NP方案联合不同间隔时间放疗对乳腺癌MCF-7细胞作用研究

目的:研究化疗方案顺铂(DDP) 盖诺(NVB)联合不同间隔时间放射治疗对人乳腺癌MCF-7细胞辐射敏感性影响。方法:采用细胞集落形成方法观察DDP NVB方案联合不同间隔时间放疗对细胞的杀伤作用,流式细胞仪对各组周期分布和凋亡情况进行分析比较,并观察凋亡形态特征。结果:MCF-7细胞化放疗间隔不同时间后细胞SF值差异随照射剂量增加而逐渐明显,8Gy时12小时组最低,0小时组居中,48、72小时组最高。化放疗间隔时间不同引起MCF-7细胞SF值不同与NP作用后不同间隔时间G2/M期阻滞比例不同有一定相关性,NP化疗可诱导细胞凋亡,而且随时间增加呈不可逆转增加,但化疗后不同时间凋亡差异同SF值差异间并未呈现相关性。结论:NP不同时机顺序联合放疗对MCF-7细胞辐射敏感性有一定影响,这种差异同NP作用后不同时间细胞周期分布、凋亡等作用有关。     

盖诺与顺铂联合化疗加同期放射对乳腺癌MCF-7细胞作用的研究

目的：探讨盖诺（NVB）联合顺铂（DDP）的放化同期治疗乳腺癌的有效性、时机选择和作用机制。方法：NVB和DDP（NP方案）对乳腺癌MCF-7细胞联合化疗同期放射。比较照射加联合化疗组和单纯照射组、不同间隔时间联合化放疗的细胞存活率（SF）、凋亡和细胞周期阻滞差异。结果：联合化疗加同期放射后SF较单独放疗更低。化疗处理后间隔4、12和36h照射的SF值最低,0h居中,而阃隔48和72h后,SF值明显上升,放化疗协同杀灭效应减弱,SF最低与最高值相差约15倍。联合化疗加照射组细胞凋亡指数（AI）明显高于单独放射和联舍化疗组。6、8Gy放射后36h时,放化疗联合组与单独放射组比较A1分别高达30．7％、38．35％和2．22％、3．27％,差异有统计学意义,P〈0．05。化疗处理MCF-7细胞1h后在间隔4、12、36和72h处,G2／M周期细胞比例较高,0.48h处较低。结论：联合化放疗对乳腺癌MCF-7细胞具有较强的协同杀灭作用,化放联合治疗的时机选择在肿瘤细胞杀灭过程中具有重要的意义,这种协同杀灭作用与增加细胞凋亡率和化疗造成的细胞G2／M周期阻滞有关。     

红参含药血清对人乳腺癌MCF?鄄7细胞增殖和细胞周期的影响

[目的]研究红参含药血清对人乳腺癌MCF-7细胞增殖和细胞周期的影响.[方法]采用MTT法观察红参含药血清对人乳腺癌细胞MCF-7的增殖活性.流式细胞术检测人乳腺癌细胞MCF-7的细胞周期、细胞凋亡率.[结果]红参低剂量含药血清对MCF-7细胞有促增殖作用,24、48、72h时增殖率分别为105.9％、115.9％和121.5％.而高剂量对MCF-7细胞表现为抑制作用.红参低剂量组使MCF-7细胞S期DNA含量比例明显增多,高剂量组使细胞周期阻滞在G0/G1期,并能够诱导细胞凋亡,凋亡率达26.86％.[结论]红参含药血清在高剂量时抑制MCF-7细胞生长,低剂量时对细胞具有促增殖作用.     

NP方案联合不同间隔时间放疗对乳腺癌MCF-7细胞凋亡影响的研究

目的：研究NP方案[顺铂（DDP）＋盖诺（NVB）]联合不同间隔时间放射治疗对乳腺癌MCF-7细胞凋亡的影响。方法：采用流式细胞仪技术定性定量研究NP方案联合不同间隔时间放疗对人乳腺癌MCF-7细胞凋亡的影响，并观察NP方案作用后不同时间周期阻滞情况。结果：MCF-7细胞单独NP方案化疗组或放化联合作用组均可诱导细胞凋亡，随时间增加呈不可逆转增加，放化疗组凋亡指数明显高于单独化疗组，P〈0．05。NP化疗后不同间隔时间G2／M期阻滞比例不同。结论：化放不同间隔组凋亡指数随间隔时间延长而增加，间隔72h达到峰值。     

（125）I联合ADM对乳腺癌MCF-7细胞增殖、凋亡的影响

目的研究放射性粒子125I联合化疗药物多柔比星（阿霉素,ADM）对乳腺癌MCF-7细胞增殖、凋亡的影响。方法按2×2析因设计将乳腺癌敏感株MCF-7细胞随机分成A、B、C、D 4组,A组：空白对照组;B组：单纯125I粒子组;C组：单纯ADM组;D组：125I粒子＋ADM组。采用流式细胞术检测各组干预后的细胞周期分布及细胞凋亡率。结果 （1）单纯125I粒子近距离低剂量率持续照射后将MCF-7细胞阻滞于G2~M期1。25I联合ADM将MCF-7细胞周期主要集中在G0~G1期,同时伴有较大比例的细胞凋亡。（2）各组细胞的早期凋亡率分别为：A组（0.99±0.05）%、B组（19.22±4.92）%、C组（16.57±4.73）%、D组（3.16±1.08）%;各组晚期凋亡及坏死率为：A组（0.32±0.18）%、B组（3.16±1.39）%、C组（3.24±0.75）%、D组（28.99±7.96）%,D组晚期凋亡及死亡率明显提高。结论 125I放射性粒子能有效诱导乳腺癌细胞凋亡,与化疗药物ADM联合作用除诱导细胞凋亡,还可导致大量细胞死亡,具有协同、增效的作用。     

阿霉素对人乳腺癌细胞凋亡和增殖的影响

[目的]检测阿霉素(ADR)对体外人乳腺癌化疗敏感细胞(MCF-7/S)凋亡和增殖的影响,并探讨其可能的作用机理.[方法]应用MTT比色法检测ADR对体外培养的MCF-7/S细胞增殖抑制作用,应用流式细胞术检测ADR诱导乳腺癌细胞凋亡,采用免疫细胞化学法检测ADR作用前后去磷酸化Rb蛋白和增殖细胞核抗原(PCNA)的表达.[结果]ADR抑制MCF-7/S细胞增殖,呈剂量依赖性;ADR组MCF-7/S细胞的凋亡率、磷酸化Rb蛋白的表达量与对照组相比明显增高(P＜0.01);PCNA阳性表达率与对照组的相比明显降低(P＜0.01).在ADR组,MCF-7/S细胞的凋亡率(AR)与去磷酸化Rb蛋白的表达量呈正相关,与增殖细胞核抗原的阳性表达率呈负相关.[结论]ADR诱导MCF-7/S细胞凋亡并抑制MCF-7/S细胞增殖可能与其上调去磷酸化Rb蛋白和下调细胞内增殖细胞核抗原表达水平有关.     

X线照射后对乳腺癌细胞凋亡的影响及CDKN1A表达的变化

目的探讨乳腺癌细胞系（MCF-7）X线照射后CDKN1A基因（p21）表达变化及其对细胞凋亡的影响。方法用流式细胞方法检测MCF-7细胞在接受不同剂量X射线照射后CDKN1A基因表达的改变和用RNAi技术抑制CDKN1A基因表达并检测细胞凋亡的变化。结果MCF-7细胞在接受不同剂量（1、2和4 Gy） X射线照射后CDKN1A蛋白表达有不同程度升高,其中 4 Gy照射后升高水平最明显（P<0.05）。在接受4 Gy剂量照射后,CDKN1A蛋白水平在8、12、24、48、72 h 均有不同程度增加,其中24 h时较对照组升高3倍(P<0.05)。抑制CDKN1A基因表达后MCF-7细胞凋亡率增加183.9%（P<0.05）。结论乳腺癌细胞在接受4 Gy照射后24 h的CDKN1A表达水平增加最为明显,抑制CDKN1A基因表达可促进细胞凋亡。     

别直参抑制人乳腺癌细胞MCF-7体外增殖

[目的]评价别直参对MCF-7细胞体外增殖的影响,并检测其对凋亡相关蛋白bax/bcl-2表达的影响.[方法]采用MTT比色法检测对MCF-7细胞增殖的作用;流式细胞术检测对MCF-7细胞凋亡的影响;免疫组化法检测bax/bcl-2的表达.[结果]别直参高剂量组作用MCF-7细胞24h后,MTT检测增殖率为89.83％,提示有抑制MCF-7细胞体外增殖的作用(P＜0.05);流式细胞术检测高剂量组细胞凋亡率明显升高(P＜0.05);免疫组化法检测高剂量组能上调MCF-7细胞bax的表达,并下调bcl-2的表达.[结论]别直参高剂量组具有一定的诱导MCF-7细胞凋亡效应作用,其机制可能通过上调bax的表达和下调bcl-2的表达.     

全脑放疗联合伽玛刀治疗脑转移瘤

[目的]评价全脑放疗联合伽玛刀治疗脑转移瘤的价值.[方法]将60例脑转移瘤病人随机分为两组:全脑放疗联合伽玛刀治疗组(A组)及单纯伽玛刀治疗组(B组).A组先全脑放疗40Gy/20F/4W后,复查颅脑MRI或CT,如果有残留,则继以伽玛刀治疗,周边剂量8Gy～26Gy,一次照射:B组单纯伽玛刀治疗,肿瘤周边剂量16Gy～32Gy,一次照射.分别对两组不同病灶数患者的近期疗效及生存率进行统计学分析.[结果]两组患者近期疗效及生存率差异无显著性;分层研究显示:单个病灶者两组生存率差异无显著性,而两个以上病灶者全脑放疗联合伽玛刀治疗的生存率高于单纯伽玛刀治疗,差异有显著性.[结论]对于单个病灶的脑转移瘤可单纯行伽玛刀治疗:而多发病灶的脑转移瘤,建议全脑放疗联合伽玛刀治疗.     

Apatinib联合5-Fu协同抑制乳腺癌MCF-7细胞的体外研究

目的 探讨阿帕替尼(Apatinib)联合5-氟尿嘧啶(5-Fu)对乳腺癌MCF-7细胞的抑制作用.方法 采用人乳腺癌细胞株MCF-7,首先采用MTr法及应用流式细胞仪测定不同浓度apatinib作用后对其细胞增殖和周期的影响,其次将MCF-7细胞分为4组,即对照组、Apatinib单药组、5-Fu单药组、Apanitib+ 5-Fu联合组.对各组细胞给予相应药物处理,48 h后分别应用流式细胞仪检测各组药物对MCF-7细胞株凋亡的作用.结果 Apatinib单药对MCF-7细胞有增殖抑制作用,且存在时间剂量依赖关系,而对其细胞周期影响不大.与对照组相比,Apatinib联合5-Fu作用后有协同诱导凋亡作用,经流式细胞仪检测,Apatinib组诱导的细胞凋亡率为12.05％,5-Fu组为25.76％.与单药组比,Apatinib+5-Fu联合组凋亡率升高更为明显,达34.90％ (P＜ 0.05).结论 Apatinib和5-Fu的联合应用在体外协同抑制乳腺癌MCF-7细胞并诱导凋亡,使抗肿瘤活性显著增强.     

Prognostic Significance of Inflammation-associated Blood Cell Markers in Nonmetastatic Clear Cell Renal Cell Carcinoma

ObjectivesOur objective was to evaluate the effect of the neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), lymphocyte/monocyte ratio (LMR), and red blood cell distribution width (RDW) on the survival outcomes of nonmetastatic clear cell renal cell carcinoma (ccRCC).Materials and MethodsWe accessed our single-center, urologic-oncologic registry to extract the data for patients who had undergone nephrectomy for nonmetastatic ccRCC. The optimal cutoff for these markers was determined using X-tile software, and survival analyses using Cox regression were performed.ResultsA total of 687 patients had undergone nephrectomy. The optimal cutoffs for NLR, PLR, LMR, and RDW were 3.3, 210, 2.4, and 14.3%, respectively. The NLR, PLR, LMR, and RDW were significantly associated with a larger pathologic tumor size, and stage, more aggressive Fuhrman grade, and the presence of tumor necrosis. After adjusting for age, baseline Eastern Cooperative Oncology Group, pathologic tumor and nodal stage, and Fuhrman grade, only PLR remained an independent prognostic marker for both cancer-specific survival (hazard ratio, 2.69; 95% confidence interval, 1.36-5.33; P = .004) and overall survival (hazard ratio, 2.19; 95% confidence interval, 1.36-3.50; P = .001). When the PLR was included with the Leibovich score and University of California, Los Angeles, integrated staging system, the Harrell’s c-index increased from 0.854 to 0.876 and 0.751 to 0.810, respectively, for cancer-specific survival at 5 years after nephrectomy. When risk stratified by the Leibovich risk group and UCLA integrated staging system, PLR was a significant prognostic factor only within the intermediate- to high-risk groups.ConclusionsPLR is a robust prognostic marker in nonmetastatic ccRCC that clearly outperforms other inflammatory indexes in those who had undergone nephrectomy. However, its prognostic effect was limited in the low-risk category of ccRCC.     

Cell kinetics

Cell kinetic concepts have pervaded radiation therapy since the early part of the 20th century and have been instrumental in the development of modern radiotherapy. In this review, the fundamental radiobiological concepts that have been developed based on cell kinetic knowledge will be revisited and discussed in the context of contemporary radiation therapy. This will include how the proliferation characteristics, variation in sensitivity during the cell cycle and the extent of radiation-induced cell cycle delay translate into a variable time for the expression of damage, how cell kinetics interacts with hypoxia and how the response to fractionated radiation schedules is influenced by cell kinetics in terms of repair, redistribution, reoxygenation and repopulation. The promise of combining radiation with new biologically targeted agents and the potential of non-invasive positron emission tomography imaging of proliferation are areas where cell kinetics will continue to influence radiotherapy practice.     

Cell Size Following Irradiation in Relation to Cell Cycle

The cell volume of Ehrlich ascites tumour cells was studied following a radiation dose of 5.0 Gy. The cell volume increased 12 to 30 hours after irradiation by about 20 per cent, was normal at about 50 hours, and increased again at 72 hours. In order to explain these changes the composition of the cells in cell cycle was studied. In addition, the cell volume of irradiated cells from the various parts of the cell cycle, separated by centrifugal elutriation, was measured. The changes in the mean cell volume of unseparated cells could be explained by variations in the cell cycle composition of the cell population. Irradiated cells from the various parts of the cell cycle did not deviate from the volume of non-irradiated cells. The cell volume doubled during the cell cycle. This increase was, however, not linear.     

Inhibition of Glioma Cell Proliferation by Neural Stem Cell Factor

Summary Neural stem cells (NSC) have unique differentiation-, proliferation-, and motility properties. To investigate whether they secrete factors that interfere with the proliferation of glioma cells, we grew glioma cells in conditioned medium (CM) obtained from cultures of neurospheres including neural stem / progenitor cells (NSPC) isolated from embryonic (E14)- or adult mouse brain or fetal human brain. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and BrdU-labeling assays showed that CM from NSPC (NSPC/CM) contained factor(s) that inhibited the proliferation of glioma cells by 28–87%. Filter-fractionation of NSPC/CM revealed that the 50,000–100,000 nominal molecular weight limit (NMWL) fraction contained the inhibitory activity. On the basis of these observations we transplanted 203G glioma cells and/or NSPC into the intrathecal space of the cisterna magna of mice to investigate whether NSPC interfere with the proliferation of glioma cells in vivo. Mice transplanted with both 203G and NSPC survived significantly longer than did mice transplanted only with 203G. We concluded that NSPC secrete factor(s) that may control glioma cell proliferation.     

High Occurrence of Non-Clear Cell Renal Cell Carcinoma in Oman

It is conventionally accepted that renal cell carcinoma (RCC) occurs in older patients and the clear cell type is the most common histology. However, ethnic variations exist and this study was carried out to determine the epidemiological pattern of RCC in Oman. Ninety RCC patients who presented to a tertiary care center in the Sultanate of Oman from 2010 to 2014 were studied. The main findings were that the median age of presentation was low, more patients presented with localized stage, and there was a higher incidence of non-clear (especially papillary) histology. Data from other Gulf countries and possible reasons for the different profile are discussed.     

Changes in Cell Characteristics Due to Culture Conditions in Cell Lines From Human Small Cell Lung Cancer

Eight cultured cell lines were established from human smallcell lung cancers. Every cell line showed the morphologicaland biochemical characteristics of small cell cancer. Changesin cell characteristics were observed in many of these celllines when culture conditions were changed: "oat cell type"changed to "intermediate cell type" and vice versa when serum-freemedium was changed to serum-supplemented medium; a deficiencyof vitamin A in the medium caused a change to squamous cellsand vice versa; and a tumor promoter (teleocidin B) enhancedthe adherence of these cells to the surface of plastic culturedishes. These findings provide evidence that many small celllung cancer cell lines can change their morphology with changesin the environment of the cells.     

Inhibition of Natural Killer Cell Cytotoxicity by Cell Growth-related Molecules

Certain MHC class I molecules on target cells are known to inhibit the cytotoxic action of NK cells. By using monoclonal antibody (mAb) Cho-1, we have found inhibitory non-MHC class I cell surface molecules that are noncovalently-associated with 200 kDa and 40 kDa antigens. Poly I-C-induced rat NK cells were not cytotoxic to rat fetus-derived fibroblast WFB cell line. In contrast, NK cells were cytotoxic to H- ras oncogene-induced transformants of WFB, W14 and W31. FACS analysis indicated that mAb Cho-1 reacts with WFB, but not with W14 and W31 cells. Thus, this antigen may disappear concomitantly with cell growth and transformation. Cho-1 antigens were also expressed on other NK-resistant lines, such as mouse BALB3T3 fibroblast, EL-4 lymphoma and human fibroblast HEPM. However, they were not expressed on NK-sensitive mouse YAC-1 and H- ras transformant (Brash) of BALB3T3 cells. Furthermore, treatment of target cells with IFN-γ clearly induced the cell surface expression of Cho-1 antigens, and conferred a resistance to NK cytolysis on target cells. These data strongly suggest that Cho-I antigen expression may correlate with target cell susceptibility to NK cells. Indeed, treatment of NK-resistant WFB as well as HEPM cells with F(ab')₂ fragments of mAb Cho-1 resulted in the acquisition of susceptibility to NK cytolysis. Cho-1 antigens may be novel molecules that regulate the NK resistance of cells.