μg/mL.流式细胞术结果显示:200 tμg/mL、300μg/mL合欢花水提物均可诱导C6细胞发生早期凋亡,与对照组细胞相比,早期凋亡率分别由对照组的0.7％增加到了4.3％、14.3％.提示合欢花水提物抑制C6细胞增殖机理与诱导细胞凋亡途径有关.     

污泥中真菌Penicillium lanosum NT-121的细胞毒活性产物研究

以来源于污泥的抗肿瘤活性真菌P. lanosum NT-121活性代谢产物研究为例, 探索污染环境中生物资源高值利用(即药源微生物新菌株资源开发)的新途径。采用活性跟踪分离的实验策略, 利用液液萃取、柱层析、重结晶等方法从该活性菌株的发酵液乙酸乙酯萃取物总浸膏中分离出3个化合物, 利用现代波谱技术结合文献对照鉴定其化学结构分别为5α,8α-过氧-(22E,24R)-麦角甾-6,22-二烯-3β-醇(1), 槲皮素(2), terrein(3)。用MTT法测试化合物1, 2, 3抑制K562细胞的IC₅₀ 值分别为 45.8, 563.8和6.5 μg/ mL。这3个化合物虽为已知化合物, 但均为首次在P. lanosum中发现, 且化合物1和3对K562细胞有一定增殖抑制活性。     

BDE-209对人正常肝L-02细胞生长和凋亡的影响

采用Cell Counting Kit-8 (CCK-8)和Hoechst 33342/PI等, 分析十溴联苯醚(BDE-209)对人正常肝L-02细胞增殖和凋亡的影响。结果显示, 在不同浓度水平(0~70 μg/mL)的BDE-209中暴露24小时后, 细胞增殖抑制率随着BDE-209浓度的升高而升高。经SPSS-Probit-Logit法计算, 得出 BDE-209对人正常肝L-02细胞24小时的IC₅₀为127.08 ±24.93 μg/mL。在不同浓度水平(0~15 μg/mL)的BDE-209中暴露24小时后, 与对照组相比, 各剂量组细胞未出现明显的细胞形态变化, 仅在15 μg/mL剂量组观察到细胞减少的现象。凋亡染色后, 荧光显微镜检发现, 随着BDE-209浓度升高, 亮蓝、淡粉染色细胞数量增大, 人正常肝L-02细胞的凋亡率增高。与对照组相比, 各剂量组差异均具有统计学意义。分析结果表明, BDE-209可引起人正常肝L-02细胞增殖受到抑制, 并诱导细胞凋亡率增加。     

重组绿豆胰蛋白酶抑制剂片段对肠癌细胞SW480迁移的影响

探讨重组胰蛋白酶抑制剂活性片段(LysGP33)的抗肠癌效应.大肠杆菌原核表达LysGP33活性片段,GST亲和层析柱对表达蛋白进行纯化.MTT比色和细胞划痕方法检测LysGP33活性片段对SW480细胞增殖和迁移的影响.结果表明:10 μmol/L LysGP33活性片段对SW480细胞的增殖活性无明显影响,但能够有效地抑制SW480细胞的迁移,并呈现时间依赖效应.绿豆胰蛋白酶抑制剂抑制了肠癌细胞的迁移,在抗肿瘤浸润和转移中具有一定的应用前景.     

金雀异黄素对小鼠黑色素瘤B16BL6细胞增殖、迁移和黏附的影响

利用四甲基偶氮唑盐微量酶反应比色法、细胞划痕实验和细胞黏附实验等研究了金雀异黄素对小鼠黑色素瘤B16BL6细胞增殖、迁移以及与血管内皮细胞黏附的影响;利用免疫荧光实验观察了金雀异黄素对B16BL6细胞中微管和肌动蛋白分布的影响.结果表明,与对照组相比,金雀异黄素可以抑制B16BL6细胞的增殖、迁移和与血管内皮细胞的黏附,影响B16BL6细胞中肌动蛋白的组装,但不影响细胞中微管的组装.这些结果提示,金雀异黄素抑制B16BL6细胞的增殖、迁移和与血管内皮细胞的黏附可能是通过诱导细胞中肌动蛋白的重排来实现的.     

Cinobufacini-induced HeLa cell apoptosis enhanced by curcumin

When used in combination with certain chemotherapies, curcumin has been shown to increase apoptosis in several cancer cell lines. Here, we report the combined effects of curcumin and cinobufacini on human cervical carcinoma cells. The aim of this study was to examine whether curcumin could enhance apoptosis induced by cinobufacini. 3-(4,5-Dimethylthiazol-2-y1)-2,5- diphenytetrazolium bromide (MTT) assays revealed that the growth and proliferation of HeLa cells could be inhibited by 75% after a combined treatment of 25 μg/mL cinobufacini and 8 μg/mL curcumin. The combined treatment is 3 times more effective than treatment with 25 μg/mL cinobufacini alone. Annexin V-FITC/PI staining, morphological changes and immunofluorescence verified a significant enhancement in cinobufacini-induced apoptosis when cells were also exposed to curcumin. The data showed that the proportion of early apoptotic cells significantly increased from 15.43% in cells treated only with 25 μg/mL cinobufacini to 49.2% in cells treated with 25 μg/mL cinobufacini and 8 μg/mL curcumin. Moreover, compared with treatment of only 25 μg/mL cinobufacini, ROS production increased 1.7-fold, the intracellular free Ca 2+ concentration increased 1.5-fold, and the mitochon- drial membrane potential decreased by 20% in the combined treatment. Simultaneously, the atomic force microscopy (AFM) re- sults suggest that cells treated with a combination of cinobufacini and curcumin varied significantly in shape and ultrastructure. Collapsed cells with leaking cytoplasm, blebbing pores and emerging apoptotic bodies were prevalent. The nanoparticle size increased from 70 nm when the cells were treated with 25 μg/mL cinobufacini to 190 nm when the cells were treated with 25 μg/mL cinobufacini and 8 μg/mL curcumin. The size increase resulted in the cell membrane becoming considerably rough. These results can improve our understanding of combination treatments. Specifically, the combination of cinobufacini and curcumin may potentially find use as a novel cervical carcinoma treatment. Additionally, AFM is a powerful tool that can be used to explore cellular morphologies and ultrastructures.     

佛手叶挥发油对HeLa细胞形态与结构的影响

探讨了佛手叶挥发油对HeLa细胞形态与结构的影响.采用噻唑蓝(MTT)法检测细胞活力,倒置显微镜、荧光显微镜和电子显微镜观察细胞形态与结构的变化,免疫荧光结合激光共聚焦显微镜观察微管的分布.结果表明:佛手叶挥发油能够抑制HeLa癌细胞的增殖,呈剂量和时间依赖性;不同浓度的佛手叶挥发油处理HeLa癌细胞24 h后,低剂量(200μg/mL)佛手叶挥发油使HeLa细胞皱缩,染色质凝集,出现凋亡小体,微管解聚,表现为典型的凋亡特征;中、高剂量(400和800μg/mL)佛手叶挥发油使HeLa细胞膜裂解,内容物外泄,细胞碎片增多,表明细胞已坏死.     

Human Soluble TRAIL Protein Inducing Apoptosis in Osteosarcoma Cell

This study is to examine the effect of human recombinant soluble TRAIL （TNF-related apoptosis-inducing ligand） protein inducing apoptosis in MG-63 human osteosarcoma cells. The inhibitive rates of TRAIL to MG-63 cells were detected by MTT assay. The apoptosis induced by TRAIL in MG-63 human osteosarcoma cells was analyzed with FACS and TUNEL and the apoptotic bodies were observed by transmission electron microscope. MTT assay showed that the inhibitive rates of 500, 1 000, 2 000 and 4 000 ng/mL TRAIL for 24 h were 10.1%, 24.3%, 50.6% and 97.7% respectively. Flow cytometric analysis showed that after MG-63 cells were treated with 2 gg/mL TRAIL for 6 h, obvious apoptotic peak would immediately appear before diploid peak. Human soluble TRAIL protein can quickly kill MG-63 osteosarcoma cells selectively, and may have potential value for clinical treatment of osteosarcoma.     

大蒜素对人肺腺癌细胞SLC-89增殖的影响

目的:观察大蒜素对人肺腺癌细胞SLC-89增殖的影响。方法:不同浓度的大蒜素与SLC-89细胞共培养72 h,运用MTT法测定大蒜素的IC50。将IC50、IC50/2、IC50/4剂量的大蒜素与SLC-89细胞共培养72 h后,运用端粒重复序列扩增-酶联免疫吸附法分别测定细胞端粒酶活性,运用流式细胞技术测定细胞PCNA的表达率。加入等量生理盐水共培养作为对照组。结果:大蒜素能抑制SLC-89细胞的增殖,其IC50为79.8μg/mL,IC50和IC50/2剂量的大蒜素能显著抑制SLC-89细胞端粒酶的活性,IC50、IC50/2、IC50/4剂量的大蒜素都能显著抑制SLC-89细胞PCNA的表达。结论:大蒜素能抑制SLC-89细胞的增殖,其PCNA的表达比端粒酶活性更容易受到下调。     

喙荚云实的化学成分研究

对喙荚云实Caesalpinia minax枝和叶的化学成分进行了研究.利用现代多种色谱技术从喙荚云实枝和叶的乙醇提取物中分离得到7个化合物,利用波谱技术鉴定了它们的结构,分别为S(+)-dehydrovomifoliol(1)、loliolide(2)、vomifoliol(3)、taraxerol(4)、3,13-dihydroxy-12-oleananone(5)、glutinol(6)、3β-hydroxysitost-5-en-7-one(7),化合物类型主要为倍半萜和三萜,所有化合物均为首次从喙荚云实中得到.     

五种中草药提取液对细菌作用效果初探

选取银杏、厚朴、野菊花、苦参、石菖蒲五种中草药,利用95%酒精提取其活性成分,进行抑菌、杀菌活性测定.结果表明,在供试质量浓度0.05g/mL时,此五种中草药对大肠杆菌、枯草杆菌、金黄色葡萄球菌的抑菌率均在86%以上;在供试质量浓度0.01g/mL时,作用时间为5min,五种中草药对大肠杆菌、金黄色葡萄球菌的杀菌率均在94.5%以上,杀菌效果显著.不同中草药对同一细菌的抑制作用有一定差异,同一中草药对不同细菌的抑制作用亦存在着较大差异.中草药的杀菌率随着浓度增大、着用时间延长,杀菌率随之增大.但五种中草药对枯草杆菌均无杀菌效果,其原因有待进一步探讨.     

苦楝叶乙醇浸提物对几种作物种子萌发及幼苗生长的影响

以苦楝（Melia azedarach）叶乙醇浸提物处理大豆（Glycine mac）、水稻（Oryza stativa）、小麦（Triticum aes-tivum）及玉米（Zeamays）种子,观察浸提物对几种作物种子萌发及幼苗生长的影响。结果表明,在62.5～1000μg/mL浓度范围内,浸提物对几种作物种子的萌发及幼苗生长具有一定的抑制作用,但对水稻和小麦的抑制作用较强,对玉米和大豆的抑制作用稍弱,在1000μg/mL浓度时,浸提物可完全抑制水稻和小麦种子萌发及幼苗的生长,而大豆和玉米的萌发率仍超过70%,对大豆幼苗的苗高和根长,玉米幼苗的苗高抑制作用均低于50%。     

Inhibition Mechanism of Emodin on Rabbit Vascular Smooth Muscle Cells Proliferation

ＩｎｔｒｏｄｕｃｔｉｏｎＥｍｏｄｉｎｉｓｏｎｅｏｆｔｈｅａｎｔｈｒａｑｕｉｎｏｎｅｃｏｍｐｏｕｎｄｓ[1] ,ｗｈｉｃｈｅｘｉｓｔｓｉｎｍａｎｙｍｅｄｉｃａｌｐｌａｎｔｓｓｕｃｈａｓｒａｄｉｘ ｐｏｌｙｇｏｎｉｍｕｌｔｉｆｌｏｒｉ,ｒｈｉｚｏｍａｐｏｌｙｇｏｎｉ,ｅｔｃ .Ｅｍｏｄｉｎｉｓａｎｉｍｐｏｒｔａｎｔｃｏｍｐｏｕｎｄｉｎｍｅｄｉｃｉｎｅ .Ｉｔｈａｓｍａｎｙｂｉｏｌｏｇｉｃａｌａｃｔｉｖｉｔｉｅｓｓｕｃｈａｓａｎｔｉ ｉｎｆｅｃｔｉｏｎ ,ｄｅｃｒｅａｓｉｎｇｂｌｏｏｄｌｉｐｉｄｓｃｏｎｔｅｎｔ ,ｅ…     

桑黄粗多糖对肿瘤细胞增殖及转移相关能力的抑制作用

研究了桑黄粗多糖（PL）对高转移卵巢癌细胞（HO-8910PM）和乳腺癌细胞（Bcap-37）增殖和转移相关能力的影响及其作用的机制.研究结果表明,桑黄粗多糖能有效抑制裸鼠移植瘤的生长,抑制率为65.37%;对离体培养的HO-8910PM和Bcap-37细胞的抑制率分别达到15.53%和23.81%;500 ind/ml PL作用于HO-8910PM细胞96 h可诱导其凋亡,通过细胞周期G0/G1期阻滞而抑制细胞增殖和诱导细胞凋亡;PL能显著抑制HO-8910PM的黏附、侵袭和迁移等肿瘤转移相关能力.桑黄粗多糖具有抑制肿瘤细胞增殖及肿瘤转移的作用.     

玉竹中高异黄酮类化合物生物活性研究

采用四唑盐(MTT)比色法、白细胞弹性蛋白酶抑制剂筛选模型及二氢乳清酸脱氢酶抑制剂筛选模型对从玉竹中分离出的4个高异黄酮类化合物进行生物活性评价,探讨化合物结构与其活性之间的构效关系.高异黄酮类化合物1～4对肿瘤细胞K562,A549和HCT-15具有细胞毒性,IC50为7～35μg/mL;化合物1～4对人白细胞弹性蛋白酶有抑制作用,IC50分别为13.1,70.4,13.8,55.2μg/mL;化合物2,4对二氢乳清酸脱氢酶有一定的抑制作用,IC50分别为10.0,11.1μg/mL.高异黄酮类化合物的生物活性与其取代基的类型及位置有关.     

蒜头果果仁中7种金属元素含量的测定

采用湿法硝酸—高氯酸对蒜头果果仁进行了消解前处理,用电感耦合等离子体原子发射光谱法(ICP-AES)测定了蒜头果果仁中Al、Ca、Fe、K、Mg、Mn、Zn 7种金属元素的含量。结果表明,7种元素含量分别为18.65μg/g,192.6μg/g,9.946μg/g,12.12 mg/g,2.259 mg/g,25.55μg/g和23.46μg/g。元素的检出限为0.001 2～0.169 8μg/mL,RSD=1.04%～7.58%(n=5),回收率为97.49%～109.6%。建立了蒜头果果仁中7种金属元素含量测定方法。该方法简便、快速,测定结果准确可靠。     

中空介孔硅球对血管内皮细胞的细胞毒性

通过细胞增殖实验和乳酸脱氢酶释放实验,流式细胞术,透射电镜及电感耦合等离子体发射光谱仪检测等方法,证实HMSN在浓度超过62.5 μg/mL时,会对人脐静脉血管内皮细胞产生毒性,半抑制浓度约为150 μg/mL.高浓度下HMSN导致细胞增殖抑制和乳酸脱氢酶释放,使细胞周期阻滞在G1期,并最终诱导细胞坏死.同时发现HMS...     

微量进样-火焰原子吸收分光光度法测定红细胞以及其它组织中的铅

在国产GGX-600型原子吸收分光光度计上采用微量进样技术,用火焰原子吸收光谱法测定了小白鼠红细胞、血清、肝、肾、脾、肺、心脏和大脑中铅的含量。实验结果表明该方法铅在0．050～1．000μg／mL呈良好线性关系,r=0．9997,方法平均回收率97．4％,RSD=4．2％,特征浓度为0．0916μg/mL,检出限为0．0500μg／mL。方法操作简单,重现性好,结果准确且待测样品用量少。