目的:研究肿瘤相关抗原,黑色素瘤抗原家族A成员3(melanoma antigen family A,3;MAGE-A3),在人类膀胱癌干细胞中的表达情况,并探讨其意义。方法:采用反转录聚合酶链反应(RT-PCR)技术和Western blot技术检测MAGE-A3在人类膀胱癌细胞系T24...目的:研究肿瘤相关抗原,黑色素瘤抗原家族A成员3(melanoma antigen family A,3;MAGE-A3),在人类膀胱癌干细胞中的表达情况,并探讨其意义。方法:采用反转录聚合酶链反应(RT-PCR)技术和Western blot技术检测MAGE-A3在人类膀胱癌细胞系T24细胞中及从其中分离出来的具有干细胞特性的T24侧群细胞中的表达情况;采用免疫荧光双标记技术检测MAGE-A3和膀胱癌干细胞的一个标志物CD133在T24侧群细胞中的共表达情况。结果:在mRNA和蛋白水平,MAGE-A3在具有癌干细胞特性的T24侧群细胞中的表达水平明显高于对应的母系T24细胞;MAGE-A3和膀胱癌干细胞标志物CD133在T24侧群细胞中有阳性共表达。结论:MAGE-A3在人类膀胱癌干细胞中有特异性的高表达,有望成为膀胱癌干细胞一个新的、特异性的标志物,及针对膀胱癌干细胞进行免疫治疗的一个新的靶点。展开更多
Objective: To inwvetigate the expression of MAGE-A3 mRNA in tissue samples derived from lung cancers and to discuss the possibility of using MAGE-A3 antigens as a new peptide vaccine for inunotherapy for lung cancers...Objective: To inwvetigate the expression of MAGE-A3 mRNA in tissue samples derived from lung cancers and to discuss the possibility of using MAGE-A3 antigens as a new peptide vaccine for inunotherapy for lung cancers. Methods: Tumor tissue samples of lung cancers and paired non-tumor tissues of the lung were obtaimed from 31 lung cancer patients. Total RNA was extracted and cDNA was synthesized. Nested polymernse chain reaction amplification using MAGE-A3 specific primer was performed to detect the expression of MAGE-A3. The 10 clones of 5 samples of MAGE-A3 mRNA positive PCR products were DNA sequenced by using DNAs sequencer (PE-377). Results: Of 31 lung cancers, 26 (83.9%) expressed MACE-A3 mRNA. The expression of MAGE-A3 gene was not detectable in the adjacent lung tissues. The DNA sequencing confirmed that the target gene fragment in all 5 samples of PCR products was MACE-A3 cDNA. Point nmtations occurred in 4 samples (8 clones) detected (C^2773→T^2773; G^2807→A^2807) resulting in alternation of amino acid residue in one position (E^143→K). Conclusion: (1) The MAGE-A3 gene was expressed exclusively in tumor tissues of the patients with lung cancer in China. This tumor rejection antigen may have potential to be used as a new peptide vaccine for immunotherapy for lung eancers. (2) There are two point mutations of MAGE-A3 gene sequence in some Chinese lung cancer patients.展开更多
文摘目的:研究肿瘤相关抗原,黑色素瘤抗原家族A成员3(melanoma antigen family A,3;MAGE-A3),在人类膀胱癌干细胞中的表达情况,并探讨其意义。方法:采用反转录聚合酶链反应(RT-PCR)技术和Western blot技术检测MAGE-A3在人类膀胱癌细胞系T24细胞中及从其中分离出来的具有干细胞特性的T24侧群细胞中的表达情况;采用免疫荧光双标记技术检测MAGE-A3和膀胱癌干细胞的一个标志物CD133在T24侧群细胞中的共表达情况。结果:在mRNA和蛋白水平,MAGE-A3在具有癌干细胞特性的T24侧群细胞中的表达水平明显高于对应的母系T24细胞;MAGE-A3和膀胱癌干细胞标志物CD133在T24侧群细胞中有阳性共表达。结论:MAGE-A3在人类膀胱癌干细胞中有特异性的高表达,有望成为膀胱癌干细胞一个新的、特异性的标志物,及针对膀胱癌干细胞进行免疫治疗的一个新的靶点。
基金This project was supported by the key project of Scientific Committee of Henan Province (No. 0124170232), the key project ofZhengzhou Scientific Committee (No. 04BA60ABYD18), and Tumor Biology Subproject of 211 project of zhengzhou Univevsity
文摘Objective: To inwvetigate the expression of MAGE-A3 mRNA in tissue samples derived from lung cancers and to discuss the possibility of using MAGE-A3 antigens as a new peptide vaccine for inunotherapy for lung cancers. Methods: Tumor tissue samples of lung cancers and paired non-tumor tissues of the lung were obtaimed from 31 lung cancer patients. Total RNA was extracted and cDNA was synthesized. Nested polymernse chain reaction amplification using MAGE-A3 specific primer was performed to detect the expression of MAGE-A3. The 10 clones of 5 samples of MAGE-A3 mRNA positive PCR products were DNA sequenced by using DNAs sequencer (PE-377). Results: Of 31 lung cancers, 26 (83.9%) expressed MACE-A3 mRNA. The expression of MAGE-A3 gene was not detectable in the adjacent lung tissues. The DNA sequencing confirmed that the target gene fragment in all 5 samples of PCR products was MACE-A3 cDNA. Point nmtations occurred in 4 samples (8 clones) detected (C^2773→T^2773; G^2807→A^2807) resulting in alternation of amino acid residue in one position (E^143→K). Conclusion: (1) The MAGE-A3 gene was expressed exclusively in tumor tissues of the patients with lung cancer in China. This tumor rejection antigen may have potential to be used as a new peptide vaccine for immunotherapy for lung eancers. (2) There are two point mutations of MAGE-A3 gene sequence in some Chinese lung cancer patients.