Zhang Y, Chen Y, Qu C, Zhou M, Ni Q, Xu L
Xinhua Hospital, School of Medicine, Shanghai Jiaotong University Department of Digestive Endoscopy Shanghai China.
Xinhua Hospital, School of Medicine, Shanghai Jiaotong University Department of Digestive Endoscopy Shanghai China lming_xu@163.com.
Cell Mol Biol (Noisy-le-grand). 2014 Mar 25;60(1):26-32.
In the present study we investigate the effects of small interfering RNA (siRNA)-mediated silencing of prohibitins (Phbs) gene expressions on gastric carcinoma cell proliferation and apoptosis in vitro and in vivo. Firstly, Phbs proteins in five human gastric carcinoma cell lines were evaluated by Western blotting. Then three siRNA sequences targeting Phbs were designed and transfected into the cancer cell line with the highest Phbs protein expressions. The cell proliferation and apoptosis were analyzed by MTT and flow cytometry methods. In vivo, the effects of siRNAs on the proliferation and apoptosis were assayed in xenografted tumors of nude mice. Phbs proteins were expressed highest in gastric carcinoma cell line SGC7901. The cell proliferation significantly decreased upon Phbs silencing. The cell percentage in S phase was significantly lower in Phbs silenced groups, and the apoptotic cell ratio was significantly higher. siRNA targeting Phbs significantly inhibited the growth of the xenografted tumors. Further, Bcl-2 and Bax was respectively downregulated and upregulated after the Phbs silencing in the xenografted tumors. Caspase-3 and caspase-9 activities were significantly enhanced in the Phbs-silenced tumors. In summary, siRNA-mediated silencings of Phbs inhibit proliferation and promotes apoptosis of gastric carcinoma cells SGC7901 in vitro and in vivo, suggesting key roles of Phbs in the development of SGC7901 cell line.
在本研究中，我们调查了小干扰RNA（siRNA）介导的抑制素（Phbs）基因表达沉默对胃癌细胞体外和体内增殖及凋亡的影响。首先，通过蛋白质印迹法评估五种人胃癌细胞系中的Phbs蛋白。然后设计了三个靶向Phbs的siRNA序列，并将其转染到Phbs蛋白表达最高的癌细胞系中。通过MTT法和流式细胞术分析细胞增殖和凋亡情况。在体内，在裸鼠异种移植瘤中测定siRNA对增殖和凋亡的影响。Phbs蛋白在胃癌细胞系SGC7901中表达最高。Phbs沉默后细胞增殖显著降低。Phbs沉默组中S期细胞百分比显著降低，凋亡细胞比例显著升高。靶向Phbs的siRNA显著抑制异种移植瘤的生长。此外，在异种移植瘤中Phbs沉默后，Bcl-2下调，Bax上调。Phbs沉默的肿瘤中Caspase-3和Caspase-9活性显著增强。总之，siRNA介导的Phbs沉默在体外和体内均抑制胃癌细胞SGC7901的增殖并促进其凋亡，提示Phbs在SGC7901细胞系发育中起关键作用。
