SPRY4-IT1通过激活锌指蛋白703基因表达增强肺癌细胞增殖能力  

SPRY4 - IT1 contributes to increased cell proliferation by activating zinc finger protein 703 expres- sion in non - small cell lung cancer

作　　者：胡宝利[1] 王作培[1] 张锋[1] 张海峰[1] 王晓龙[1] 韦海涛[1] 周勇安[2] 

机构地区：[1]河南大学淮河医院胸心外科,开封475000 [2]第四军医大学唐都医院胸外科,西安710032

出　　处：《中华实验外科杂志》2017年第2期249-251,共3页Chinese Journal of Experimental Surgery

基　　金：河南省教育厅科学技术研究重点项目（16A320034）

摘　　要：目的观察长链非编码RNA SPRY4-IT1在非小细胞肺癌（NSCLC）患者中的表达及其调控细胞增殖的分子生物学功能。 方法采用实时定量反转录聚合酶链反应（RT-qPCR）检测32例NSCLC患者癌组织和远癌正常肺组织中SPRY4-IT1的相对表达量；肺癌A549细胞株内转染针对SPRY4-IT1序列的小干扰RNA（si-SPRY4-IT1）和无关序列（si-nc），采用细胞计数试剂盒（CCK-8）实验计数两组细胞增殖情况并绘制生长曲线；采用结晶紫染色实验观察细胞克隆形成情况。采用Western blot检测转染si-SPRY4-IT1后A549细胞中锌指蛋白703（ZNF703）蛋白表达。 结果SPRY4-IT1在NSCLC患者癌组织和癌旁组织中的相对表达量分别为3.24±2.01和1.12±0.78，癌组织中的相对表达量显著高于癌旁组织（t＝5.560，P＝0.001）；非小细胞肺癌细胞株A549转染si-SPRY4-IT1后，细胞中SPRY4-IT1表达水平显著降低（P＝0.001）；CCK-8实验显示，转染si-SPRY4-IT1的A549细胞增殖能力显著降低（P＝0.001）；结晶紫染色显示，转染si-SPRY4-IT1的A549细胞克隆形成能力显著降低（P＝0.001）；转染si-SPRY4-IT1和si-nc的A549肺癌细胞中ZNF703 mRNA相对表达量分别为0.61±0.07和0.96±0.08，si-SPRY4-IT1细胞显著低于si-nc细胞（P＝0.001）；ZNF703蛋白相对表达量分别为0.08±0.02和0.12±0.03，si-SPRY4-IT1细胞显著低于si-nc细胞（P＝0.001）。 结论长链非编码RNA SPRY4-IT1在NSCLC中高表达，并可通过增强ZNF703表达促进肺癌细胞的增殖。Objective To observe the long non coding RNA SPRY4 -IT1 expression level in non -small cell lung cancer and its biological funciton in aspects of cell proliferation. Methods Real - time quantitative reverse transcriptase - polymerase chain reaction （ RT - qPCR） was used to measure the expression of SPRY4 - IT1 in 32 non - small cell lung cancer （NSCLC） patients. Loss of function ap- proach was then applied to confirm the biological function, especially cell prolifieration in cultured human lung cancer cells A5549, by cell counting kit - 8 （ CCK - 8） and clonogenie assay. We further used west- ern blot to reveal the activation of zinc finger protein 703 （ ZNF703 ） by SPRY4 - IT1. Results The SPRY4 - IT1 relative expression level were 1.12 ± 0. 78 and 3.24 ± 2. 01 respectively for tumor tissue and norma tissue of NSCLC patients with statistical difference （ t = 5. 56, P = 0. 001 ） ; SPRY4 - IT1 expression level were significant decreased by transinfection of si - SPRY4 - IT1 in A549 cells. A549 cell clone forma- tion ability was significantly reduced by traninfection of si - SPRY4 - IT1. The mRNA expression of ZNF＇/03 were 0. 61± 0. 07 and 0. 96 ± 0. 08 for si - SPRY4 - IT1 and si - nc A549 cells with statistical difference （ P = 0. 001 ） ; The protein expression of ZNFT03 were 0. 08 ± 0. 02 and 0. 12 ± 0. 03 for si - SPRY4 - IT1 and si - nc A549 cells with statistical difference （P =0. 001 ）. Conclusion SPRY4 - IT1 expression is up - regulated in tumor tissue of non - small cell lung cancer patients and the SPRY4 - IT1/ZNFT03 axis might contribute to increased proliferation ability of lung cancer ceils.

关 键 词：SPRY4-IT1 非小细胞肺癌 长链非编码RNA 增殖能力 

分 类 号：R734.2[医药卫生—肿瘤]