内源性活性氧水平及锰型超氧化物歧化酶的表达与胃癌细胞分化、增殖相关  

MAGANESE SUPEROXIDE DISMUTASE GENE TRANSCRIPTION AND ENDOGENOUS REACTIVE OXYGEN SPECIES IN DIFFERENT GASTRIC CANCER CELL LINES

作　　者：陈坚[1] 林庚金[1] 钱立平[1] 程建[2] 施冬云[2] 张亚东[2] 刘珊林[2] 

机构地区：[1]复旦大学附属华山医院消化科,上海200040 [2]复旦大学医学院生化与分子生物学系,上海200032

出　　处：《生物物理学报》2003年第2期183-188,共6页Acta Biophysica Sinica

基　　金：国家自然科学基金重点项目(30130100)

摘　　要：检测了不同分化的胃癌细胞株内MnSOD基因的表达及胞内活性氧(ROS)的水平。同时通过基因转染观察上调或下调MnSOD基因表达对SGC7901胃癌细胞胞内ROS水平及增殖能力的影响。用电穿孔法将人反义和正义MnSOD cDNA 真核表达载体pHβA-SOD (-)/pHβA-SOD (+)转入7901细胞,用含G418的RPMI1640培养基筛选稳定表达克隆。然后用RT-PCR鉴定MnDSOD基因的表达。同时用RT-PCR方法检测正常胃粘膜组织及MKN-28、SGC7901、BGC823、HGC-27四株高、中、低、未分化胃癌细胞株内的MnSOD的mRNA表达。利用DCFH-DA荧光染色方法检测不同分化胃癌细胞株及7901转染细胞株内的ROS水平。四唑蓝比色法(MTT)绘制MKN-28、SGC7901、BGC823、HGC-27四株不同分化胃癌细胞及正义、反义、空载MnSOD转染7901细胞的生长曲线。发现不同分化胃癌细胞内的MnSOD普遍呈低表达且与分化程度平行,不同分化胃癌细胞株胞内ROS水平随着MnSOD表达的下调逐步上升,细胞增殖加快。较之MnSOD空载7901,正义、反义MnSOD转染的7901细胞中该基因的表达出现明显上调及下调,胞内ROS水平较对照细胞也相应有显著降低和升高。正义株增殖受抑,反义株增殖加快。表明胃癌细胞内MnSOD的表达与肿瘤的分化程度呈负相关。可通过改变胞内ROS水平改变MnSOD基因的表达,从而调节胃癌?In order to detect the maganese superoxide dismutase(MnSOD) gene expression and the endogenous reactive oxygen species(ROS) contents in normal gastric mucosa and four gastric cancer cell lines with different degree of differentiation, semi quantitative RT-PCR and 2,7-dichlorofluorescin (DCF) assay were used respectively in the present study. The results indicate lower level of MnSOD mRNA ex-pression in MKN28 SGC7901 BGC823 and HGC-27 gastric cancer cell lines compared with that in the normal gastric mucosa while the endogenous ROS level in MKN-28?SGC7901?BGC823?HGC-27 cell lines increase synchronized with the decreasing transcription level of MnSOD in them. Furthermore, we've transfected sense or antisense MnSOD cDNA and mock carrier into SGC7901 cell line by electroporation assay. Screening stably expressed sublines by 400 mg/L G418 solution and further confirmed by RT-PCR. Evaluating the endogenous ROS level in these three sublines of SGC7901 cell transfected with sense or antisense MnSOD and vector. Evaluate the proliferative capabilities of MKN-28?SGC7901?BGC823?HGC-27 cell lines and the three sublines mentioned above via MTT assay. Sense MnSOD transfection down-regulates the endogenous ROS and inhibits the growth of SGC7901 while the antisense MnSOD transfection up-regulates the endogenous ROS and promotes the growth of SGC7901. So we conclude that MnSOD expression is related to the endogenous ROS level and differentiation degree of gastric cancer cell lines. The MnSOD gene can modulate the growth of gastric cancer cell by regulating the endogenous ROS level.

关 键 词：内源性活性氧 锰型超氧化物歧化酶 胃癌细胞 分化 增殖 

分 类 号：R735.2[医药卫生—肿瘤]