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1、Cox-2 inhibition enhances the activity of sunitinib in human renal cell carcinoma xenografts文献赏析文献赏析文献内容框架组成文献内容框架组成2a1I Introduction ntroduction （前言）（前言）2Materials and Materials and Methods Methods （材料与方法）（材料与方法）5OutlookOutlook （展望）（展望）3Results Results （结果）（结果）4Discussion Discussion （讨（讨论）论）Introdu

2、ctionnBackground: Sunitinib (Su), a tyrosine kinase inhibitor of VEGFR, is effective at producing tumour response in clear cell renal cell carcinoma (cRCC), but resistance to therapy is inevitable. As COX-2 is a known mediator of tumour growth, we explored the potential benefit of COX-2 inhibition i

3、n combination with VEGFR inhibition in attempts at delaying tumour progression on Su.nMethods: COX-2 expression was compared with areas of hypoxia in tumours that progressed on Su vs untreated tumours. Mice bearing human cRCC xenografts were treated with Su and the COX-2 inhibitor, celecoxib, and th

4、e effects on tumour growth were assessed. Sequential vs concurrent regimens were compared.nResults: COX-2 expression was increased in cRCC xenografts in areas of tumour hypoxia. The combination of Su and celecoxib achieved longer times to tumour progression compared to treatment with either agent al

5、one or to untreated control animals in four models. This effect was seen with concurrent but not with sequential therapy.nConclusion: COX-2 inhibition can extend the effectiveness of VEGFR inhibition. This effect is dependent on the timing of therapy.nClinical trials combining Su and COX-2 inhibitor

6、s should be considered as a means delaying time to progression on sunitinib in patients with metastatic cRCC.Tumour xenograft inductionA498, 786-O: human cRCC cell lines(ATCC, Manassas, VA, USA), UMRC-3 (Grossman et al, 1985) and MDA-62（a primary tumor)Immunohistochemistry The primary antibodies (an

7、ti-COX-2 antibody anti-Hypoxyprobe antibody ) were applied at a 1 : 50dilution, to sections for 1 h. EnVision System ,HRP labelled polymeranti-mouse ,incubation with DAB chromogen. Tumour perfusion imaging. Tumour perfusion imaging with arterial spin-labelling MRI (ASL MRI) was performed as previous

8、ly described.A 16-colour table was applied in 10ml per 100g per min increments ranging from 0 to 160ml per 100g per min, with flow values represented as varying shades of black, blue, green, yellow, red, and purple, in order of increasing perfusion.Statistical analysisThe primary end point was time

9、to progression (TTP). TTP was defined as time in days to increase by 2mm in tumour size and compared between groups via the exact Wilcoxon rank sum test. Median TTP in days and ranges are reported throughout.Materials and Methods 异种移植异种移植瘤诱导瘤诱导四个实验样本，以雌性裸鼠进行实验A498, 786-O: 人肾癌细胞株(ATCC, Manassas, VA,

10、USA), UMRC-3 (Grossman et al, 1985) and MDA-62（裸鼠体内传代培养三代得到的初级肿瘤)免疫组化免疫组化 一抗(抗Cox-2和抗-Hypoxyprobe )1：50稀释后处理肿瘤切片一小时 ENVISION法免疫组化，HRP标记鼠抗，DAP色素原孵育肿瘤灌注肿瘤灌注成像成像 运用动脉自旋标记（ASL）的核共振成像（MRI）技术的肿瘤灌注成像已经在之前描述过了。用16色的表表示从0到160ml每100g每分钟的范围内10ml每100g每分的增长，同时随着增加灌注而变化的黑色、蓝色、绿色、黄色、红色以及紫色的阴影代表了血流动值。统计学分统计学分析析初始终点

11、是肿瘤进展时间（TTP）。TTP将以肿瘤尺寸增加2mm的时间为准，同时将在用威尔科克森符号秩检验的各组进行对比。天数以及肿瘤尺寸TTP的中值将被始终公布。材材料与方法料与方法Resultsn1、Sunitinib-treated tumours exhibit enhanced hypoxia which correlates with COX-2 expression.n 苏尼替尼处理的肿瘤表现出更强的低氧性，但这个低氧性同时也受COX-2表达的情况影响n2、 Concurrent therapy is superior to sequential therapy.n 同时治疗优于次序性治疗

12、n3、Celecoxib augments the activity of sunitinib in an angiogenesis-independent manner.n 塞来考昔增强了苏尼替尼在抗血管生成方面的活性灌注成像图1、空白组2、塞来昔布3、苏尼替尼4、塞来昔布+苏尼替尼统计学分析统计学分析图三：在额外的人异种移植肾细胞癌模型中，塞来考昔增强了舒尼替尼的抗肿瘤效果。Figure 3. Celecoxib enhances the anti-tumour activity of sunitinib in additional human xenograft clear cell c

13、RCC models. Comparison of volume (mm3) over time (days) following treatment with vehicle, celecoxib, sunitinib, or combination of celecoxib and sunitinib, in two different human xenograft cRCC models: MDA-62 (A) and UMRC-3 (B). N=3 tumours in each arm. Data are shown as averages.e.m图四：转为混合治疗明显比继续舒尼替

14、尼治疗要差。 Figure 4. Continuous sunitinib is superior to sequential therapy. Switching to celecoxib (sunitinib-celecoxib) is significantly worse than continuous sunitinib. Time to increase by an additional 2mm beyond the initial tumour progression on sunitinib is shown: (10 (510) days, n=6 and 12 (1013)

15、 days, n=7, respectively, P=0.003). Delayed addition of celecoxib (sunitinib-celecoxib+sunitinib) (10 (713) days, n=7) did not improve the efficacy of continuous sunitinib (P=0.11). Box plot is read as described in Figure 2C and D.n接下页Figure 5.Measurements of perfusion demonstrate that the celecoxib

16、 does not affect perfusion despite significant tumour size effect. Bar graphs of average tumour perfusion as measured by ASL MRI with SD are shown at four time points in (A). The treatment arms are as follows: sunitinib (n=2), celecoxib (n=2), sunitinibtcelecoxib (n=3), and vehicle (n=3). Representa

17、tive images from the four treatment arms are shown in (B). Representative tumour regions are outlined with yellow line in ASL perfusion images (right column of each time point) and the corresponding reference proton density images (left column). The overall tumour size was measured with long and sho

18、rt axes (in mm), one axis is shown in this figure, and the mean blood flow (in ml per 100 g per min) are shown below each image. A colour bar on bottom represents range of perfusion values from 0 to 160 ml per 100 g per min.n图五：混合疗法对肿瘤血流量有明显的抑制。Discussionn1、Celecoxib as a single agent showed activit

19、y only in the 786-O tumour model.n 单独使用的塞来考昔（celecoxib）只在786-O中有活性n2、However , in all models the combination of sunitinib and celecoxib showed greater inhibition of tumour growth than either agent alone. 苏尼替尼（苏尼替尼（sunitinib）和塞来考昔混用的效果比单用其中任何一种都）和塞来考昔混用的效果比单用其中任何一种都好好n3、The MDA-62 model may be more r

20、elevant to patients as the xenograftMDA-62was obtained directly from a patient undergoing surgery for cRCC, implanted directly into the subcutaneous space of female BALB/c nude mice and serially passaged into new mice without ever being propagated in vitro.n由于直接从患者身上移植到小鼠，未经体外培养，可能具有更高的临床由于直接从患者身上移植

21、到小鼠，未经体外培养，可能具有更高的临床价值价值n4、In general the effects of treatment are more pronounced when tumours are smaller prior to treatment.n肿瘤治疗的优先度越低（越早期），治疗效果越好肿瘤治疗的优先度越低（越早期），治疗效果越好n5、Thus, celecoxib is likely acting by a distinct mechanism from sunitinib,which potently reduces tumour perfusion.n塞塞来稀布可能具有一种来自舒尼替尼的独特机制，可以很大程度上减少来稀布可能具有一种来自舒尼替尼的独特机制，可以很大程度上减少肿瘤肿瘤血流量血流量。Discussionn有待研究之处有待研究之处：n1、由于成像要求不同，对不同的样本进行了不同的预处理，且结果的大致趋势都是一致的。但是，预处理对结果的影响预处理对结果的