Hu Yang, Pu Qingha, Cui Bin, Lin Jia
Department of General Surgery, Sichuan Academy of Medical Science & Sichuan Provincial People's Hospital Chengdu 610072, China.
Department of General Surgery, The First Affiliated Hospital of Chongqing Medical University Chongqing 400016, China.
Int J Clin Exp Pathol. 2015 Aug 1;8(8):8921-8. eCollection 2015.
To study how miR-34a acts as a tumor suppressor in inhibiting the invasion and metastasis of the gastric cancer cells. First, real-time polymerase chain reaction (PCR) and western blot analysis were used to analyze the expression of miR-34a and Tgif2 in gastric cancer tissues and the adjacent normal tissues. Next, gastric cancer cells were transfected with miR-34a mimic and Tgif2 siRNA, respectively. After transfection, real-time PCR and western blot analysis were used to detect the relative Tgif2 expression level. Cell proliferation was monitored by the colorimetric water-soluble tetrazolium salt and apoptosis analysis was performed with Annexin-V-FITC Apoptosis Detection Kit I. The expression of miR-34a in the adjacent non-tumor tissues was higher than that in gastric cancer tissues, but Tgif2 was opposite. In gastric cancer cells transfected with miR-34a mimic/Tgif siRNA, Tgif2 expression was remarkably down-regulated. Cells transfected with miR-34a mimic/Tgif2 siRNA grew more slowly than the control groups. The percentage of apoptotic cells in gastric cancer cells transfected with miR-34a mimic/Tgif siRNA was much higher compared to the controls. Therefore, we concluded that miR-34a could inhibit tumor invasion and metastasis in gastric cancer by targeting Tgif2 and may be a novel therapeutic candidate for gastric cancer.
为研究miR-34a作为肿瘤抑制因子在抑制胃癌细胞侵袭和转移中的作用机制。首先，采用实时聚合酶链反应（PCR）和蛋白质免疫印迹分析来检测胃癌组织及癌旁正常组织中miR-34a和Tgif2的表达。接着，分别用miR-34a模拟物和Tgif2小干扰RNA（siRNA）转染胃癌细胞。转染后，采用实时PCR和蛋白质免疫印迹分析检测Tgif2的相对表达水平。通过比色法水溶性四氮唑盐监测细胞增殖，并使用膜联蛋白V-异硫氰酸荧光素凋亡检测试剂盒I进行凋亡分析。癌旁非肿瘤组织中miR-34a的表达高于胃癌组织，而Tgif2的表达情况则相反。在转染了miR-34a模拟物/Tgif2 siRNA的胃癌细胞中，Tgif2的表达显著下调。转染了miR-34a模拟物/Tgif2 siRNA的细胞比对照组生长得更慢。与对照组相比，转染了miR-34a模拟物/Tgif2 siRNA的胃癌细胞中凋亡细胞的百分比要高得多。因此，我们得出结论，miR-34a可通过靶向Tgif2抑制胃癌的肿瘤侵袭和转移，可能是一种新型的胃癌治疗候选药物。
