过表达SLC9A3R1对MDA-MB-231乳腺癌细胞增殖的影响

SLC9A3R1 Overexpression Inhibits Proliferation of Breast Cancer Cell Line MDA-MB-231

目的探讨SLC9A3R1在MDA-MB-231乳腺癌细胞中过表达后,对其恶性表型是否具有逆转作用。方法将pBK-CMV-HA和pBK-CMV-HA-SLC9A3R1wt质粒转染不表达SLC9A3R1的人MDA-MB-231乳腺癌细胞,经G418筛选,建立稳定高表达SLC9A3R1的MDA-MB-231细胞系。经Western blot法验证后,采用CCK-8试剂盒检测肿瘤细胞增殖活性,采用软琼脂集落形成方法检测SLC9A3R1对MDA-MB-231细胞形成集落能力的影响,并采用流式细胞仪检测SLC9A3R1对MDAMB-231细胞凋亡的影响。结果 SLC9A3R1可使MDA-MB-231细胞的增殖能力明显减弱,达50%以上(第5天P=0.0016,第6天P=0.002,第7天P=0.006)。相比MDA-MB-231细胞和整合了空载体的细胞,SLC9A3R1过表达可使MDA-MB-231细胞系锚定无关的增殖能力明显减弱[(2.9±0.47)%、(2.52±0.08)%和(1.33±0.33)%],达47%(P=0.007)。相比整合了空载体的MDA-MB-231细胞,SLC9A3R1过表达可使MDA-MB-231细胞系凋亡百分比明显增多[(2.23±1.41)%和(9.23±2.97)%],达4倍以上(P=0.018)。结论SLC9A3R1可抑制MDA-MB-231乳腺癌细胞的增殖和迁移能力。

Objective To investigate the effect of SLC9A3R1 on the proliferation, anchorage-independent growth and migration of breast cancer cell line MDA-MB-231. Methods pBK-CMV-HA-SLC9A3R1 wt vector was constructed and transfected into the tumor cell MDA-MB-231 which didn,t express SLC9A3 R.After G418 screening the cell line stably expressing SLC9A3R1 were obtained and verified by Western blot analysis. CCK-8 kit was used to detect the proliferative activity of MDA-MB-231 cell. Anchorageindependent growth was assessed by soft agar assay. FCM was applied to detect the effect of SLC9A3R1 on the apoptosis of MDA-MB-231 cell line. Results SLC9A3R1 could weaken the proliferation of MDAMB-231 cells significantly, up to 50%(P=0.0016 on day 5, P=0.002 on day 6, P=0.006 on day 7). Compared with MDA-MB-231 cells and the cells integrated of empty vectors, SLC9A3R1 overexpression could decrease the anchorage-independent growth markedly [(2.9±0.47)%,(2.52±0.08)% vs.(1.33±0.33)%], about 47%(P=0.007). Compared with MDA-MB-231 cells integrated of empty vectors, SLC9A3R1 overexpression could increase the apoptosis percent of MDA-MB-231 cell line apparently [(2.23±1.41)% vs.(9.23±2.97)%], up to 4 times(P=0.018). Conclusion SLC9A3R1 could inhibit the proliferation and migration of breast cancer cell line MDA-MB-231.