目的:观察参芪扶正注射液通过免疫调节提高顺铂化疗的敏感性,并探讨其作用机制。方法:采用噻唑蓝(MTT)比色法测定参芪扶正注射液1,10,100 m L·L^-1对细胞的敏感性,流式细胞法检测细胞凋亡率,酶联免疫印迹法(Western blot)检测凋亡...目的:观察参芪扶正注射液通过免疫调节提高顺铂化疗的敏感性,并探讨其作用机制。方法:采用噻唑蓝(MTT)比色法测定参芪扶正注射液1,10,100 m L·L^-1对细胞的敏感性,流式细胞法检测细胞凋亡率,酶联免疫印迹法(Western blot)检测凋亡相关蛋白的表达,酶联免疫吸附实验(ELISA)检测细胞因子表达水平。结果:参芪扶正注射液能显著抑制人乳腺癌细胞MDA-MB-231与THP-1共培养细胞生长,参芪扶正注射液10,100 m L·L^-1时,其细胞存活率分别为71.8%,59.9%;参芪扶正注射液10 m L·L^-1与顺铂联合运用,提高了共培养细胞对顺铂的敏感性,顺铂的半数抑制浓度(IC50)由30μmol·L^-1降低至15μmol·L^-1;联合运用参芪扶正注射液10 m L·L^-1和顺铂,比15μmol·L^-1顺铂诱导的细胞凋亡率增加了15.5%(P<0.05);联合用药均能明显抑制B细胞淋巴瘤-2(Bcl-2),B细胞淋巴瘤-w(Bcl-w)和细胞外大淋巴瘤(Bcl-xl)的表达,增加Bcl-2相关X蛋白(Bax)和人BH3结构域凋亡诱导蛋白(Bid)的表达(P<0.05);参芪扶正注射液降低顺铂诱导的白细胞介素-10(IL^-10)和前列腺素E2(PGE2)的释放(P<0.05)。结论:参芪扶正注射液通过调节免疫细胞改善了乳腺癌细胞MDA-MB-231对顺铂的敏感性,起到协同抑制肿瘤细胞增殖的作用。该研究为益气扶正防治肿瘤提供实验依据,为中医药缓解肿瘤耐药研究提供实验参考。展开更多
AIM:To investigate the proteolytic contribution of tumor-associated macrophages(TAM)in tumor invasion,we analyzed whether TAM at the invasive front of small HCC in Abcb4-/--mice show an enhanced expression of MMP-9. M...AIM:To investigate the proteolytic contribution of tumor-associated macrophages(TAM)in tumor invasion,we analyzed whether TAM at the invasive front of small HCC in Abcb4-/--mice show an enhanced expression of MMP-9. METHODS:Liver cryosections of the hepatocellular carcinoma(HCC)invasive front from 12 mo old Abcb4-/--mice were stained for collagen typeⅠand MMP-9 using Alexa488 and Alexa568 labeled secondary antibodies.Afterwards,the Alexa568 dye was bleached and the macrophage marker F4/80 was visualized using Alexa568 labeled secondary antibodies.Finally, photographs of the invasive tumor front were digitally overlaid and analyzed. RESULTS:After complete bleaching of the primary dye,specific fluorescence staining of a third antigen, here F4/80,was successfully performed on the same histological section.With this method,we were able to identify conglomerates of matrix metalloproteinase (MMP-9)expressing macrophages within the tumor capsule of HCC. CONCLUSION:MMP-9 expressing macrophages are involved in matrix remodelling at the invasive tumor front of HCC.The described staining protocol provides a simple yet powerful extension of conventional immuno-histochemistry,facilitating visualization of at least three different antigens plus nuclei in one single histological section.展开更多
目的研究胃癌中是否存在肿瘤微环境转移(tumor microenvironment of metastasis,TMEM)及其与胃癌血道转移的相关性。方法采用免疫组化双染法同时标记巨噬细胞和内皮细胞,计数26例远处转移及其配对26例无远处转移胃癌组织中TMEM密度。结...目的研究胃癌中是否存在肿瘤微环境转移(tumor microenvironment of metastasis,TMEM)及其与胃癌血道转移的相关性。方法采用免疫组化双染法同时标记巨噬细胞和内皮细胞,计数26例远处转移及其配对26例无远处转移胃癌组织中TMEM密度。结果 (1)TMEM存在于胃癌组织中;(2)TMEM与分化程度、临床分期明显相关(P<0.001,P=0.006),与肿瘤大小、Lauren分型、淋巴结转移等其他临床病理特点无关(P均>0.05);(3)TMEM与有无远处转移呈正相关(P<0.001)。结论证实胃癌中存在TMEM,TMEM密度可以预测胃癌血道转移,有望成为评估胃癌血道转移危险性的新预测指标。展开更多
文摘目的:观察参芪扶正注射液通过免疫调节提高顺铂化疗的敏感性,并探讨其作用机制。方法:采用噻唑蓝(MTT)比色法测定参芪扶正注射液1,10,100 m L·L^-1对细胞的敏感性,流式细胞法检测细胞凋亡率,酶联免疫印迹法(Western blot)检测凋亡相关蛋白的表达,酶联免疫吸附实验(ELISA)检测细胞因子表达水平。结果:参芪扶正注射液能显著抑制人乳腺癌细胞MDA-MB-231与THP-1共培养细胞生长,参芪扶正注射液10,100 m L·L^-1时,其细胞存活率分别为71.8%,59.9%;参芪扶正注射液10 m L·L^-1与顺铂联合运用,提高了共培养细胞对顺铂的敏感性,顺铂的半数抑制浓度(IC50)由30μmol·L^-1降低至15μmol·L^-1;联合运用参芪扶正注射液10 m L·L^-1和顺铂,比15μmol·L^-1顺铂诱导的细胞凋亡率增加了15.5%(P<0.05);联合用药均能明显抑制B细胞淋巴瘤-2(Bcl-2),B细胞淋巴瘤-w(Bcl-w)和细胞外大淋巴瘤(Bcl-xl)的表达,增加Bcl-2相关X蛋白(Bax)和人BH3结构域凋亡诱导蛋白(Bid)的表达(P<0.05);参芪扶正注射液降低顺铂诱导的白细胞介素-10(IL^-10)和前列腺素E2(PGE2)的释放(P<0.05)。结论:参芪扶正注射液通过调节免疫细胞改善了乳腺癌细胞MDA-MB-231对顺铂的敏感性,起到协同抑制肿瘤细胞增殖的作用。该研究为益气扶正防治肿瘤提供实验依据,为中医药缓解肿瘤耐药研究提供实验参考。
基金Supported by the Grants from the Deutsche Forschungs-gemeinschaft(RO 957/8-1 and SFB/TRR 57)and by BMBF ZooMAP-TPC4a Research Grant of the University Medical Center Giessen and Marburg(UKGM 10/2010 GI)
文摘AIM:To investigate the proteolytic contribution of tumor-associated macrophages(TAM)in tumor invasion,we analyzed whether TAM at the invasive front of small HCC in Abcb4-/--mice show an enhanced expression of MMP-9. METHODS:Liver cryosections of the hepatocellular carcinoma(HCC)invasive front from 12 mo old Abcb4-/--mice were stained for collagen typeⅠand MMP-9 using Alexa488 and Alexa568 labeled secondary antibodies.Afterwards,the Alexa568 dye was bleached and the macrophage marker F4/80 was visualized using Alexa568 labeled secondary antibodies.Finally, photographs of the invasive tumor front were digitally overlaid and analyzed. RESULTS:After complete bleaching of the primary dye,specific fluorescence staining of a third antigen, here F4/80,was successfully performed on the same histological section.With this method,we were able to identify conglomerates of matrix metalloproteinase (MMP-9)expressing macrophages within the tumor capsule of HCC. CONCLUSION:MMP-9 expressing macrophages are involved in matrix remodelling at the invasive tumor front of HCC.The described staining protocol provides a simple yet powerful extension of conventional immuno-histochemistry,facilitating visualization of at least three different antigens plus nuclei in one single histological section.
文摘目的研究胃癌中是否存在肿瘤微环境转移(tumor microenvironment of metastasis,TMEM)及其与胃癌血道转移的相关性。方法采用免疫组化双染法同时标记巨噬细胞和内皮细胞,计数26例远处转移及其配对26例无远处转移胃癌组织中TMEM密度。结果 (1)TMEM存在于胃癌组织中;(2)TMEM与分化程度、临床分期明显相关(P<0.001,P=0.006),与肿瘤大小、Lauren分型、淋巴结转移等其他临床病理特点无关(P均>0.05);(3)TMEM与有无远处转移呈正相关(P<0.001)。结论证实胃癌中存在TMEM,TMEM密度可以预测胃癌血道转移,有望成为评估胃癌血道转移危险性的新预测指标。
