增殖诱导配体shRNA对结直肠癌细胞增殖与转移的影响

目的 探讨增殖诱导配体(APRIL)shRNA对结直肠癌细胞的增殖与转移的影响.方法 用RNA干扰技术敲低结直肠癌细胞株SW480中APRIL基因的表达,以免疫印迹技术、细胞黏附、损伤修复及侵袭力检测等体外实验对比APRIL基因敲低前后细胞株SW480的增殖、黏附、侵袭及转移能力的变化.结果 转染组细胞APRIL蛋白表达的灰度值为0,显著低于阴性对照组的1.0±0.3和未转染组的1.1±0.4,差异有统计学意义(F=42.15,P=0.00);转染组、阴性对照组及未转染组最大增殖细胞数分别为(6.1±4.7)×104个、(36.4±14.4)×104个和(41.3±19.2)×104个,转染组细胞增殖能力明显受到抑制(F=24.76,P<0.05);转染组细胞黏附的平均吸光度(A)值为0.343,低于阴性对照组的0.409及未转染组的0.412,细胞黏附率相对于未转染组降低39%;转染组、阴性对照组及未转染组迁移至损伤区细胞数分别为(47.89±13.16)个、(98.78±23.26)个和(108.01±39.11)个,与阴性对照组、未转染组比较,细胞迁移能力的差异有统计学意义(F=65.53,P<0.01);细胞侵袭力(A值0.58±0.10)相对于阴性对照组(A值0.94±0.23)与未转染组(A值1.11±0.21)差异亦有统计学意义(F=5.771,P<0.05).结论 APRIL有促细胞增殖作用,很可能参与结直肠癌细胞的侵袭和转移,并具增强结直肠癌侵袭和转移潜能.

Short hairpin RNA-mediated silencing of APRIL gene inhibits proliferation and metastasis of human colon carcinoma SW480 cells in vitro

Objective To study the influence of short hairpin RNA-mediated inhibition of APRIL gene on proliferation,invasion and metastasis of human colon carcinoma SW480 ceils in vitro. Methods After treatment with shRNA, the expression level of APRIL protein in human colon carcinoma SW480 cells was detected by western blotting. And cell adhesion, cell migration and cell invasion of SW480 cells transfected with sh637 were analyzed respectively as compared with non-transfected SW480 cells and mocktransfectant. Results The expression of APRIL protein in SW480 transfected with sh637 were significantly lower than mock-plasmid groups and untransfected ones (F=42.15, P=0.00). Cell proliferation was markedly inhibited, compared with untransfected groups and negative control ones (F=24.76, P<0.05).And the average absorption of cell adhesion in transfected groups, mock-plasmid and non-transfected ones were 0.343, 0.409, 0.412, respectively. Cell adhesion decreased 39% compared with untransfected groups. Similarly, there was significant difference for cell migration (F=65.53, P<0.01) between transfected groups (The average ceil number in exposed area to migrate was 47.89±13.16) and nontransfected (98.78±23.26) as well as mock-plasmid ones (108.01±39.11). Then, in view of the average absorption of cell invasion between transfected groups(0.58±0.10) and non-transfected (0.94±0.23) as well as mock-plasmid ones(1.11±0.21), a prominently difference for cell invasion was also found between them (F=5.771, P<0.05). Conclusion The results suggest APRIL has a proliferation-inducing effect and probably be involved in cell invasion and cell metastasis of colon carcinoma and it could enhance capacity of invasion and metastasis of SW480 cells.