potential mechanisms involved in resistant phenotype of MCF-7breast carcinoma cells to ionizing radiation induced apoptosis

In the present study, we investigated the mechanisms of apoptosis resistance and the roles of the phosphorylation of BRCA1, p21, the Bax/Bcl-2 protein ratio and cell cycle arrest in IR-induced apoptosis in MCF-7 cells. X-irradiation, in particular at low dose (1 Gy), but not carbon ion irradiation, had a significant antiproliferative effect on the growth of MCF-7 cells. 1 Gy X-irradiation resulted in G1 and G2 phase arrest, but 4 Gy induced a significant G1 block. In contrast, carbon ion irradiation resulted in a significant accumulation in the G2 phase. Concomitant with the phosphorylation of H2AX induced by DNA damage,carbon ion irradiation resulted in an approximately 1.9–2.8-fold increase in the phosphorylation of BRCA1 on serine residue 1524, significantly greater than that detected for X-irradiation. Carbon ion irradiation caused a dramatic increase in p21 expression and drastic decrease in Bax expression compared with X-irradiation. The data implicated that phosphorylation of BRCA1 on serine residue 1524 might,at least partially, induce p21 expression but repress Bax expression. Together, our results suggested that the phosphorylation of BRCA1 at Ser-1524 might contribute to the G2 phase arrest and might be an upstream signal involved in preventing apoptosis signal via upregulation of p21 and downregulation of the Bax/Bcl-2 ratio.

BRCA1 and its phosphorylation involved in caffeine-inhibitable event upstream of G2 checkpoint

Caffeine, which specifically inhibits ATM/ATR kinases, efficiently abrogates the ionizing radiation (IR)-induced G2 arrest and increases the sensitivity of various tumor cells to IR. Mechanisms for the effect of caffeine remain to be elucidated. As a target of ATM/ATR kinases, BRCA1 becomes activated and phosphorylated in response to IR. Thus, in this work, we investigated the possible role of BRCA1 in the effect of caffeine on G2 checkpoint and observed how BRCA1 phosphorylation was regulated in this process. For these purposes, the BRCA1 protein level and the phosphorylation states were analyzed by Western blotting by using an antibody against BRCA1 and phospho-specific antibodies against Ser-1423 and Ser-1524 residues in cells exposed to a combination of IR and caffeine. The results showed that caffeine down-regulated IR-induced BRCA1 expression and specifically abolished BRCA1 phosphorylation of Ser-1524, which was followed by an override of G2 arrest by caffeine. In addition, the ability of BRCA1 to transactivate p21 may be required for MCF-7 but not necessary for Hela response to caffeine. These data suggest that BRCA1 may be a potential target of caffeine. BRCA1 and its phosphorylation are most likely to be involved in the caffeine-inhibitable event upstream of G2 arrest.

BRCA1对人乳腺癌细胞辐射敏感性的影响及其作用机制研究

目的：研究肿瘤抑制子BRCA1在人乳腺癌细胞辐射抗性中的作用，并探讨其作用机制。材料和方法：利用实时细胞分析系统检测辐射对细胞存活的影响；流式细胞术检测辐射对细胞周期分布的影响；RT-PCR检测辐射导致的BRCA1、Bax和Bcl-2在 mRNA水平变化；Western blot方法检测辐射诱导的蛋白表达水平的变化； In-cell western定量检测辐射引起的蛋白表达水平的变化； AO/EB染色法检测辐射导致的细胞死亡情况。结果：第一，分别用1Gy和4Gy x射线和碳离子束辐照人乳腺癌细胞MCF-7，研究MCF-7对不同LET射线的辐射敏感性差异。结果显示，x射线组1Gy辐射导致细胞存活显著下降，4Gy辐射对细胞存活影响不明显；而碳离子束辐射对细胞生长无抑制作用。与x射线组比较，碳离子辐射诱导了更低的亚“G1”期细胞百分数和更显著的G2期阻滞现象。同时碳离子束辐射诱导BRCA1磷酸化水平和p21表达上调，Bax表达下调。以上结果表明MCF-7对辐射的耐受性与凋亡功能相关，而BRCA1 Ser-1524磷酸化作用可能参与细胞周期和凋亡的信号调控。第二，研究BRCA1在咖啡因诱导的辐射增敏效应中作用。当2mM咖啡因联合4Gy的x射线或碳离子束辐射处理MCF-7细胞后，观察到细胞存活显著下降，辐射诱导的G2期阻滞被废除，BRCA1和p21蛋白表达被抑制，而p53表达水平无明显变化。结果表明咖啡因诱导的MCF-7细胞的辐射增敏作用可能与G2期阻滞被废除相关，BRCA1可能参与该过程的信号调节。第三， 利用BRCA1功能正常的MCF-7细胞和BRCA1功能缺失的HCC1937细胞进一步研究BRCA1对细胞辐射敏感性的影响。辐射显著抑制了HCC1937细胞存活，但对MCF-7细胞存活无明显影响。与HCC1937细胞相比，辐射诱导MCF-7细胞发生显著的G2期阻滞。辐射诱导HCC1937细胞发生晚期凋亡，而MCF-7细胞则多发生早期凋亡，且MCF-7细胞凋亡数明显少于HCC1937细胞。RT-PCR检测结果显示，辐射增强了MCF-7细胞中BRCA1的mRNA 水平，抑制了Bax的mRNA 水平，对Bcl-2的影响不明显；而HCC1937细胞中Bax的mRNA表达水平则被增强。同时辐射诱导MCF-7细胞中BRCA1和p21蛋白表达增强，Bax表达下降，Bcl-2水平略有增高。而HCC1937细胞Bax表达水平增强，但p21和Bcl-2的表达水平则检测不到。这些结果表明，正常的BRCA1功能对Bcl-2的转录表达是必须的，BRCA1通过上调p21水平，下调Bax/Bcl-2影响细胞的辐射敏感性。结论： BRCA1在人乳腺癌细胞的辐射抗性发生中发挥重要作用，BRCA1通过上调p21水平诱导G2期阻滞，下调Bax/Bcl-2抑制凋亡信号，使得细胞对辐射诱导的凋亡产生抗性，最终导致细胞对辐射产生耐受性

黄河三角洲湿地生态系统的形成及其保护

通过对黄河三角洲新生湿地的野外调查,利用TM陆地卫星影像图的判读,结合黄河尾闾变动的历史资料分析,结果表明,黄河三角洲新生湿地是多泥沙的黄河淤积和海岸蚀退所形成.受区域气候、地貌、地质沉积、土壤、植被的相互作用和互相影响,形成4.5×105hm2的湿地,其中有6.84×104hm2的人工湿地.湿地生态类型随着黄河尾闾摆动和海退陆进程度而变化,自海洋向陆地依次为潮下带水生生态系统潮间带湿地生态系统潮上带盐生生态系统芦苇獐茅湿地生态系统草甸湿地生态系统陆上农田生态系统.该系统资源丰富,共有野生动物1524种,鸟类300种,鱼类1040种.石油开发和近年来黄河断流对湿地产生一定影响,应在开发利用中加强管理和保护

沈阳市苏家屯区耕层土壤养分空间变异性研究

利用地统计学和地理信息系统相结合的方法,研究了沈阳市苏家屯区耕地土壤(0～20 cm)有机质、速效氮、速效磷、速效钾含量的空间变异性特征.结果表明,各变量符合正态分布或经对数转换后符合正态分布,最佳的变异函数理论模型均为球状模型.有机质具有较强的空间相关性(C0与C0+C比值为24.11％),速效氮、速效磷、速效钾具有中度的空间相关性(C0与C0+C比值分别为29.53％、60.77％、58.82％).各变量的空间自相关距离分别为26.051、28.293、15.132和23.813 km.有机质和速效氮的空间变异主要受结构性因子(如成土因子)的影响,速效磷和速效钾主要受随机性因子(如施肥)的影响.Kriging插值绘制出的养分含量空问分布图显示了养分的空间分布格局,它将有助于进一步监测养分动态,为农业和环境管理提供数字地图支持.

基于随机背包的公钥密码

该文构造了一个背包型公钥密码算法。该背包公钥密码具有如下优点:加解密只需要加法和模减法运算,因此加解密速度快;该算法是基于随机背包问题而不是易解背包问题而构造的;证明了在攻击者不掌握私钥信息情况下该密码算法能抵抗直接求解背包问题的攻击,包括低密度攻击和联立丢番图逼近攻击等;证明了攻击者能够恢复私钥信息与攻击者能够分解一个大整数是等价的。分析表明,该算法是一个安全高效的公钥加密算法。