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针对来源于乳腺癌细胞MDA-MB-435的小鼠肺癌寡灶型转移肿瘤细胞株MDA-435-OL,利用慢病毒感染的方法建立稳定敲低Sec23a基因表达的细胞株MDA-435-OL-Sec23a-GFP和其阴性对照细胞株MDA-435-OL-LV3NC-GFP,通过CCK-8(Cell Counting Kit-8)增殖实验、Transwell小室细胞迁移实验、侵袭实验和琼脂克隆斑形成实验探索敲低Sec23a基因后,寡灶型转移肿瘤细胞株体外细胞生物学特性的改变。在寡灶转移细胞株中敲低Sec23a基因后,细胞生长曲线与倍增时间并没有显著差异(28.23 h和28.32 h,P>0.05),但Transwell小室迁移细胞数量(58.50±2.81和39.60±3.21)、侵袭细胞数量(54.40±3.33和34.60±1.44)和细胞体外克隆形成率(0.67±0.05和0.37±0.03),均较阴性对照组显著增加(P<0.001)。该研究结果表明,乳腺癌寡灶型细胞株稳定敲低Sec23a基因后,细胞的增殖特性并没有明显变化,但细胞的迁移、侵袭能力和克隆形成能力均增强。
To investigate the role of MDA-435-OL in the metastasis of mouse lung cancer cell line MDA-435-OL which is derived from breast cancer cell line MDA-MB-435, the cell line stably expressing Sec23a gene was constructed by lentivirus infection. Sec23a-GFP and its negative control cell line MDA-435-OL-LV3NC-GFP were knocked down by Cell Counting Kit-8 CCK-8 proliferation assay, Transwell cell migration assay, invasion assay and agar plaque formation assay Sec23a gene, the changes of the cell biological characteristics of the tumor-bearing tumor cells in vitro. However, there was no significant difference between the cell growth curve and doubling time (28.23 h and 28.32 h, P> 0.05). However, the number of transplanted cells (58.50 ± 2.81 and 39.60 ± 3.21) , The number of invasive cells (54.40 ± 3.33 and 34.60 ± 1.44) and the rate of colony formation in vitro (0.67 ± 0.05 and 0.37 ± 0.03) were significantly increased compared with the negative control group (P <0.001). The results show that breast cancer cell line stably knocked down Sec23a gene, the proliferation of cells did not change significantly, but the cell migration, invasion and clonality were enhanced.