Liu Ping, Wang Tong-shan, You Si-hong, Ge Hong-Mei
Department of Oncology, the First Affiliated Hospital, Nanjing Medical University, Nanjing 210029, China.
Zhonghua Zhong Liu Za Zhi. 2007 Aug;29(8):570-4.
The aim of this study was to detect the expression of livin in human gastric carcinoma and analyze the relationship between livin expression and cliniopathologic features. To explore the feasibility of small interference RNA (siRNA) in inhibition of livin gene expression and to investigate the apoptosis susceptibility of SGC-7901 cells by siRNA-mediated silencing of the livin gene.
The expression of livin at mRNA and protein levels were determined by RT-PCR and Western blot assay, respectively. The relationship between livin expression and clinicopathologic features was analyzed. Two siRNAs specifically targeting livin gene were designed and synthesized in vitro, and were transfected into the gastric cancer SGC-7901 cells. The expression of livin mRNA was assayed by RT-PCR. Cell growth state and 50% inhibition concentration (IC50) of 5-Fu and cisplatin on SGC-7901 cells were determined by MTT method. Cell apoptosis was assessed by flow cytometry (FCM).
The expressions of livin mRNA and protein were detected in 19 of 40 gastric carcinoma cases (47.5%). No expression of livin was detected in tumor-adjacent tissues and benign gastric lesion. A positive correlation was found between livin expression and poor differentiation as well as lymph node metastases (P < 0.05). The level of livin mRNA was decreased in the SGC-7901 cells transfected by si-livin1 for 48 hours, with inhibition of cell growth. IC50 of si-livin-treated SGC-7901 cells to 5-Fu and cisplatin was decreased (P < 0.05) and the cells were more susceptible to proapoptotic stimuli (5-Fu and cisplatin) than control groups (P < 0.05).
Livin is overexpressed in gastric carcinoma with a correlation to tumor differentiation and lymph node metastasis, suggesting that it may be as one of molecular prognostic factors for some cases of gastric cancer. SiRNA can inhibit livin expression of SGC-7901 cells and induce cell apoptosis. Livin might serve as a new target for apoptosis-inducing therapy of gastric cancer. Livin;
本研究旨在检测人胃癌组织中生存素(livin)的表达情况,分析其表达与临床病理特征之间的关系。探讨小干扰RNA(siRNA)抑制livin基因表达的可行性,以及通过siRNA介导沉默livin基因来研究SGC - 7901细胞的凋亡敏感性。
分别采用逆转录聚合酶链反应(RT - PCR)和蛋白质免疫印迹法(Western blot)检测livin在mRNA和蛋白水平的表达。分析livin表达与临床病理特征之间的关系。体外设计并合成两条特异性靶向livin基因的siRNA,转染胃癌SGC - 7901细胞。采用RT - PCR检测livin mRNA的表达。通过MTT法测定细胞生长状态以及5 - 氟尿嘧啶(5 - Fu)和顺铂对SGC - 7901细胞的半数抑制浓度(IC50)。采用流式细胞术(FCM)评估细胞凋亡情况。
40例胃癌组织中有19例(47.5%)检测到livin mRNA和蛋白表达。癌旁组织及胃良性病变组织中未检测到livin表达。livin表达与低分化以及淋巴结转移呈正相关(P < 0.05)。转染si - livin1 48小时后,SGC - 7901细胞中livin mRNA水平降低,细胞生长受到抑制。si - livin处理的SGC - 7901细胞对5 - Fu和顺铂的IC50降低(P < 0.05),且与对照组相比,细胞对促凋亡刺激因素(5 - Fu和顺铂)更敏感(P < 0.05)。
生存素在胃癌中呈过表达,与肿瘤分化及淋巴结转移相关,提示其可能作为部分胃癌病例的分子预后指标之一。SiRNA可抑制SGC - 7901细胞中livin的表达并诱导细胞凋亡。生存素可能成为胃癌诱导凋亡治疗的新靶点。生存素;