敲除GLRX3基因对鼻咽癌细胞生物学的影响

Biological effects of knocking down GLRX3 in nasopharyngeal carcinoma cells

目的建立稳定敲除GLRX3(glutaredoxin 3)基因的鼻咽癌细胞株,探索GLRX3基因在鼻咽癌细胞中的生物学功能。方法构建人GLRX3基因的shRNA逆转录病毒载体,将其转染入鼻咽癌细胞株HONE1中,筛选、建立稳定敲除GLRX3基因的HONE1细胞株。采用实时荧光定量PCR法验证筛选细胞中GLRX3基因的转录水平,通过克隆形成实验和划痕实验检测敲除GLRX3基因后鼻咽癌细胞克隆形成能力及迁移运动能力,评价敲除GLRX3基因后鼻咽癌细胞生物学行为的改变情况。结果成功构建的4个GLRX3 shRNA-pBINNS2质粒载体均可抑制GLRX3基因的转录表达。敲除GLRX3基因的HONE1细胞的克隆形成能力及迁移运动能力均降低。结论敲除GLRX3基因可抑制鼻咽癌细胞的恶性生物学行为,GLRX3可能是鼻咽癌候选的癌基因。

Objective To establish glutaredoxin 3 （GLRX3）-knocked down nasopharyngeal carcinoma （ NPC ） cell lines and explore the biological functions of the GLRX3 gene. Methods Retroviral vectors expressing shRNA against GLRX3 were constructed and transfected into HONE1 ceils, and stable cell lines showing knocked-down GLRX3 expression were selected. Reduced GLRX3 tran- scription in the cell lines was confirmed by real-time quantitative PCR.The cells were examined in colony formation and wound heal- ing assays to explore the influence of GLRX3 on these processes. Results Four retroviral shRNA vectors targeting GLRX3（shRNA- pBINNS2） were constructed and stably transfected into HONE1 cells.GLRX3 transcription was effectively suppressed in the stable cell lines.Knocking down GLRX3 inhibited clonogenicity and migration. Conclusions Knocking down GLRX3 inhibits the malignant behavior of NPC cells, suggesting that GLRX3 is a candidate oncogene in NPC.