Zhang Jing, Fu Yan-chao, Kang Chun-sheng, Zhang Qing-yu, Wang Tao, Zhang Jie
Department of Gastroenterology, Tianjin Medical University General Hospital, China.
Zhonghua Yi Xue Za Zhi. 2009 Aug 25;89(32):2292-5.
To construct a short hairpin RNA (shRNA) adenovirus vector targeting Akt1 (protein kinase B1, PKB1/Akt1) and cyclooxygenase-2 (COX-2) and study its effects on the growth of SGC-7901 human gastric adenocarcinoma cell.
Adenovirus pGSadeno-Akt1 + 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 rAd5-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.
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 no-load 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.
Adenovirus-mediated Akt1 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.
构建靶向Akt1（蛋白激酶B1，PKB1/Akt1）和环氧化酶-2（COX-2）的短发夹RNA（shRNA）腺病毒载体，并研究其对人胃腺癌SGC-7901细胞生长的影响。
构建腺病毒pGSadeno-Akt1 + COX-2（rAd5-A + C）载体并转染至SGC-7901细胞。采用MTT法和流式细胞术在体外评估肿瘤细胞的增殖活性。将rAd5-HK和rAd5-A + C注射到已建立的裸鼠皮下SGC-7901胃腺癌模型中。在21天的观察期内，每3天测量一次肿瘤体积，以进一步观察rAd5-A + C对SGC-7901细胞的抗肿瘤作用，并通过TUNEL法检测细胞原位凋亡情况。
将构建的腺病毒载体rAd5-A + C转染至SGC-7901细胞后，rAd5-A + C处理组的细胞增殖活性明显受到抑制，细胞周期分析表明，对照组SGC-7901和空载组rAd5-A + C细胞在G0/G1期、S期和G2/M期的细胞分别占细胞总数的49.8%、35.2%、15.0%和50.8%、36.5%、12.7%。而处理组rAd5-A + C在G0/G1期、S期和G2/M期的细胞分别占细胞总数的68.1%、21.8%和10.1%。处理组的肿瘤体积低于对照组和空载组，差异具有统计学意义（F = 3.679，P = 0.043），且rAd5-A + C可诱导肿瘤细胞凋亡。
腺病毒介导的Akt1和COX-2 shRNA可抑制人胃腺癌SGC-7901细胞的生长。这可能为胃癌基因治疗提供一种新策略。
