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目的通过过表达人分选连接蛋白3基因(h SNX3),以探讨h SNX3对结直肠腺癌细胞形态和迁移能力的影响。方法运用基因重组技术将h SNX3基因连接到含有GFP的慢病毒表达载体p EZ-Lv201(p EZ-SV40-e GFP-IRES-Puro)中,构建慢病毒载体p EZ-h SNX3-Lv201,感染结直肠腺癌SW620细胞,获得SNX3稳定过表达的结直肠腺癌细胞,并观察细胞形态和细胞迁移能力及相关蛋白分子的变化。结果经测序鉴定后,成功构建了慢病毒载体p EZ-h SNX3-Lv201,包装后慢病毒滴度测定为2.12×109拷贝/m L,对照慢病毒滴度为7.9×1010拷贝/m L。重组慢病毒感染SW620细胞后,经克隆筛选和Western blot法鉴定,获得了稳定过表达h SNX3的SW620h SNX3细胞。88%的SW620h SNX3细胞呈椭圆形而不是SW620原来的梭形,但TranswellTM实验和划痕试验显示,细胞迁移能力无显著变化,迁移相关蛋白上皮型钙黏素(E-cadherin)也无明显改变。结论h SNX3可改变SW620结直肠腺癌细胞的形态,但可能与细胞的迁移能力无关。
Objective To investigate the effect of h SNX3 on the morphology and migration of colorectal adenocarcinoma cells by overexpression of h SNX3. Methods The h SNX3 gene was inserted into lentiviral vector pEZ-Lv201 (pEZ-SV40-e GFP-IRES-Puro) containing GFP by gene recombination to construct lentiviral vector pEZ-h SNX3-Lv201 Colorectal adenocarcinoma SW620 cells were obtained and colorectal adenocarcinoma cells stably overexpressed by SNX3 were obtained. The morphological changes, cell migration and related molecular changes were observed. Results After sequencing, the lentiviral vector pEZ-h SNX3-Lv201 was successfully constructed. After packaging, the titer of the lentivirus was 2.12 × 109 copies / mL and the titer of the control lentivirus was 7.9 × 1010 copies / mL. The SW620h SNX3 cells stably overexpressing h SNX3 were obtained after cloned and identified by Western blot. 88% of SW620h SNX3 cells were oval rather than SW620 original spindle shape, but TranswellTM and scratch assays showed no significant changes in cell migration and no significant changes in the migration-associated protein E-cadherin . Conclusion h SNX3 can change the morphology of SW620 colorectal adenocarcinoma cells, but may not be related to cell migration ability.