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目的探讨RNA干扰靶向抑制炭疽毒素受体1(anthrax toxin receptor 1,ANTXR1)基因的表达对食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)ECA109细胞增殖和凋亡的影响。方法利用ANTXR1siRNA转染ESCC ECA109细胞,通过实时荧光定量-聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)和Western blot检测转染ANTXR1 siRNA后ESCC ECA109细胞中ANTXR1的表达。利用细胞计数试剂盒(Cell Counting Kit-8,CCK-8)、平板克隆形成实验和流式细胞术分别检测下调ANTXR1基因的表达对ECA109细胞增殖和凋亡的影响。结果转染ANTXR1 siRNA后,ANTXR1基因在ESCC ECA109细胞中的表达明显下调(P<0.01)。CCK-8检测显示ANTXR1 siRNA组在转染3、4、5 d ECA109细胞的增殖均明显受到抑制,平板克隆形成实验显示实验组的克隆数明显低于阴性对照组;流式细胞仪检测细胞G1期细胞百分比(65.90%±6.87%)高于阴性对照组(40.17%±4.73%),组间比较差异具有统计学意义(P<0.05)。转染48 h后,实验组细胞早期凋亡率(22.61%±3.49%)显著高于对照组的早期凋亡率(8.12%±1.35%)(P<0.01)。结论 ANTXR1基因可能在ESCC ECA109细胞的增殖和凋亡中发挥重要作用,推测ANTXR1基因可能成为治疗ESCC的新靶点。
Objective To investigate the effect of RNA interference targeting the expression of anthrax toxin receptor 1 (ANTXR1) on the proliferation and apoptosis of esophageal squamous cell carcinoma (ESCC) ECA109 cells. Methods ANTXR1 siRNA was transfected into ESCC ECA109 cells and the expression of ANTXR1 in ESCC ECA109 cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Cell Counting Kit-8 (CCK-8), plate clone formation assay and flow cytometry were used to detect the effect of downregulation of ANTXR1 gene expression on the proliferation and apoptosis of ECA109 cells. Results After transfection with ANTXR1 siRNA, the expression of ANTXR1 gene was significantly down-regulated in ESCC ECA109 cells (P <0.01). The results of CCK-8 assay showed that the proliferation of ANTXR1 siRNA group was significantly inhibited at 3, 4, 5 days after transfection. The number of colonies in the experimental group was significantly lower than that in the negative control group by flow cytometry The percentage of staging cells (65.90% ± 6.87%) was higher than that of the negative control group (40.17% ± 4.73%). The difference between the two groups was statistically significant (P <0.05). At 48 h after transfection, the early apoptosis rate in the experimental group (22.61% ± 3.49%) was significantly higher than that in the control group (8.12% ± 1.35%) (P <0.01). Conclusion ANTXR1 gene may play an important role in the proliferation and apoptosis of ESCC ECA109 cells. It is speculated that ANTXR1 gene may be a new therapeutic target for ESCC.