中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2010年
8期
580-585
,共6页
袁静%吴春娃%刘竹君%魏熙胤%李凯
袁靜%吳春娃%劉竹君%魏熙胤%李凱
원정%오춘왜%류죽군%위희윤%리개
恩度%多西紫杉醇%给药顺序%循环内皮细胞%肺腺癌细胞系A549%裸鼠
恩度%多西紫杉醇%給藥順序%循環內皮細胞%肺腺癌細胞繫A549%裸鼠
은도%다서자삼순%급약순서%순배내피세포%폐선암세포계A549%라서
Endostar%Docetaxel%Administration sequence%Circulating endothelial cells (CECs)%Lung cancer cell line,A549%Nude mice
目的 观察重组人血管内皮抑制素注射液(商品名:恩度)与多西紫杉醇联合使用时不同给药顺序的抗肿瘤效应.方法 建立肺腺癌A549荷瘤裸鼠模型,随机分为3组,每组8只.(1)同时用药组:每只小鼠恩度400μg/d,第1～35天;多西紫杉醇10 mg/kg,第1～19天,每3 d给药1次.(2)先恩度组:每只小鼠恩度400 μg/d,第1～35天;多西紫杉醇10 mg/kg,第16～34天,每3 d给药1次.(3)模型组:每只小鼠生理盐水100 μl/d,第1～35天;注射用水200 μl/d,第1～35天,每3 d注射1次.同时设立空白对照组(未荷瘤的正常裸鼠,8只),注射方法同模型组.实验过程中测量各组裸鼠体重、移植瘤体积,并计算抑瘤率.实验结束后,采用流式细胞术检测裸鼠外周血循环内皮细胞(CECs)数,采用免疫组化法检测移植瘤组织中基质金属蛋白酶2(MMP-2)、MMP-9、MMP-2的抑制剂(TIMP-2)、MMP-9的抑制剂(TIMP-1)、细胞外MMP诱导因子(EMMPRIN)、α-平滑肌肌动蛋白(SMA)的表达情况,并计数微血管密度(MVD).结果 同时用药组肿瘤体积增长为39.94 mm3,先恩度组肿瘤体积增长为(99.57±74.48)mm3,二者均明显小于模型组[(217.67±95.44)mm3,均P＜0.05].同时用药组、先恩度组、模型组和空白对照组裸小鼠外周血中CECs的数量分别为(25.86±11.77)个/104个细胞、(77.25±24.02)个/104个细胞、(14.71±11.07)个/104个细胞和(12.90±11.20)个/104个细胞,同时用药组、模型组和空白对照组均明显低于先恩度组(均P＜0.01).同时用药组和先恩度组移植瘤组织中MMP-2和MMP-9的表达均较模型组下调(均P＜0.05),先恩度组移植瘤组织中TIMP-1和同时用药组TIMP-2的表达均较模型组上调(均P＜0.05),同时用药组EMMPRIN的表达较模型组下调(P＜0.05).同时用药组和先恩度组移植瘤组织的MVD及α-SMA的水平均低于模型组(均P＜0.05).结论 同时用药组的抑瘤效果及小鼠生存质量更好.同时用药组和先恩度组均可通过下调MMPs的表达以限制肿瘤进展.CECs的变化可能是先上升后下降的动态过程,早期升高可能提示新生血管床面积减小,继而下降则反映联合治疗后的CECs凋亡和肿瘤消退,动态观察CECs的变化可能有助于了解肿瘤负荷及其血管床的变化并预测疗效.
目的 觀察重組人血管內皮抑製素註射液(商品名:恩度)與多西紫杉醇聯閤使用時不同給藥順序的抗腫瘤效應.方法 建立肺腺癌A549荷瘤裸鼠模型,隨機分為3組,每組8隻.(1)同時用藥組:每隻小鼠恩度400μg/d,第1～35天;多西紫杉醇10 mg/kg,第1～19天,每3 d給藥1次.(2)先恩度組:每隻小鼠恩度400 μg/d,第1～35天;多西紫杉醇10 mg/kg,第16～34天,每3 d給藥1次.(3)模型組:每隻小鼠生理鹽水100 μl/d,第1～35天;註射用水200 μl/d,第1～35天,每3 d註射1次.同時設立空白對照組(未荷瘤的正常裸鼠,8隻),註射方法同模型組.實驗過程中測量各組裸鼠體重、移植瘤體積,併計算抑瘤率.實驗結束後,採用流式細胞術檢測裸鼠外週血循環內皮細胞(CECs)數,採用免疫組化法檢測移植瘤組織中基質金屬蛋白酶2(MMP-2)、MMP-9、MMP-2的抑製劑(TIMP-2)、MMP-9的抑製劑(TIMP-1)、細胞外MMP誘導因子(EMMPRIN)、α-平滑肌肌動蛋白(SMA)的錶達情況,併計數微血管密度(MVD).結果 同時用藥組腫瘤體積增長為39.94 mm3,先恩度組腫瘤體積增長為(99.57±74.48)mm3,二者均明顯小于模型組[(217.67±95.44)mm3,均P＜0.05].同時用藥組、先恩度組、模型組和空白對照組裸小鼠外週血中CECs的數量分彆為(25.86±11.77)箇/104箇細胞、(77.25±24.02)箇/104箇細胞、(14.71±11.07)箇/104箇細胞和(12.90±11.20)箇/104箇細胞,同時用藥組、模型組和空白對照組均明顯低于先恩度組(均P＜0.01).同時用藥組和先恩度組移植瘤組織中MMP-2和MMP-9的錶達均較模型組下調(均P＜0.05),先恩度組移植瘤組織中TIMP-1和同時用藥組TIMP-2的錶達均較模型組上調(均P＜0.05),同時用藥組EMMPRIN的錶達較模型組下調(P＜0.05).同時用藥組和先恩度組移植瘤組織的MVD及α-SMA的水平均低于模型組(均P＜0.05).結論 同時用藥組的抑瘤效果及小鼠生存質量更好.同時用藥組和先恩度組均可通過下調MMPs的錶達以限製腫瘤進展.CECs的變化可能是先上升後下降的動態過程,早期升高可能提示新生血管床麵積減小,繼而下降則反映聯閤治療後的CECs凋亡和腫瘤消退,動態觀察CECs的變化可能有助于瞭解腫瘤負荷及其血管床的變化併預測療效.
목적 관찰중조인혈관내피억제소주사액(상품명:은도)여다서자삼순연합사용시불동급약순서적항종류효응.방법 건립폐선암A549하류라서모형,수궤분위3조,매조8지.(1)동시용약조:매지소서은도400μg/d,제1～35천;다서자삼순10 mg/kg,제1～19천,매3 d급약1차.(2)선은도조:매지소서은도400 μg/d,제1～35천;다서자삼순10 mg/kg,제16～34천,매3 d급약1차.(3)모형조:매지소서생리염수100 μl/d,제1～35천;주사용수200 μl/d,제1～35천,매3 d주사1차.동시설립공백대조조(미하류적정상라서,8지),주사방법동모형조.실험과정중측량각조라서체중、이식류체적,병계산억류솔.실험결속후,채용류식세포술검측라서외주혈순배내피세포(CECs)수,채용면역조화법검측이식류조직중기질금속단백매2(MMP-2)、MMP-9、MMP-2적억제제(TIMP-2)、MMP-9적억제제(TIMP-1)、세포외MMP유도인자(EMMPRIN)、α-평활기기동단백(SMA)적표체정황,병계수미혈관밀도(MVD).결과 동시용약조종류체적증장위39.94 mm3,선은도조종류체적증장위(99.57±74.48)mm3,이자균명현소우모형조[(217.67±95.44)mm3,균P＜0.05].동시용약조、선은도조、모형조화공백대조조라소서외주혈중CECs적수량분별위(25.86±11.77)개/104개세포、(77.25±24.02)개/104개세포、(14.71±11.07)개/104개세포화(12.90±11.20)개/104개세포,동시용약조、모형조화공백대조조균명현저우선은도조(균P＜0.01).동시용약조화선은도조이식류조직중MMP-2화MMP-9적표체균교모형조하조(균P＜0.05),선은도조이식류조직중TIMP-1화동시용약조TIMP-2적표체균교모형조상조(균P＜0.05),동시용약조EMMPRIN적표체교모형조하조(P＜0.05).동시용약조화선은도조이식류조직적MVD급α-SMA적수평균저우모형조(균P＜0.05).결론 동시용약조적억류효과급소서생존질량경호.동시용약조화선은도조균가통과하조MMPs적표체이한제종류진전.CECs적변화가능시선상승후하강적동태과정,조기승고가능제시신생혈관상면적감소,계이하강칙반영연합치료후적CECs조망화종류소퇴,동태관찰CECs적변화가능유조우료해종류부하급기혈관상적변화병예측료효.
Objective To observe and analyze the antitumor effect of endostar combined with docetaxel under different administration sequences. Methods Nude mice with xenograft tumor( A549 cell line) were randomized into 3 groups, 8 mice/group: ( 1 ) Concurrent administration group ( each mouse:endostar400 μg/d, d1-d35, docetaxel 10 mg/kg, every 3 days, d1-d19); (2)Endo-first group (each mouse: endostar 400 μg/d, d1-d35, docetaxel 10 mg/kg, every 3 days, d16-d34); (3)Model group (positive control, tumor-bearing mice without treatment, each mouse: physiological saline, 100 μl/d, d1-d35, water for injection, 200 μl/d, d1-d35, every 3 days), and blank control group ( negative control,normal mice without treatment, 8 mice), the administration method was the same to the model group. The volume of tumor and the weight of mouse were measured during treatment. Circulating endothelial cells (CECs) were detected by flowcytometry, and the expression of matrix metalloproteinase (MMP-2, MMP-9), the tissue inhibitor of MMP (TIMP-1, TIMP-2), the extracellular MMP inducer (EMMPRIN), CD34,α-smooth muscle actin (α-SMA) were determined by immunohistochemistry. Results The tumor growth of concurrent administration group( 39.94 mm3) was lower than that of the endo-first group[(99.57 ± 74.48 )mm3]during treatment, both of them were smaller than that of the model group[(217.67±95.44) mm3,P ＜0.05]. The amount of CECs in the endo-first group[( 77.25 ± 24.02 ) cells/104cells]was more than that of the concurrent administration group[(25.86 ± 11.77) cells/104 cells], the model group[( 14.71 ±11.07 ) cells/104 cells], and the blank control group[( 12.90 ± 11.20 ) cells/104 cells, P ＜ 0.01]. The expression of MMPs in the treatment groups was obviously downregulated. The expressions of TIMP-1 in the endo-first group and TIMP-2 in the concurrent administration group were upregulated ( P ＜ 0. 05 ). The expression of EMMPRIN was significantly down-regulated in the concurrent administration group (P ＜0.05). The MVD and α-SMA expressions of the treatment groups were less than that of the model group (P ＜0.05). Conclusion In comparison with the endo-first group, the anti-tumor effect and survival quality of the concurrent administration group are better. Both of the administration groups may have "vascular normalization effect" by down-regulating MMPs expression through different points, and inhibit the cancer-induced stromal reaction, restraining the cancer progress to a certain extent. The changes of CECs should be a dynamic process with an initial rise in the early-stage suggesting the decrease of vascular bed and subsequent decline ascribed to apoptosis of CECs and the tumor-regression after combined therapy.Investigation of its dynamic changes may be helpful to know the change of tumor burden and vascular bed and predict the antitumor effect.
