Zhang Yan, Zhang Meiwu, Fan Xiaoxiang, Mao Dafeng, Lv Shuyi, Chen Ping
Department of Interventional Therapy, Ningbo No. 2 Hospital, Ningbo, Zhejiang 315010, P.R. China.
Department of Gastrointestinal and Minimally Invasive Surgery, Ningbo No. 2 Hospital, Ningbo, Zhejiang 315010, P.R. China.
Oncol Lett. 2019 Feb;17(2):2151-2158. doi: 10.3892/ol.2018.9814. Epub 2018 Dec 10.
Effect of STAT3 decoy oligodeoxynucleotides (ODN) transduced by ultrasound microbubbles combined with ultrasound on the growth of esophageal squamous cell carcinoma and its mechanism were analyzed. EC9706 cells were cultured in vitro and divided into four groups: group E (ultrasound microbubble + ultrasound irradiation), group P (liposome + ultrasound irradiation), group C (ultrasound), and group CC (ultrasound microbubbles). Mutant ODNs were used in groups E and P and the control group was group EC and PC, respectively. Immunofluorescence assay and flow cytometry were used to detect the transfection efficiency of each group. MTT colorimetric assay was performed to analyze the inhibition rate in each group. The effect of STAT3 decoy ODN on the proliferation of esophageal squamous carcinoma cells was calculated. Revese transcription-quantitative PCR (RT-qPCR) and western blotting were performed to detect the expression of the STAT signaling pathway downstream of gene expression levels. The model of subcutaneous transplantation of nude mice was used to show the effect of different transfections and STAT3 decoy ODN on the weight and volume of the transplanted tumor in mice. The cell inhibition rate was higher in group E than in groups P (F=8.382, P<0.001) and CC (F=6.469, P<0.001). Compared with groups EC, PC and C, respectively, the mRNA expression of STAT3, bcl-xL and Cyclin D1 decreased in groups E, P and CC (F=5.328, P<0.001). The weight and volume of nude mice in groups E, P and CC exhibited an inhibitory effect on the weight and volume of nude mice. Ultrasound irradiation combined with ultrasound microbubbles is an effective transfection method. The transfection of STAT3 decoy ODN can significantly inhibit the activity of esophageal squamous cell carcinoma cells and enhance apoptosis of cells, which has potential clinical value.
分析超声微泡联合超声转导的信号转导和转录激活因子3（STAT3）诱饵寡脱氧核苷酸（ODN）对食管鳞状细胞癌生长的影响及其机制。将EC9706细胞体外培养并分为四组：E组（超声微泡+超声照射）、P组（脂质体+超声照射）、C组（超声）和CC组（超声微泡）。E组和P组使用突变ODN，对照组分别为EC组和PC组。采用免疫荧光分析和流式细胞术检测各组的转染效率。采用MTT比色法分析各组的抑制率。计算STAT3诱饵ODN对食管鳞癌细胞增殖的影响。采用逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测STAT信号通路下游基因表达水平的表达。采用裸鼠皮下移植模型观察不同转染和STAT3诱饵ODN对小鼠移植瘤重量和体积的影响。E组细胞抑制率高于P组（F=8.382，P<0.001）和CC组（F=6.469，P<0.001）。与EC组、PC组和C组相比，E组、P组和CC组中STAT3、bcl-xL和细胞周期蛋白D1的mRNA表达均降低（F=5.328，P<0.001）。E组、P组和CC组裸鼠的体重和体积对裸鼠的体重和体积有抑制作用。超声照射联合超声微泡是一种有效的转染方法。STAT3诱饵ODN转染可显著抑制食管鳞状细胞癌细胞活性并增强细胞凋亡，具有潜在临床价值。
