High mobility group A2（HMGA2） protein is a small nonhistone chromosomal protein that can modulate transcription of an ample number of genes.Many previous studies demonstrate that up-regulation of HMGA2 expression oc...High mobility group A2（HMGA2） protein is a small nonhistone chromosomal protein that can modulate transcription of an ample number of genes.Many previous studies demonstrate that up-regulation of HMGA2 expression occurrs in many kinds of cancers including colorectal cancer,suggesting that HMGA2 might play a critical role in the progression of various tumors.However,the exact role of HMGA2 in colorectal cancer has not been determined.To verify the essential role of HMGA2 in the growth and invasiveness of colorectal cancer,HMGA2 expression was down-regulated by RNA interference（RNAi） in SW480 cells.We observed that the knockdown of HMGA2 led to the significant inhibition of proliferation and invasion of SW480 cells in vitro.These results suggest that HMGA2 might play a crucial role in the progression of colorectal cancer,and be a potential therapeutic target for human colorectal cancer.展开更多
目的用分子生物学研究肾癌形成和进展,可为临床治疗提供更可靠的依据,高迁移率族蛋白A2(high mobility group A2,HMGA2)是近年来研究热点之一。本研究观察HMGA2对肾癌细胞增殖和侵袭能力影响,为肾癌进一步临床研究提供依据。方法培养人...目的用分子生物学研究肾癌形成和进展,可为临床治疗提供更可靠的依据,高迁移率族蛋白A2(high mobility group A2,HMGA2)是近年来研究热点之一。本研究观察HMGA2对肾癌细胞增殖和侵袭能力影响,为肾癌进一步临床研究提供依据。方法培养人肾癌细胞系786-O、769-P、人肾癌转移细胞系ACHN、人正常肾小管上皮细胞系HKC,研究分为3组,HMGA2-siRNA组、Mock-siRNA组和未转染组。利用RNA干扰技术,瞬时转染肾癌ACHN细胞,采用逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)和蛋白质印迹法检测HMGA2mRNA及蛋白表达;MTT法检测干扰后ACHN细胞增殖能力;Transwell法检测干扰后ACHN细胞侵袭能力。结果786-O、769-P、ACHN和HKC细胞系HMGA2mRNA表达量分别为0.82±0.10、0.79±0.09、0.98±0.16和0.04±0.01,组内差异有统计学意义,F=63.36,P=0.002;4种细胞系中HMGA2蛋白表达量分别为0.80±0.09、0.75±0.08、0.94±0.10和0.06±0.02,组内差异有统计学意义,F=49.09,P=0.005。选择ACHN细胞作为后续研究:成功构建特异性沉默HMGA2表达的肾癌细胞株。在转染后24、48、72和96h,HMGA2-siRNA组细胞增殖速度分别为0.69±0.02、0.90±0.05、0.99±0.07和1.10±0.09,Mock-siRNA组细胞增殖速度分别为0.89±0.03、1.36±0.06、1.82±0.08和2.10±0.10,未转染组细胞增殖速度分别为0.91±0.02、1.50±0.06、1.91±0.10和2.20±0.12。HMGA2-siRNA组细胞增殖速度低于Mock-siRNA组和未转染组,F组别=630.724,P<0.001;F时间=566.968,P<0.001;F组别×时间=51.328,P=0.002。在转染后48h,HMGA2-siRNA组、Mock-siRNA组及未转染组穿过细胞数的比值分别为0.34±0.04,0.87±0.09和0.91±0.10,F=6.42,P=0.039。结论用HMGA2-siRNA干扰ACHN细胞,可抑制细胞增殖及侵袭能力,HMGA2基因在肾癌发生、发展中可能发挥“癌基因”作用,HMGA2基因及蛋白表达可能与肿瘤形成、进展和转移密切相关,有望成为肾癌治疗的一个重要靶点。展开更多
文摘High mobility group A2（HMGA2） protein is a small nonhistone chromosomal protein that can modulate transcription of an ample number of genes.Many previous studies demonstrate that up-regulation of HMGA2 expression occurrs in many kinds of cancers including colorectal cancer,suggesting that HMGA2 might play a critical role in the progression of various tumors.However,the exact role of HMGA2 in colorectal cancer has not been determined.To verify the essential role of HMGA2 in the growth and invasiveness of colorectal cancer,HMGA2 expression was down-regulated by RNA interference（RNAi） in SW480 cells.We observed that the knockdown of HMGA2 led to the significant inhibition of proliferation and invasion of SW480 cells in vitro.These results suggest that HMGA2 might play a crucial role in the progression of colorectal cancer,and be a potential therapeutic target for human colorectal cancer.
