EDN3基因抑制人乳腺癌细胞的迁移及侵袭

EDN3 gene inhibits the migration and invasion of breast cancer cells

目的 :探讨内皮素3(endothelin 3,EDN3)基因对人乳腺癌细胞迁移及侵袭能力的影响。方法 :应用RT-PCR法检测EDN3及内皮素受体B(endothelin receptor B,EDNRB)m RNA在人乳腺癌细胞(BT549、MB231、MDA-MB468、MCF-7、SK-BR-3、T47D和BT474)中的表达以及EDN3 m RNA在正常乳腺组织中的表达情况。应用实时荧光定量PCR法检测EDN3 m RNA在人乳腺癌组织及对应的癌旁组织中的表达情况。构建表达EDN3基因的慢病毒重组载体p CDH-CMV-MCS-EF1-Puro-EDN3,病毒包装后将其感染乳腺癌BT549和MB231细胞,应用CCK-8法及Transwell小室法分别检测EDN3基因对乳腺癌细胞增殖、迁移及侵袭能力的影响。结果 :EDN3 m RNA在人乳腺癌细胞中表达缺失,而在乳腺正常组织中可见表达。EDNRB mR NA在BT549、MB231、T47D和SK-BR-3细胞中表达。乳腺癌组织中EDN3 mR NA的表达水平低于对应的癌旁组织(P=0.000 6)。成功构建表达EDN3基因的重组慢病毒载体,并将其成功感染BT549和MB231细胞。EDN3表达对BT549和MB231细胞的增殖无显著影响(P值均>0.05),但能抑制其迁移和侵袭(P值均<0.01)。结论 :人乳腺癌中EDN3基因的表达显著下调。EDN3基因可抑制人乳腺癌细胞的迁移及侵袭。

Objective： To investigate the effect of endothelin 3 （EDN3） gene on the migration and invasion abilities of human breast cancer cells. Methods： The expressions of EDN3 and endothelin receptor B （EDNRB） mRNAs in breast cancer cell lines BT549, MB231, MDA-MB468, MCF-7, SK-BR-3, T47D and BT474 and the expression of EDN3 mRNA in human normal breast cancer tissues were detected by RT- PCR, The expression of EDN3 mRNA in breast cancer tissues and the corresponding para-cancerous tissues was detected by real-time fluorescent quantitative PCR. Recombinant lentivirus vector pCDH-CMV-MCS-EF1-Puro-EDN3 expressing EDN3 gene was constructed. After recombinant lentivirus was packaged and infected into breast cancer BT549 and MB23t cells, the abilities of proliferation, migration and invasion of breast cancer cells were detected by CCK-8 and Transwell assay. Results： The expression of EDN3 mRNA in human breast cancer cells was silenced, but present in normal breast tissues. EDNRB mRNA was expressed in BT549, MB231, T47D and SK-BR-3 cells. The expression level of EDN3 mRNA in breast cancer tissues was lower than that in the corresponding para-cancerous tissues （P = 0.000 6）. The recombinant lentivirus vector expressing EDN3 gene was successfully constructed and infected into BT549 and MB231 cells, pCDH-CMV-MCS-EF1-Puro-EDN3 had no influences on the proliferation of BT549 and MB231 cells （both P 〉 0.05）, but it could inhibit the migration and invasion of the cells （both P 〈 0.01）. Conclusion： The expression of EDN3 gene is downregulated in breast cancer tissues. EDN3 gene can inhibit the migration and invasion of breast cancer cells.