4‘,5,7—三羟基异黄酮对Balb／c 3T3小鼠成纤维细胞恶性转化的抑制作用

目的４’,５,７－三羟基异黄酮与多种肿瘤的低发生风险呈正相关,此文旨在探讨这一化合物的癌化学预防作用。方法 用３－ＭＣＡ和ＴＰＡ诱导Ｂａｌｂ／ｃ３Ｔ３细胞,建立体外二阶段转化模型,用这一模型来观察ｇｅｎｉｓｔｅｉｎ对Ｂａｌｂ／ｃ３Ｔ３－Ａ３１小鼠成纤维细胞恶性转化的影响。结果 在对细胞没有明显细胞毒作用的浓度下,ｇｅｎｉｓｔｅｉｎ明显抑制细胞的恶性转化,机理研究表明,ｇｅｎｉｓｔｅｉｎ抑制ＴＰＡ刺     

Genistein抑制HT1080人纤维肉瘤细胞的体外侵袭作用

目的 观察ｇｅｎｉｓｔｅｉｎ对恶性肿瘤细胞侵袭及侵袭相关性质的影响,探讨蛋白酪氨酸激酶抑制剂用于肿瘤转移治疗的可能性。方法 ＨＴ１０８０细胞经２０μｍｏｌ／Ｌ或４０μｍｏｌ／Ｌ Ｇｅｎｉｓｔｅｉｎ处理３天后,分别用重建基底膜侵袭模型、粘附基质分析、Ｔｒａｎｓｗｅｌｌ小室趋化运动模型以及Ｎｏｒｔｈｅｒｎ ｂｌｏｔ分析来研究药物处理后细胞侵袭、粘附、运动以及基质蛋白酶基因表达的改变。结果 经ｇｅｎｉｓ     

genistein对人乳腺癌抑瘤效应及造血辐射防护作用的实验研究

大豆异黄酮ｇｅｎｉｓｔｅｉｎ具有抗氧化、抗增殖等多重抑瘤效应[1 ] 。我们在近期的体外研究中发现 ,ｇｅｎｉｓｔｅｉｎ放疗后给药和模拟放疗对肿瘤细胞有协同杀伤作用[2 ] 。为此 ,在探讨ｇｅｎｉｓｔｅｉｎ对动物照射防护作用基础上 ,我们通过裸鼠荷瘤模型 ,研究其在体内有无放疗协同效应 ,特别是在抑瘤的同时对造血的影响 ,为进一步拓展其临床应用打下基础。一、材料与方法1 实验动物与试剂 :ｇｅｎｉｓｔｅｉｎ (Ｓｉｇｍａ公司 )溶于二甲基亚砜 ,稀释后过滤备用 ;ＶＥＧＦ、ＥＧＦＲ组化试剂盒购自博士德公司。ＢＡＬＢ Ｃ ｎｕ ｎ…     

结缔组织生长因子转染对人乳腺癌细胞系生物学行为的影响

背景与目的:结缔组织生长因子(CTGF)是CCN家族成员之一,参与体内多种生理和病理生理过程。本研究探讨CTGF对人乳腺癌细胞生物学行为的影响。方法:构建含有CTGF开放读码框的真核表达质粒,通过脂质体介导的方法将正义质粒转染MCF-7细胞,反义质粒转染MDA-MB-231细胞。观察CTGF不同表达水平与乳腺癌细胞增殖、凋亡、周期分布和体外侵袭等生物学行为的关系。结果:转染正义质粒使MCF-7细胞CTGF表达上调,抑制细胞增殖,促进细胞凋亡,细胞发生G0G1期阻滞,体外侵袭能力下降。转染反义质粒使MDA-MB-231细胞CTGF表达下调,促进细胞增殖,减少细胞凋亡,增强细胞体外侵袭能力,但对细胞周期无影响。结论:CTGF具有抑制乳腺癌细胞生长和侵袭的功能。促进细胞凋亡是CTGF抑制乳腺癌细胞体外生长的机制之一。CTGF可能对细胞周期有一定的影响。     

Genistein抑制乳腺癌细胞生长的机制

目的 Ｇｅｎｉｓｔｅｉｎ抑制乳腺癌细胞生长的机制。方法 研究主要应用于Ｎｏｒｔｈｅｒｎ印迹杂交,Ｗｅｓｔｅｒｎ印迹杂交,质粒转染技术以及细胞凋亡检测法,探讨Ｇｅｎｉｓｔｅｉｎ抑制乳腺癌细胞生长的机制。结果 Ｇｅｎｉｓｔｅｉｎ可明显抑制不同ＥＲ状态和不同ｐ５３状态的乳腺癌细胞系的生长,同时显著诱导ｐ５３下游基因ｐ２１＾ＷＡＦ／ＣＩＰＩ蛋白和ｍＲＮＡ的表达,而导致ｐ２１＾ＷＡＦＩ／ＣＩＰＩ的表达增强主     

4—乙酰胺苯基维甲酸酯抑制肿瘤细胞侵袭重组基底膜及其作用机理

目的研究新维甲类化合物４－乙酰胺苯基维甲酸酯（４－ＡＰＲ）对Ｂ１６－Ｆ１０小鼠黑色素瘤细胞侵袭能力的抑制作用,探讨其作用机理。方法癌细胞侵袭能力用重组基底膜侵袭试验衡量;ＰＡＧＥ底物酶谱方法检测Ⅳ型胶原酶活性;斑点杂交检测ＣＮＥ－２Ｚ细胞ＴＩＭＰ－１ｍＲＮＡ表达;以细胞生长曲线评价药物对Ｂ１６－Ｆ１０细胞的生长抑制作用。结果在１０－５ｍｏｌ／Ｌ和１０－６ｍｏｌ／Ｌ时,４－ＡＰＲ对Ｂ１６－Ｆ１０小鼠黑色素瘤细胞侵袭重组基底膜的抑制率分别为５４．２％和４１．９％,对ＣＮＥ－２Ｚ细胞培养上清中的Ⅳ型胶原酶活性有降低作用。此外,４－ＡＰＲ可抑制Ｂ１６－Ｆ１０细胞与ＬＮ、ＦＮ和Ｍａｔｒｉｇｅｌ的粘附,诱导ＣＮＥ－２Ｚ细胞ＴＩＭＰ－１ｍＲＮＡ表达。结论４－ＡＰＲ抑制Ｂ１６－Ｆ１０细胞侵袭重组基底膜。４－ＡＰＲ的抗侵袭活性与抑制肿瘤细胞的粘附,降低肿瘤细胞培养上清中Ⅳ型胶原酶的活性和（或）诱导肿瘤细胞ＴＩＭＰ－１ｍＲＮＡ表达等有关。     

洛铂对ＭＣＦ-７细胞凋亡及ＭＴＤＨ基因表达的影响

目的　研究洛铂对人乳腺癌ＭＣＦ-７细胞凋亡的作用并探讨其对转移基因ＭＴＤＨ表达的影响。方法　ＭＴＴ比色法检测洛铂对ＭＣＦ-７细胞株的增殖抑制作用,流式细胞仪检测洛铂对ＭＣＦ-７细胞的凋亡率,ＲＴ-ＰＣＲ法检测洛铂对乳腺癌转移相关基因ＭＴＤＨ表达的影响。结果　５～４０ｍｇ／Ｌ洛铂对ＭＣＦ-７细胞增殖均具有明显抑制作用,呈药物浓度 时间依赖性（Ｐ＜０.０５）;５ｍｇ／Ｌ、１０ｍｇ／Ｌ、１５ｍｇ／Ｌ洛铂作用于ＭＣＦ-７细胞４８ｈ后凋亡率分别为１９.０８％、２４.２３％和６５.７１％。洛铂可以抑制ＭＴＤＨ基因表达,当浓度≥１０ｍｇ／Ｌ时,ＭＴＤＨｍＲＮＡ表达呈阴性。结论　洛铂可抑制ＭＣＦ-７细胞的增殖并可诱导其发生凋亡,并可能通过抑制ＭＴＤＨ基因的表达抑制乳腺癌细胞的侵袭转移。     

单克隆抗体PD4诱导人胃癌细胞系MGC-803的凋亡

目的凋亡（Ａｐｏｐｔｏｓｉｓ）是一种区别于坏死的细胞生理性死亡方式。肿瘤发生与凋亡的异常有十分密切的关系。单克隆抗体ＰＤ４能识别胃癌细胞表面的分子量为４００００的分子（Ｐ４０）。本研究是为了观察单抗ＰＤ４是否具有诱导胃癌细胞凋亡的作用。方法我们应用流式细胞术和末端脱氧核苷酸标记及ＤＮＡ电泳法观察单抗ＰＤ４对胃癌细胞ＭＧＣ－８０３增殖周期的影响以及对细胞杀伤作用的方式,并检测了ＭＧＣ－８０３细胞表面Ｆａｓ抗原的表达情况。结果ＰＤ４有阻滞细胞周期、通过诱发凋亡而抑制肿瘤细胞生长的作用,且ＭＧＣ－８０３细胞Ｆａｓ抗原表达为阴性。结论Ｐ４０是一个与细胞凋亡或增殖有关、且不同于Ｆａｓ的肿瘤相关抗原,他的分子克隆有助于抗体诱导细胞凋亡机制的阐明。     

ＲｈｏＡ小干扰ＲＮＡ抑制乳腺癌细胞株ＭＣＦ-７及其裸鼠皮下移植瘤的实验研究

目的　观察ＲｈｏＡ小干扰ＲＮＡ（ｓｉＲｈｏＡ）对乳腺癌细胞株ＭＣＦ-７增殖、迁移、周期和凋亡的影响以及对裸鼠移植瘤生长的影响。方法　ｓｉＲｈｏＡＬｉｐｏｆｅｃｔａｍｉｎｅ２０００介导下转染乳腺癌细胞ＭＣＦ-７,转染４８ｈ后,采用Ｗｅｓｔｅｒｎｂｌｏｔ技术检测ＲｈｏＡ蛋白的表达,ＭＴＴ实验检测ｓｉＲｈｏＡ转染细胞的增殖变化,损伤刮擦实验检测ｓｉＲｈｏＡ转染细胞的迁移能力,流式细胞仪检测ｓｉＲｈｏＡ转染细胞的周期和凋亡,裸鼠移植瘤实验检测ｓｉＲｈｏＡ对肿瘤生长的影响。结果　成功转染ｓｉＲｈｏＡ的肿瘤细胞,Ｗｅｓｔｅｒｎｂｌｏｔ显示ＲｈｏＡ蛋白表达在ＭＣＦ-７细胞中明显下降;ｓｉＲｈｏＡ对ＭＣＦ-７细胞的增殖、迁移均有显著的抑制作用并能促进肿瘤细胞凋亡及细胞周期中Ｓ期细胞减少,Ｇ１／Ｇ０期细胞增加;裸鼠移植瘤内重复注射ｓｉＲｈｏＡ后肿瘤生长明显减缓。结论　ｓｉＲｈｏＡ能够明显抑制ＲｈｏＡ基因在乳腺癌细胞ＭＣＦ-７中的表达,部分逆转ＭＣＦ-７的恶性生物学行为并抑制裸鼠移植瘤的生长。     

维甲酸抑制雌激素受体阳性乳腺癌细胞生长机制的研究

目的探讨雌激素受体的表达对维甲酸抑制乳腺癌细胞生长的影响和ＲＡＲα表达的变化。方法雌激素受体阴性的乳腺癌细胞株ＭＤＡ－ＭＢ－２３１细胞,采用分子生物学质粒转染技术,将ＥＲ阴性的乳腺癌细胞ＭＤＡ－ＭＤ－２３１转染成ＥＲ阳性细胞。结果发现ＥＲ转染后的细胞不仅ＲＡＲα的基础表达升高,而且ＲＡ能明显抑制该细胞的生长。同时还发现,无论是已确立的雌激素受体阳性的乳腺癌细胞株,还是雌激素受体转染后的细胞,雌激素都能明显刺激ＲＡＲα的表达。结论雌激素通过ＥＲ上调ＲＡＲα量的表达,在维甲酸对乳腺癌细胞生长的抑制中起着很重要的作用。     

Effects of Genistein on Proliferation and Cell Cycle of Salivary Adenoid Cystic Carcinoma Cells

Objective： To investigate the growth inhibiting effect of tyrosine protein kinase inhibitor, genistein, on human salivary adenoid cystic carcinoma SACC-83 cell line in vitro, and its effects on the expression of CyclinB1 protein and cell cycle. Methods： Effects of genistein on the growth of SACC-83 cells in vitro were measured with MTr assay. Cell cycle was detected with flow cytometry. The expressions of CyclinB1 and Cdkl proteins were measured with Western blot method, and the results of protein expression were quantitatively analyzed by FluorChem V2.0 software. The results were statistically analyzed by SPSSI 1.5 software. Results： Genistein inhibited the cell proliferation in a dose-dependant and time-dependant manner. The genistein-treated SACC-83 cells were arrested in the G2/M phase and had lower contents of CyclinB1 and Cdkl proteins compared with the control group. Conclusion： The growth inhibiting effect of genistein on SACC-83 cells may be associated with the regulations of genistein on the CyclinB1 and Cdk1 protein expressions and the cell cycle.     

Growth inhibition with reversible cell cycle arrest of carcinoma cells by flavone L86-8275.

BACKGROUND: Previous studies have shown that polyhydroxylated flavonoids such as quercetin and genistein can inhibit tumor cell growth in vitro, and preliminary in vivo studies of the flavone L86-8275 have shown growth inhibition of LX529 and A549 lung carcinomas. L86-8275 [(-)cis-5,7-dihydroxy-2-(2-chlorophenyl)-8[4-(3-hydroxy-1-methyl)- piperidinyl]-4H-1-benzopyran-4-one] is a flavone of novel structure. PURPOSE: The purpose of this study was to determine in vitro whether L86-8275 is a more potent inhibitor of growth in breast carcinoma and lung carcinoma cells than quercetin or genistein. METHODS: We studied the effects of L86-8275 on cell growth in seven breast carcinoma cell lines and five lung carcinoma cell lines. MDA468 breast carcinoma was then selected for further study. Cell proliferation was measured by a colorimetric dye reduction assay; synthesis of DNA, RNA, and protein by incorporation of the radioactive metabolic precursors thymidine, uridine, or leucine, respectively; adenosine triphosphate (ATP) content by a luciferase-mediated bioluminescence reaction; and cell cycle progression by the use of cell-synchronizing drugs (aphidicolin and nocodazole) and flow cytometry. RESULTS: L86-8275 was not cytotoxic to stationary-phase cells but reversibly inhibited the growth of cells in exponential growth phase. At concentrations of 25-160 nM, L86-8275 inhibited growth of human breast and lung carcinoma cell lines by 50%. MDA468 breast carcinoma cells were 60-fold and 400-fold more sensitive to L86-8275 than to quercetin and genistein, respectively. By 24 hours after addition of L86-8275, DNA synthesis in MDA468 cells was inhibited by greater than 95%, protein synthesis by 80%, and RNA synthesis by 40%-60%, under conditions that preserved cellular ATP levels at approximately 80%-90% of control values. When MDA468 cells released from aphidicolin-induced cell cycle arrest were exposed to 200 nM L86-8275, they completed the S phase but arrested in G2. When cells released from nocodazole-induced cell cycle arrest were exposed to 200 nM L86-8275, they completed mitosis but arrested in G1. CONCLUSIONS: L86-8275 is a potent, yet reversible, growth-inhibitory flavone that can selectively block cell cycle progression in vitro at more than one point in the cell cycle. IMPLICATIONS: These findings suggest that L86-8275 is a candidate for further preclinical development, as well as a model for the synthesis of other flavonoids that might potently delay cell cycle progression to achieve inhibition of tumor growth. Future studies need to address optimal schedules for antiproliferative activity in vivo and inhibition of clonogenic activity.     

Soy phytochemicals synergistically enhance the preventive effect of tamoxifen on the growth of estrogen-dependent human breast carcinoma in mice

The objective of this work was to determine the interactive effects between soy bioactive components and tamoxifen (TAM) on prevention of estrogen-dependent breast cancer (BRCA). We initially investigated the effects of soy isoflavone genistein and TAM on the growth and cell cycle progression of estrogen-dependent MCF-7 human BRCA cells, and on the expression of ERalpha, pS2 and EGFR genes in vitro. Genistein or TAM alone inhibited the growth of MCF-7 cells in part via G(1) phase arrest, but their combinations showed suggestive antagonistic effects. We further evaluated the effects of bioactive soy components and TAM on the growth inhibition of MCF-7 tumors in a clinically relevant breast tumor model. TAM and bioactive soy components, genistein and soy phytochemical concentrate (SPC), delayed the growth of MCF-7 tumors. The combination of TAM with genistein or SPC, especially at the lower dose of TAM, had synergistic effects on delaying the growth of MCF-7 tumors. Biomarker determination suggests that the combination of TAM and soy components may synergistically delay the growth of MCF-7 tumors via their combined effects on induction of tumor cell apoptosis and inhibition of tumor cell proliferation. In addition, genistein and TAM combination synergistically delayed the growth of breast tumor via decreased estrogen level and activity, and down-regulation of EGFR expression. The results from our studies suggest that further investigations may be warranted to determine if the combination of TAM and bioactive soy components may be used for prevention and/or treatment of estrogen-dependent BRCA.     

Genistein potentiates the radiation effect on prostate carcinoma cells.

We have shown previously that genistein, the major isoflavone in soybean, inhibited the growth of human prostate cancer cells in vitro by affecting the cell cycle and inducing apoptosis. To augment the effect of radiation for prostate carcinoma, we have now tested the combination of genistein with photon and neutron radiation on prostate carcinoma cells in vitro. The effects of photon or neutron radiation alone or genistein alone or both combined were evaluated on DNA synthesis, cell growth, and cell ability to form colonies. We found that neutrons were more effective than photons for the killing of prostate carcinoma cells in vitro, resulting in a relative biological effectiveness of 2.6 when compared with photons. Genistein at 15 microM caused a significant inhibition in DNA synthesis, cell growth, and colony formation in the range of 40-60% and potentiated the effect of low doses of 200-300 cGy photon or 100-150 cGy neutron radiation. The effect of the combined treatment was more pronounced than with genistein or radiation alone. Our data indicate that genistein combined with radiation inhibits DNA synthesis, resulting in inhibition of cell division and growth. Genistein can augment the effect of neutrons at doses approximately 2-fold lower than photon doses required to observe the same efficacy. These studies suggest a potential of combining genistein with radiation for the treatment of localized prostate carcinoma.     

The inhibitory effect of genistein on the growth and metastasis of a transplantable rat accessory sex gland carcinoma.

BACKGROUND: A cell line (K1) derived from a carcinogen-induced accessory sex gland carcinoma was used to examine the effects of the soybean extract, genistein, on tumor growth and metastasis. METHODS: Male Lobund-Wistar rats were injected s.c. with 20 million K1 cells; genistein (50 mg/kg BW) or the vehicle was administered s.c. every 12 h for 31 days. RESULTS: Genistein significantly inhibited tumor growth. Compared with controls, fewer genistein-treated rats developed invasive tumors (11% vs. 44%) or lymph node metastases (44% vs. 89%). No lung metastases were found in genistein-treated animals in contrast to controls (0% vs. 44%). Estrogenic side effects were precipitated in genistein-treated rats, including decreased accessory sex gland complex weight, increased pituitary weight, decreased testis weight, and decreased (BW). Serum testosterone was undetectable and serum prostate-specific acid phosphatase activity was 38% lower in genistein-treated rats compared with controls. Genistein concentrations in the solid tumors (2 nmol/g) were one-third those in blood. CONCLUSIONS: These data suggest that genistein may be a useful chemotherapeutic agent to inhibit the growth and metastasis of accessory sex gland cancers, such as those derived from the prostate.     

Effect of DNA Methyltransferase in Comparison to and in Combination with Histone Deacetylase Inhibitors on Hepatocellular Carcinoma HepG2 Cell Line

Background: DNA demethylating agents and histone deacetylase inhibitors can affect reactivation of geneexpression and apoptosis induction by DNA acetylation and demethylation. The aim of the present study was to analyzethe effects of DNA demethylating agent genistein (GE) and histone deacetylase inhibitor valproic acid VPA), aloneand combined, on hepatocellular carcinoma Hep G2 cell line. Methods: The cells were treated with various doses ofgenistein and valproic acid (alone and combined) and the MTT assay and flow cytometry were used to determine cellviability and apoptosis. Results: Genistein and valproic acid inhibited the growth of HepG 2 cells significantly. Resultof flow cytometry demonstrated that genistein and valproic acid (alone and combined) induce apoptosis significantly ina time‑dependent manner. Conclusions: Genistein and valproic acid can significantly inhibit proliferation and induceapoptosis in HepG2 cell line. The apoptotic effects of GE in combination with VPA were more significant that of eachcompound alone.