摘要
目的检测鼻咽癌细胞系中miR-143表达情况,建立稳定高表达miR-143的鼻咽癌细胞系,为研究其在鼻咽癌的发生和发展中的作用机制提供可靠的细胞模型。方法采用QPCR方法检测鼻咽癌细胞系CEN1、CNE2、HONE1、5-8F、6-10B及人永生化鼻咽上皮细胞NP69中miR-143的表达水平。构建重组逆转录病毒质粒pMSCV-puro-miR-143,包装逆转录病毒pMSCV-miR-143和pMSCV-Vector并感染鼻咽癌细胞,建立稳定高表达miR-143的鼻咽癌细胞系,并应用细胞黏附实验检测过表达mir-143后,鼻咽癌细胞对胞外基质的黏附性。结果鼻咽癌细胞系中miR-143的表达水平均低于对照组NP69细胞,其中高转移潜能细胞株5-8F的表达量最低,仅为对照组的3.03%,成瘤但不转移细胞株6-10B表达量最高,为对照组的63.59%。实时荧光定量PCR显示筛选后第7代细胞miR-143的表达水平比对照组高出达1080倍(P<0.05)。细胞黏附实验结果显示,过表达mir-143可降低5-8F细胞对胞外基质的黏附性。结论 miR-143在鼻咽癌细胞系中表达失常,其作用可能与鼻咽癌细胞的侵袭和转移相关;成功建立了稳定高表达miR-143的鼻咽癌细胞系,并初步确定miR-143能抑制5-8F细胞的黏附能力,为探索miR-143在鼻咽癌的发生和发展中的作用机制奠定了实验基础。
Objective To detect miR-143 expression in human nasopharyngeal carcinnoma（NPC） cell lines and explore the role of miR-143 in regulating the adhesion ability of NPC cells.Methods Fluorescence quantitative RT-PCR was used to detect miR-143 expression levels in 5 NPC cell lines（CEN1,CNE2,HONE1,5-8F,and 6-10B） and an immortalized human nasopharyngeal epithelial cell line（NP69）.The retrovirus plasmid pMSCV-puro-miR-143 was constructed and the packaged retroviruses pMSCV-puro and pMSCV-puro-miR-143 were infected in 5-8F cells to establish a cell line with stable miR-143 overexpression,whose adhesion ability was tested with adhesion assay.Results The expression of miR-143 was downregulated in the NPC cell lines,and the highly metastatic NPC cell line 5-8F had a expression of only 3.0% of the control level, as compared with the level of 63.59% in the tumorigenic but nonmetastatic NPC cell line 6-10B.The transfected 5-8F cells showed a 1080-fold increase of miR-143 expression（P0.05） with a significantly lowered adhesion ability.Conclusion miR-143 plays a role in regulating the invasiveness and metastasis of NPC,and overexpression of miR-143 causes a significant reduction of the adhesion ability of the highly metastatic NPC cell line 5-8F.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2013年第4期582-585,共4页
Journal of Southern Medical University
基金
广东省自然科学基金(S2011040003476)
