gamma rays. The effect of cucurbitacin B on cell-survival following irradiation was evaluated by colony-forming assay. Cell cycle distributions were investigated using flow cytometry. Real-time PCR and western blots were performed to investigate the expression of cell cycle checkpoints. Results. Cucurbitacin B inhibited breast cancer cell proliferation in a dose-dependent manner. Only MDA-MB-231 and MCF7:5C cells but not SKBR-3 cells were radiosensitized by cucurbitacin B. Flow cytometric analysis for DNA content indicated that cucurbitacin B resulted in G2/M arrest in MDA-MB-231 and MCF7:5C but not SKBR-3 cells. Moreover, Real-time PCR and western blot analysis demonstrated upregulated p21 expression before irradiation, a likely cause of the cell cycle arrest. Conclusion. Taken together, these findings suggest that cucurbitacin B causes radiosensitization of some breast cancer cells, and that cucurbitacin B induced G2/M arrest is an important mechanism. Therefore, combinations of cucurbitacin B with radiotherapy may be appropriate for experimental breast cancer treatment.">
Cucurbitacin B导致增加人类乳腺癌细胞的辐射敏感性通过G2 / M细胞周期阻滞</t我tle>
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<h1 class="articleHeader__title">Cucurbitacin B导致增加人类乳腺癌细胞的辐射敏感性通过G2 / M细胞周期阻滞</h1>
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<h4 class="header" id="abstract">文摘</h4>
<p><i>目的</我>。探索cucurbitacin B辐射的影响生存的人类乳腺癌细胞和细胞机制的阐明放射线增减。<我>材料和方法</我>。人类乳腺癌细胞系之前接受cucurbitacin B照射的清廉Gy<年代vg height="13.9875" id="M1" style="vertical-align:-0.17554pt;width:33.662498px;" version="1.1" viewbox="0 0 33.662498 13.9875" width="33.662498" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,13.9875)">
<g transform="translate(72,-60.81)">
<text transform="matrix(1,0,0,-1,-71.95,66.03)">
<tspan style="font-size: 8.75px; " x="0" y="0">
1</t年代pan>
<tspan style="font-size: 8.75px; " x="4.3759999" y="0">
3</t年代pan>
<tspan style="font-size: 8.75px; " x="8.7519999" y="0">
7</t年代pan>
</text>
<text transform="matrix(1,0,0,-1,-58.32,61.03)">
<tspan style="font-size: 12.50px; " x="0" y="0">
C</t年代pan>
<tspan style="font-size: 12.50px; " x="8.3395014" y="0">
年代</t年代pan>
</text>
</g>
</g>
</svg>伽马射线。cucurbitacin B在照射后细胞存活的影响被克隆形成试验评估。使用流式细胞仪细胞周期分布进行调查。实时PCR和西方印迹进行调查的表达细胞周期检查点。<我>结果。</我>Cucurbitacin B的剂量依赖性地抑制乳腺癌细胞增殖。只有mda - mb - 231和MCF7:5C细胞但不是SKBR-3细胞被cucurbitacin radiosensitized B流仪分析DNA内容表明cucurbitacin B导致G2 / M在mda - mb - 231和被捕MCF7:5C但不是SKBR-3细胞。此外,实时聚合酶链反应和免疫印迹分析表明辐照前调节p21表达,细胞周期阻滞的可能原因。<我>结论</我>。总的来说,这些发现表明,cucurbitacin B导致放射线增减一些乳腺癌细胞,B, cucurbitacin诱导G2 / M逮捕是一个重要的机制。cucurbitacin B的因此,组合与放疗可能适合实验乳腺癌治疗。</p>
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<h4 id="introduction">1。介绍</h4>
<p>乳腺癌是女性最常见的原因女性癌症和癌症死亡的主要原因,在美国和世界其他地区<一个href="#B7">1</一个>,<一个href="#B12">2</一个>]。在过去的几十年里,乳腺癌的发病率一直在增加在经济发达的国家<一个href="#B14">3</一个>]。人类表皮生长因子受体2 (Her2)和雌激素受体(ER)的开发和发展发挥了关键作用乳腺癌。大约80%的乳腺癌是激素受体阳性和表达雌激素受体(<一个href="#B15">4</一个>]。大约20%的乳腺癌也不表达雌激素受体和Her2 [<一个href="#B1">5</一个>,<一个href="#B13">6</一个>]。因此,乳腺癌为ER和Her2阴性不应对荷尔蒙疗法。目前治疗乳腺癌的治疗可能导致耐药性和毒性。越来越多的兴趣使用草药帮助女性健康的维护;有趣的是使用草药的妇女的卫生保健。植物含有各种各样的化学物质有强大的生物效应,包括抗癌活性。自然cucurbitacins高含氧,四环三萜烯含有cucurbitane核骨架和主要是发现在家庭葫芦科植物,成员长期以来一直用于东方药物,因为他们表现出的广泛生物活性的植物和动物。在各种cucurbitacins中,最丰富的是cucurbitacin B cucurbitacin B(计划<一个href="//www.newsama.com/journals/jo/2012/sch1/" target="_blank">1</一个>)从泰国草药中提取<我>Trichosanthes cucumerina</我>l .已被证明有抗癌,抗菌,抗炎活动(<一个href="#B25">7</一个>,<一个href="#B11">8</一个>]。一些研究报道,cucurbitacin B和其亲属抑制人类恶性肿瘤细胞的生长<我>在体外</我>和<我>在活的有机体内</我>包括乳腺癌[<一个href="#B26">9</一个>),头颈鳞状细胞癌(<一个href="#B19">10</一个>[],胰腺癌<一个href="#B10">11</一个>),肝细胞癌(<一个href="#B2">12</一个>),骨肉瘤(<一个href="#B17">13</一个>),和骨髓性白血病<一个href="#B9">14</一个>]。我们以前的报告表明,cucurbitacin B产生抗癌作用通过抑制端粒酶通过下调了<我>hTERT</我>和<我>原癌基因</我>表达和细胞周期的逮捕G<年代ub>2</年代ub>/ M期乳腺癌细胞(<一个href="#B5">15</一个>]。有研究表明,细胞对辐射最敏感的G<年代ub>2</年代ub>/ M和S期最耐[<一个href="#B20">16</一个>]。例如,同步中国仓鼠细胞最敏感的辐照在有丝分裂和G<年代ub>2</年代ub>阶段和不敏感的G<年代ub>1</年代ub>而后者过去S期(<一个href="#B24">17</一个>,<一个href="#B23">18</一个>]。药物,包括多烯紫杉醇,逮捕细胞细胞G<年代ub>2</年代ub>/ M期细胞周期已被证明为辐射敏化剂的代理(<一个href="#B8">19</一个>]。本研究的目的是确定的机体潜力cucurbitacin B在人类乳腺癌细胞和阐明细胞机制的放射线增减。在目前的研究中,我们证明了B cucurbitacin糖分会让人类乳腺癌细胞对辐射诱导他们积聚在G<年代ub>2</年代ub>/ M细胞周期的阶段。</p>
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<span class="caption-text">方案1</年代pan>
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<td>cucurbitacin B的结构。</td>
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<h4 id="materials-and-methods">2。材料和方法</h4>
<h5 id="sec2.1">2.1。细胞系和药物治疗</h5>
<p>人类乳腺癌细胞系(SKBR−3 (ER−/ Her2 +), mda - mb - 231 (ER / Her2−−)和hormone-independent MCF7:5C (ER / Her2−−)是培养在37°C下5%的股份有限公司<年代ub>2</年代ub>的气氛。SKBR-3乳腺癌细胞维持本人的5中。mda - mb - 231和MCF7:5C DMEM / F12培养基中维护。所有媒体都补充了10%胎牛血清和1%青霉素和链霉素。</p>
<h5 id="sec2.2">2.2。药物和放射治疗</h5>
<p>Cucurbitacin B被教授身份验证Apichart suksamrarn从理学院,Ramkhamhaeng大学,泰国曼谷。这种化合物溶解在10%二甲亚砜(DMSO)和稀释DMEM / F12培养基和真品是5中使用之前所需的浓度。铯机用于细胞辐射剂量范围从0到10 Gy。</p>
<h5 id="sec2.3">2.3。单独生存分析</h5>
<p>细胞被播种在100毫米的文化板块和对待cucurbitacin B的浓度表示辐射暴露前48小时。曝光后,细胞被使胰蛋白酶化和种子密度差的基础上在一个60毫米培养板5毫升的媒介。板块在37°C下5%孵化有限公司<年代ub>2</年代ub>氛围的程度。这些细胞被固定在乙醇和结晶紫染色。包含50多个细胞被算作殖民地幸存者。幸存的分数的计算是通过规范化的电镀效率适当的对照组。</p>
<h5 id="sec2.4">2.4。细胞周期分析</h5>
<p>细胞周期分析,细胞治疗cucurbitacin B在不同浓度48小时和收获。这些细胞被使胰蛋白酶化和resuspend 1毫升DPBS。一百万细胞离心和悬浮在0.5毫升Krishan试剂(0.1%柠檬酸钠,0.03% NP-40, 0.05毫克/毫升π,0.02毫克/毫升核糖核酸酶A)之前的分析。染色细胞DNA受到内容/细胞周期分析使用一个LSR流式细胞分析仪。</p>
<h5 id="sec2.5">2.5。细胞凋亡分析</h5>
<p>细胞凋亡的膜联蛋白V-FITC凋亡检测设备(BD生物科学、贝德福德,MA)被用来评估膜联蛋白V-positive细胞。简单、新鲜细胞准备孵化1 x膜联蛋白绑定缓冲和膜联蛋白V-FITC - (2.5<我>μ</我>g / mL)共轭主要抗体15分钟在冰上。孵化后,propidium碘(π;10<我>μ</我>g / mL)添加到悬浮细胞被流式细胞术分析使用一个LSR流式细胞分析仪。</p>
<h5 id="sec2.6">2.6。量化的<我>p21</我>信使核糖核酸</h5>
<p>细胞(<年代vg height="14.5" id="M2" style="vertical-align:-0.3135pt;width:46.612499px;" version="1.1" viewbox="0 0 46.612499 14.5" width="46.612499" xmlns="http://www.w3.org/2000/svg">
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</svg>细胞/)被播种到6-well板和处理各种浓度的cucurbitacin B 48小时。从每个细胞总RNA分离线使用试剂盒RNeasy迷你包(试剂盒,瓦伦西亚,CA)。两个微克的总RNA reverse-transcribed用随机引物根据生产的协议使用高容量cDNA逆转录工具包(应用生物系统公司,培育城市,CA)。实时PCR进行使用快速SYBR绿色主人混合(应用生物系统公司)与应用生物系统公司7500快速实时PCR系统(应用生物系统公司)。PCR引物如下:sense5′-TGAGCCGCGACTGTGATG-3′和anti-sense5′-GTCTCGGTGACAAAGTCGAAGTT-3′<我>p21和</我>sense5′-GAAGGTGAAGGTCGGAGTC-3′和anti-sense5′-GAAGATGGTGATGGGATTTC-3′<我>GAPDH</我>。的相对比例<我>p21</我>然后使用公式计算:<年代vg height="14.0625" id="M3" style="vertical-align:-0.0pt;width:288.20001px;" version="1.1" viewbox="0 0 288.20001 14.0625" width="288.20001" xmlns="http://www.w3.org/2000/svg">
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<h5 id="sec2.7">2.7。免疫印迹分析</h5>
<p>cucurbitacin B治疗后,与100年收集细胞颗粒和细胞溶解<我>μ</我>L•瑞帕细胞裂解缓冲(50毫米三pH值8.0,150毫米氯化钠,0.1% SDS, 0.5%钠脱氧胆酸盐,tx - 100) 1%氟化钠+ 1毫米,10毫米衣饰归宿<年代ub>4</年代ub>、10毫米PMSF和1/100蛋白酶抑制剂鸡尾酒(σ)。总蛋白测定使用Bio-Rad蛋白质测定(生命科学、大力神、钙)。等量的蛋白质分离在硝化纤维膜12.5% SDS-Polyacrylamide凝胶和electrotransferred p21和对待反(圣克鲁斯生物技术公司,圣克鲁斯,CA)过夜。平等蛋白质加载被确认在所有使用beta-tubulin抗体免疫印迹(杂种细胞发育研究银行,爱荷华大学爱荷华市,IA)稀释500。山羊anti-rabbit免疫球蛋白(BD转导实验室、圣地亚哥、CA)被用作二次抗体对所有主要抗体。乐队是可视化与发射极耦合逻辑+化学发光试剂(皮尔斯,罗克福德,IL)在7000年佛罗里达州台风。</p>
<h5 id="sec2.8">2.8。统计分析</h5>
<p>所有实验进行至少三次。统计分析中使用单向方差分析比较影响执行控制(没有cucurbitacin)和细胞治疗。<我>P</我>值< 0.05被认为是具有统计学意义。</p>
<h4 id="results">3所示。结果</h4>
<h5 id="sec3.1">3.1。Cucurbitacin B诱发单独抑制乳腺癌细胞</h5>
<p>cucurbitacin B的抑制作用在人类乳腺癌细胞集落形成被单独分析评估。细胞被孵化cucurbitacin B仅48小时,然后允许殖民地的新媒介形式。幸存的分数作为药物浓度的函数图所示<一个href="//www.newsama.com/journals/jo/2012/fig1/" target="_blank">1</一个>。平均50% (IC<年代ub>50</年代ub>)抑制浓度三个细胞克隆细胞细胞死亡为3.2,2.4和1.9<我>μ</我>MCF7:5C M, mda - mb - 231和SKBR-3分别。结果平均为每个细胞株从三个独立的实验。在单独使用化验SKBR-3最敏感的细胞cucurbitacin B在同等条件下其他细胞。</p>
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<td>cucurbitacin B的抑制性影响乳腺癌细胞的集落形成。MCF7:5C、mda - mb - 231和SKBR-3 cucurbitacin B的浓度表示治疗,疗程48小时。孵化后,细胞被播种密度差的基础上在一个60毫米和5毫升新鲜培养基培养板。在播种后14天,殖民地固定和染色结晶紫为0.1%。结果显示三个独立实验的平均值。<年代vg height="15.4375" id="M4" style="vertical-align:-3.13504pt" version="1.1" viewbox="0 0 65.099998 15.4375" width="65.099998" xmlns="http://www.w3.org/2000/svg">
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<h5 id="sec3.2">3.2。细胞周期</h5>
<p>影响MCF7:5C cucurbitacin B细胞周期进展,mda - mb - 231和SKBR-3细胞分析根据每个阶段的DNA含量的原理,细胞周期。细胞治疗cucurbitacin B 48小时,并通过流式细胞仪DNA含量进行了分析。MCF7:5C和mda - mb - 231细胞治疗被捕后G<年代ub>2</年代ub>/ M期细胞周期G1细胞数量的减少和S期细胞周期,就像观察到在一些癌症细胞系。然而,在对比的效果cucurbitacin B MCF7:5C和mda - mb - 231细胞,cucurbitacin G B没有贡献<年代ub>2</年代ub>(图/ M阶段逮捕SKBR-3细胞<一个href="//www.newsama.com/journals/jo/2012/fig2/" target="_blank">2</一个>)。</p>
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<td>cucurbitacin B对乳腺癌细胞的细胞周期进程。MCF7:5C、mda - mb - 231和SKBR-3 cucurbitacin B治疗,疗程48小时,然后沾propidium碘(PI)和受流仪分析。DNA直方图显示代表三个独立的实验。堵塞在G<年代ub>2</年代ub>/ M和观察凋亡诱导(SubG<年代ub>1</年代ub>)。</td>
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<h5 id="sec3.3">3.3。细胞凋亡Cucurbitacin B对乳腺癌细胞的影响</h5>
<p>cucurbitacin B细胞凋亡效应评估通过膜联蛋白V-FITC和propidium碘染色。这个试验表明带负电荷的磷脂磷脂酰丝氨酸的内表面上发现细胞的质膜是trans-located在凋亡细胞表面。48小时孵化后0<我>μ</我>2.5米,<我>μ</我>5米,<我>μ</我>M cucurbitacin B细胞被染色,进行二元流仪分析。如图<一个href="//www.newsama.com/journals/jo/2012/fig3/" target="_blank">3</一个>,未经处理的细胞没有任何明显的细胞凋亡,而成为细胞凋亡与cucurbitacin B治疗表示浓度在所有细胞。</p>
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<td>乳腺癌细胞诱导的细胞死亡cucurbitacin B . MCF7:5C mda - mb - 231, SKBR-3孵化cucurbitacin B 48小时和分析了细胞凋亡染色磷脂酰丝氨酸易位FITC-Annexin诉膜联蛋白V染色的表示<年代vg height="7.1624999" id="M6" style="vertical-align:-0.11285pt" version="1.1" viewbox="0 0 8.7250004 7.1624999" width="8.7250004" xmlns="http://www.w3.org/2000/svg">
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<h6 id="sec3.3.1">3.3.1。<我>p21</我>信使rna表达</h6>
<p>确定cucurbitacin B的影响<我>p21</我>mRNA表达,所有三个细胞株孵化cucurbitacin B为48小时,与未经处理的细胞。暴露MCF7:5C、mda - mb - 231和SKBR-3细胞株至2.5<我>μ</我>米和5<我>μ</我>M cucurbitacin B导致进步的增加<我>p21</我>信使rna水平。p21 SKBR-3显示最高感应的mRNA表达cucurbitacin B治疗后。表达式<我>p21</我>信使rna在SKBR-3增加了20倍与未经处理的细胞相比,MCF7:5C和mda - mb - 231如图增加3 - 4倍<一个href="//www.newsama.com/journals/jo/2012/fig4/" target="_blank">4(一)</一个>。实时PCR产品被应用在0.8%琼脂糖凝胶含有溴化乙锭(EtBr)审查,PCR反应是特定的,cucurbitacin B诱导的基因表达<我>p21</我>。</p>
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<td>cucurbitacin B的影响<我>p21</我>基因的表达。MCF7:5C、mda - mb - 231和SKBR-3孵化了48小时的指定cucurbitacin B的浓度和RNA提取实时PCR量化的表达水平<我>p21</我>。的相对表达水平<我>p21</我>信使rna浓度表示。结果显示三个独立实验的平均值。<年代vg height="15.4375" id="M10" style="vertical-align:-3.13504pt" version="1.1" viewbox="0 0 65.099998 15.4375" width="65.099998" xmlns="http://www.w3.org/2000/svg">
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<h6 id="sec3.3.2">3.3.2。Upregulation Cucurbitacin p21蛋白的B</h6>
<p>我们检查的影响cucurbitacin B细胞周期调控蛋白的表达,免疫印迹分析。B细胞被孵化的表示浓度cucurbitacin 48小时,提取总蛋白免疫印迹分析。如图<一个href="//www.newsama.com/journals/jo/2012/fig5/" target="_blank">5</一个>,p21蛋白表达的细胞周期蛋白依赖性激酶抑制剂cucurbitacin B治疗后显著增加在所有研究细胞。SKBR-3细胞,显示最高的mRNA积累应对cucurbitacin B,也显示出最大的感应p21蛋白;然而,这并不一定与G<年代ub>2</年代ub>cucurbitacin B / M逮捕或辐射敏感。</p>
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<td>影响cucurbitacin B p21蛋白表达在乳腺癌细胞。(a)与cucurbitacin B细胞治疗48小时,然后提取总蛋白进行免疫印迹分析<年代vg height="13.55" id="M12" style="vertical-align:-2.29482pt" version="1.1" viewbox="0 0 23.4 13.55" width="23.4" xmlns="http://www.w3.org/2000/svg">
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<h5 id="sec3.4">3.4。幼崽的Radiopotentiating效应在乳腺癌细胞</h5>
<p>确定cucurbitacin B敏感人类乳腺癌细胞电离辐射,所有三个细胞株服用5<我>μ</我>M cucurbitacin B的辐照后48小时<年代up>137年</年代up>Cs在剂量辐照器主Gy不等。细胞被允许殖民地的新媒介形式。所有细胞的电镀效率是60 - 80%。图<一个href="//www.newsama.com/journals/jo/2012/fig6/" target="_blank">6</一个>生存曲线显示了辐射来自单独分析的三个细胞株辐照后48小时孵化与cucurbitacin B .后者MCF7存活曲线的斜率:C和mda - mb - 231细胞对辐射和cucurbitacin B治疗大于辐射,特别是在6和8 Gy辐射剂量,表明cucurbitacin B治疗增强辐射的影响在两个细胞系,G<年代ub>2</年代ub>观察/ M逮捕发生。然而,SKBR-3没有显示增强辐射的影响,与细胞周期分布如图一致<一个href="//www.newsama.com/journals/jo/2012/fig2/" target="_blank">2</一个>和G的缺失<年代ub>2</年代ub>在SKBR-3细胞/ M逮捕。</p>
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<td>单独使用治疗后乳腺癌细胞的生存,包括辐照有或没有cucurbitacin b . MCF7:5C mda - mb - 231, SKBR-3细胞治疗<年代vg height="13.7375" id="M16" style="vertical-align:-2.29482pt" version="1.1" viewbox="0 0 33.950001 13.7375" width="33.950001" xmlns="http://www.w3.org/2000/svg">
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</svg>cucurbitacin B辐射前48小时。潜伏期后与特定的药物浓度,细胞被收获,resuspended在新鲜培养基,然后在主/辐照Gy。集落形成检测到21天后,存活曲线。数据是三个实验的总结。</td>
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<h4 id="discussion">4所示。讨论</h4>
<p>Cucurbitacins高含氧,四环三萜烯含有cucurbitane核骨架和极大的兴趣,因为他们表现出的广泛生物活性的植物和动物。已报告Cucurbitacins抑制几种类型的癌症。Cucurbitacins分为十二个类别(<一个href="#B3">20.</一个>]。在各种cucurbitacins,最丰富的是cucurbitacin B (<一个href="#B6">21</一个>]。许多报告表明cucurbitacin B的抗增殖影响乳腺癌细胞。例如,cucurbitacin B,从根中提取和果汁<我>t . cucumerina</我>据报道对人类乳腺癌细胞株的细胞毒性(<一个href="#B16">22</一个>]。Wakimoto et al。<一个href="#B26">9</一个>)报道,cucurbitacin B产生抗癌活动对ER−乳腺癌Her2 / neu放大,p53突变<我>在体外</我>和<我>在活的有机体内</我>(<一个href="#B26">9</一个>]。同样,cucurbitacin B已被报道通过减少Wnt-associated抑制Wnt信号通路蛋白和降低galectin-3-mediated易位<我>β</我>连环蛋白核(<一个href="#B4">23</一个>]。</p>
<p>在这项研究中,我们分析了抗癌活性cucurbitacin B在人类乳腺癌细胞行:MCF7:5C, mda - mb - 231, SKBR-3使用单独生存分析。在这三种细胞行,SKBR-3 Her2 / neu高水平表达的受体,而MCF7:5C和mda - mb - 231并不会表达这些受体。我们发现cucurbitacin B强有力的抗活动在所有类型的细胞。SKBR-3 cucurbitacin B相比是最敏感和其他两个细胞系。我们进一步确定cucurbitacin B细胞周期进程的影响和细胞凋亡诱导乳腺癌细胞系。结果表明,凋亡细胞被cucurbitacin诱导B细胞系,治疗和细胞周期被捕G<年代ub>2</年代ub>/ M阶段MCF7:5C和mda - mb - 231细胞而不是SKBR-3细胞(图<一个href="//www.newsama.com/journals/jo/2012/fig2/" target="_blank">2</一个>)。几个作者表明机体抗癌药物的效果是由于细胞周期改变。microtubule-stabilizing药物紫杉醇,已经被证明可以提高辐射敏感度通过阻断细胞G<年代ub>2</年代ub>/ M期细胞周期(<一个href="#B21">24</一个>,<一个href="#B18">25</一个>]。因为细胞G<年代ub>2</年代ub>已报告/ M期更比其他阶段的细胞周期,对辐射敏感的和cucurbitacin B G治疗增加细胞的数量<年代ub>2</年代ub>/ M期细胞周期,从而增强辐射对乳腺癌细胞株的影响。在我们的研究中,cucurbitacin B施加辐射敏感度当管理5<我>μ</我>辐射对MCF7:5C M cucurbitacin B之前和mda - mb - 231细胞。然而,没有暴露于放射线增减发生当SKBR-3细胞cucurbitacin B在5<我>μ</我>M和辐射,没有积累细胞G<年代ub>2</年代ub>观察/ M期开始之前照射在这些条件下。p21和细胞周期检查站被证明调节核苷酸切除修复过程促进DNA损伤的修复即使没有野生型p53 [<一个href="#B22">26</一个>]。我们的研究结果表明p21表达水平由cucurbitacin B治疗多数显示了所有的细胞系upregulation尤其是SKBR-3显示最高的感应与其他细胞类型(数据相比<一个href="//www.newsama.com/journals/jo/2012/fig4/" target="_blank">4</一个>和<一个href="//www.newsama.com/journals/jo/2012/fig5/" target="_blank">5</一个>)。因此,没有放射线增减清单使用5<我>μ</我>M cucurbitacin B SKBR-3可能因为cucurbitacin B没有引起G<年代ub>2</年代ub>/ M细胞周期阻滞。综上所述,机体效应由cucurbitacin B依赖G的感应<年代ub>2</年代ub>/ M逮捕在乳腺癌细胞,但不一定p21的感应。总之,cucurbitacin B可以增强放射线增减对乳腺癌细胞的影响研究<我>在活的有机体内</我>需要评估cucurbitacin B治疗的生物功效。</p>
<h4 id="conflict-of-interests">利益冲突</h4>
<p>作者报告没有利益冲突。作者仅负责内容和论文的写作。</p>
<h4 id="acknowledgments">确认</h4>
<p>作者要感谢马修·菲茨杰拉德,亚当,约书亚马德森杰出的技术援助。美国Duangmano收到薪水RGJ项目从泰国政府的支持。本文也支持的辐射和自由基核心设施癌症中心提供一部分经费支持格兰特P30 CA086862。</p>
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