Wu Hongxue, Wang Weixing, Tong Shilun, Wu Chong
Department of Gastrointestinal, Renmin Hospital of Wuhan University Wuhan, Hubei Province, P. R. China.
Int J Clin Exp Med. 2015 Oct 15;8(10):17634-43. eCollection 2015.
The aim of this study was to investigate the effects of nucleostemin (NS) knocking down in SGC- 7901 gastric cancer cell line and investigates its correlation with the metastasis and TNM stage ingastric cancer (GC) patients.
NS expression was assessed using immunohistochemistry in 421 patients with GC. The correlation between NS expression, clinicopathological features and prognosis was analyzed. NS gene silencing was performed using a specific small interfering RNA (NS-siRNA). The gene expression level of NS was evaluated by PCR. The viability and growth rate of SGC-7901 cells were determined by trypan blue exclusion test. Cell cycle distribution of the cells was analyzed by flow cytometry.
High NS expression was correlated with node metastasis, distant metastasis and TNM stage. Kaplan-Meier survival analysis revealed that patients with low NS expression had significantly longer survival than those with high NS expression. Moreover, our results showed that NS knocking down inhibited proliferation and viability of SGC-7901 cells in a time-dependent manner. Cell cycle studies revealed that NS depletion resulted in G1 cell cycle arrest at short times of transfection (24 h) followed with apoptosis at longer times (48 and 72 h), suggest that post-G1 arrest apoptosis is occurred in SGC-7901 cells.
Overall, these results point to essential role of NS in SGC-7901 cells, thus, this gene might be considered as a promising target for treatment of GC.
本研究旨在探讨核干细胞因子(NS)敲低对SGC - 7901胃癌细胞系的影响,并研究其与胃癌(GC)患者转移及TNM分期的相关性。
采用免疫组织化学法评估421例GC患者的NS表达。分析NS表达、临床病理特征与预后之间的相关性。使用特异性小干扰RNA(NS - siRNA)进行NS基因沉默。通过PCR评估NS的基因表达水平。通过台盼蓝排斥试验测定SGC - 7901细胞的活力和生长率。通过流式细胞术分析细胞的细胞周期分布。
NS高表达与淋巴结转移、远处转移及TNM分期相关。Kaplan - Meier生存分析显示,NS低表达患者的生存期明显长于NS高表达患者。此外,我们的结果表明,NS敲低以时间依赖性方式抑制SGC - 7901细胞的增殖和活力。细胞周期研究表明,NS缺失在转染后短时间(24小时)导致G1期细胞周期阻滞,随后在较长时间(48和72小时)发生凋亡,提示SGC - 7901细胞中发生了G1期后阻滞凋亡。
总体而言,这些结果表明NS在SGC - 7901细胞中起重要作用,因此,该基因可能被视为GC治疗的一个有前景的靶点。