靶向蛋白激酶B1和环氧合酶2短发夹RNA对胃腺癌细胞生长的作用  

Inhibitory effects of targeting protein kinase B1 and cyclooxygenase-2 shRNA upon human gastric adenocarcinoma cell growth

作　　者：张靖[1] 付彦超[1] 康春生[1] 张庆瑜[1] 王涛[1] 张洁[1] 

机构地区：[1]天津医科大学总医院消化科,300052

出　　处：《中华医学杂志》2009年第32期2292-2295,共4页National Medical Journal of China

基　　金：基金项目：天津市应用基础重点项目（07JCZDJC07700）

摘　　要：目的研究靶向蛋白激酶B1（PKB1／Akt1）和环氧合酶2（COX-2）短发夹RNA（shRNA）腺病毒载体对SGC-7901人胃腺癌细胞生长的作用。方法构建腺病毒载体pGSadeno—Akt1＋COX-2（rAd5-A＋C），体外转染人胃癌细胞株SGC-790／后，噻唑蓝比色分析法（MTT法）和流式细胞术评价转染后胃癌细胞的增殖活性。裸鼠皮下荷瘤模型进--步观察，Ad5-A＋C对SGC-7901生长的抑制效果，并应用原位末端标记技术检测肿瘤细胞的原位凋亡。结果构建的rAd5-A＋C重组腺病毒载体转染SGC-7901后，胃癌细胞的增殖活性明显被抑制。细胞周期分析显示对照组和空载组在G0／G1期，S期和G2／M期的细胞数占细胞总数分别为49．8％、35．2％、15．0％和50．8％、36．5％、12．7％；而治疗组在G0／G1期、S期和G2／M期的细胞数占细胞总数的68．1％、21．8％和10．1％。裸鼠皮下荷瘤模型显示rAd5-A＋C可抑制肿瘤的生长和诱导细胞的凋亡。结论腺病毒介导的Akt1和COX-2shRNA可抑制人胃癌细胞的生长，这可能为胃癌靶向性联合基因治疗提供了新的策略。Objective To construct a short hairpin RNA （shRNA） adcnovirus vector targeting Aktl （protein kinase B1, PKBI/Aktl） and cyclooxygenase-2 （COX-2） and study its effects on the growth of SGC-7901 human gastric adenocarcinoma cell. Methods Adenovirus pGSadeno-Aktl ＋ COX-2 （rAd5-A ＋ C） vector was constructed and transfected into SGC-7901 cell. The proliferative activity of tumor cell was evaluated by MTT assay and flow cytometry in vitro, rAd5-HK and rAdS-A ＋ C were injected into the established subcutaneous SGC-7901 gastric adenocarcinoma in nude mice. During the observation period of 21 days, tumor volume was measured every 3 days to further observe the anti-tumor effects of rAd5-A ＋ C on SGC-7901 cell and cell situ apoptosis was detected by TUNEL assay. Results After transfection of constructed adenovirus vector rAd5-A ＋ C into SGC-7901 cell, cell proliferative activity in rAd5-A ＋ C treatment group was significantly suppressed, and cell cycle indicated that control group SGC-7901 and noload group rAd5-A ＋ C cells in G0/G1 ,S and G2/M phases accounted for the total number of cells 49. 8% , 35.2%, 15.0% and 50. 8%, 36. 5% , 12. 7% respectively. While the treatment group rAd5-A ＋ C in G0/ G1 ,S and G2/M phases accounted for the total number of cells 68. 1% , 21.8% and 10. 1% respectively. The tumor volume in treatment group was lower than that of control and no-load groups and the difference had statistical significance （ F =3. 679, P = 0. 043 ） and rAd5-A ＋ C could induce the apoptosis of tumor cell. Conclusion Adenovirus-mediated Aktl and COX-2 shRNA can inhibit the growth of SGC-7901 human gastric adenocarcinoma cell. It may provide a new strategy for gastric cancer gene therapy.

关 键 词：蛋白激酶类 环氧化酶2 胃肿瘤 腺病毒载体 

分 类 号：R686[医药卫生—骨科学]