长链非编码RNA母系表达基因3调节肾透明细胞癌增殖和凋亡的功能研究  

Role of long non - coding RNA maternally expressed gene 3 on cell proliferation and apoptosis of clear cell renal cell carcinoma

作　　者：程鹤鹏[1] 李扬[1] 李晓东[1] 刘骞[1] 张广伟[1] 朱朝阳[1] 

机构地区：[1]河南大学淮河医院泌尿外科,开封475000

出　　处：《中华实验外科杂志》2016年第6期1468-1470,共3页Chinese Journal of Experimental Surgery

基　　金：国家高技术研究发展计划（863计划：2014AA020608）

摘　　要：目的观察长链非编码RNA（1ncRNA）母系表达基因3（MEG3）在人肾透明细胞癌组织和细胞株中的表达，探讨过表达MEG3后对细胞增殖和凋亡的影响。方法采用实时定量聚合酶链反应（Real—timePCR）检测MEG3在32对配对人肾透明细胞癌组织和癌旁症状组织及6株人肾透明细胞癌细胞株中的表达水平；在786-O细胞中稳定转染MEG3过表达质粒，分别利用噻唑蓝（MTF）实验和流式细胞实验分析MEG3过表达后对细胞增殖和凋亡的影响；Westernblot检测增殖与凋亡相关分子标志物。结果肾透明细胞癌组织和细胞株中MEG3的表达明显低于对应的正常组织和细胞株；空载pcDNA3．1细胞与稳定转染MEG3的pcDNA3．1-MEG3细胞MEG3表达水平分别为0．51±0．02和11．26±1．89，差异有统计学意义（P〈0．05）；MTF实验结果显示，MEG3过表达后786-O细胞的增殖活性显著降低（P〈0．05）；流式细胞术检测细胞凋亡比例，空载pcDNA3．1细胞与稳定转染MEG3的pcDNA3．1-MEG3细胞凋亡比例分别为（7．02±1．21）％和（13．95±5．48）％，差异有统计学意义（P〈0．05）；两株细胞株中MEG3的过表达均可导致细胞周期素D1（CyclinD1）下降和促凋亡蛋白B细胞淋巴瘤／白血病-2相关X蛋白（bax）上升。结论肾透明细胞癌中MEG3表达降低；MEG3可通过影响细胞增殖和凋亡在肾透明细胞癌中发挥抑癌作用。Objective To examine the expression of long non -coding RNA maternally expressed gene 3 （ MEG3 ） in human clear cell renal cell carcinoma tissues （ccRCC） and cell lines. And to analyze the effects of MEG3 overexpression on the proliferation and apoptosis of ecRCC cells. Methods Real - time quantitative polymerase chain reaction （ Real - time PCR） was carried out to measure the expression level of MEG3 in ccRCC specimens and cell lines compared with respective controls. After MEG3 was sta- bilized overexpressed in 786 -O cells, methyl thiazol tetrazolium （MTF） and flow- cytometric assays were performed to evaluate the effects of MEG3 on cell proliferation and apoptosis. Protein levels of growth - or apoptosis- related molecules were then determined by Western blotting. Results MEG3 expression was decreased in ccRCC specimens and cell lines compared with respective controls. Overexpression of MEG3 in ccRCC cell line 786 - O [0. 51 ＋0. 02 vs. 11.26 ＋ 1.89 for pcDNA3.1 and pcDNA3.1 - MEG3 cell lie respectively with statistical difference （ P 〈 0. 05 ） ] resulted in delayed cell proliferation rate and induction of cell apoptosis[ （7.02 ＋ 1.21 ）% vs. （ 13.95 ＋ 5.48 ）% ] in vitro. Cyelin D1 and B cell lymphoma/leu- kemia - 2 associated X protein （bax） protein levels were affected by MEG3 overexpression in these cell lines. Conclusion MEG3 was down- regulated in ccRCC, in which it functions as a potent tumor sup- pressor by regulating cell proliferation and apoptosis processes.

关 键 词：长链非编码RNA 母系表达基因3 肾透明细胞癌 细胞增殖 凋亡 

分 类 号：R737.11[医药卫生—肿瘤]