CAJ | 학술논문

目的观察RNA干涉后高迁移率蛋白A2(HMGA2)在垂体生长激素(GH)腺瘤细胞中表达的变化,探讨HMGA2基因表达的抑制对GH腺瘤细胞增殖、凋亡和侵袭性的影响.方法构建针对人HMGA2基因的短发夹RNA(shRNA)真核表达载体,瞬时转染GH腺瘤细胞.用Western blot法观察转染24 h后HMGA2蛋白表达水平,经细胞增殖-毒性检测试剂盒(CCK-8)测定、流式细胞仪(FCM)检测及Transwell小室检测转染后GH腺瘤细胞增殖、凋亡及侵袭.结果 HMGA2-shRNA组HMGA2蛋白表达水平与空白对照组比较下降45.58％,与scrambled组比较下降40.98％,差异有统计学意义(p＜0.05).细胞增殖反应结果显示实验组细胞增殖率显著低于对照组(P＜0.05).细胞凋亡率的结果显示与空白对照组、scrambled组比较,HMGA2-shRNA组的细胞凋亡率明显增加[(18.32±1.73)％比(7.32±1.06)％、(9.21±1.12)％,P＜0.05].体外侵袭实验显示GH腺瘤细胞侵袭力受到明显抑制(P＜0.05).结论靶向HMGA2基因的shRNA可以有效抑制垂体GH腺瘤细胞的增殖和侵袭力,并促进细胞的凋亡.
목적 관찰RNA간섭후고천이솔단백A2(HMGA2)재수체생장격소(GH)선류세포중표체적변화,탐토HMGA2기인표체적억제대GH선류세포증식、조망화침습성적영향.방법 구건침대인HMGA2기인적단발협RNA(shRNA)진핵표체재체,순시전염GH선류세포.용Western blot법관찰전염24 h후HMGA2단백표체수평,경세포증식-독성검측시제합(CCK-8)측정、류식세포의(FCM)검측급Transwell소실검측전염후GH선류세포증식、조망급침습.결과 HMGA2-shRNA조HMGA2단백표체수평여공백대조조비교하강45.58％,여scrambled조비교하강40.98％,차이유통계학의의(p＜0.05).세포증식반응결과현시실험조세포증식솔현저저우대조조(P＜0.05).세포조망솔적결과현시여공백대조조、scrambled조비교,HMGA2-shRNA조적세포조망솔명현증가[(18.32±1.73)％비(7.32±1.06)％、(9.21±1.12)％,P＜0.05].체외침습실험현시GH선류세포침습력수도명현억제(P＜0.05).결론 파향HMGA2기인적shRNA가이유효억제수체GH선류세포적증식화침습력,병촉진세포적조망.
Objective To investigate the change of high mobility protein A2 (HMGA22) after RNA interference in pituitary growth hormone (GH) adenoma cells,and explore the effects of HMGA2 gene inhibition on proliferation,apoptosis and invasion of GH adenoma cells.Methods A short hairpin RNA (shRNA)eukaryotic expression vector expressing shRNA targeting the HMGA2 gene was constructed and transfected into GH adenomas cells.The expression of HMGA2 protein was detected by using Western blotting 24 h after transfection.The growth,apoptosis and invasion of GH ademona cells were determined by using cell counting kit-8 (CCK-8),flow cytometry and Transwell assay,repectively.Results In HMGA2-shRNA group,HMGA2 protein levels were decreased bv 45.58％ and 40.98％ as compred with blank control group and scrambled group respectively (P ＜ 0.05).The proliferation rate in HMGA2-shRNA group was significantly lower than in control group (P ＜ 0.05).The apoptosis rate in HMGA2-shRNA group was significantly increased as compared with blank control group and scrambled group [(18.32 ± 1.73) ％ vs.(7.32±1.06)％ and (9.21 ±1.12)％,(P＜0.05)].The invasive ability ofGH adenoma cells was significantly inhibited in vitro (P ＜ 0.05).Conclusion ShRNA-mediated HMGA2 gene silencing can effectively inhibit proliferation and invasion of GH adenomas cells,and promote their apoptosis.