含WW域的氧化还原酶基因对肺癌转移的抑制作用

WW domain containing oxidoreductase gene inhibiting metastasis of lung cancer

目的观察含WW域的氧化还原酶基因（WWOX）基因对肺癌转移有无抑制作用。方法采用实时荧光定量聚合酶链反应（FQ—PCR）及Western blot检测手术后肺癌组织，高侵袭性肺癌细胞株A549、NCI—H1299及低侵袭性肺癌细胞株HBFA—E6／E7、NL9980的WWOX基因的表达。比较癌组织与肺癌旁组织、肺癌Ⅰ期与Ⅱ期＋Ⅲ期、高侵袭性肺癌细胞株与低侵袭性肺癌细胞株WWOX表达情况。将外源性WWOX基因稳定转染入WWOX表达阴性但高侵袭性的NCI—H1299细胞株，在WWOX表达阳性但低侵袭性的NL9980细胞株，应用小干扰RNA（siRNA）沉默WWOX的表达。应用Transwell法检测肺癌细胞的侵袭性。观察肺癌细胞株侵袭力的改变。结果WWOXmRNA在肺癌组织中的表达（0．03±0．01）明显低于癌旁组织（P〈0．05），在Ⅱ期＋Ⅲ期肺癌组织中的表达（0．24±0．09）明显低于I期肺癌组织（P〈0．05）。WWOXmRNA在高侵袭性H1299细胞中几乎不表达（0．91±0．01），而在低侵袭性NL9980细胞中呈现高表达（5．22±0．02）。Transwell小室显示转染了WWOX过表达质粒的H1299细胞数[（1156±133）个]与阴性对照细胞数[（3487±822）个]比较差异有统计学意义（P〈0．05），转染了WWOXsiRNA的NL9980细胞数[（1860±389）个]与阴性对照细胞数[（667±85）个]比较差异有统计学意义（P〈0．05）。且在H1299＋WWOX细胞中基质金属蛋白酶（MMP）-2（0．58±0．06）及波形蛋白（Vimentin，0．24±0．09）比H1299＋Mock表达均减少，而NL9980＋WWOXsiRNA细胞中MMP-2（1．51±0．08）及Vimentin（2．08±0．04）比NL9980＋NCsiRNA表达有增加。结论WWOX基因与肺癌转移相关，通过调控WWOX基因的表达可以对肺癌细胞的侵袭性产生明显的影响。

Objective To explore the inhibitory role of WW domain containing oxidoreductase （WWOX） gene during lung cancer cell metastases. Methods WWOX mRNA and protein levels were detected by real - time fluorescent quantitative polymerase chain reaction （ FQ - PCR） analysis and Western blotting lung cancer tissues, and lung cell lines. We induced WWOX over - expression by transient transfection of WWOX plasmid in highly invasive lung cancer cell NCI - H1299, and inhibited WWOX expression by transient transfection of WWOX small interfering RNA （siRNA） in poorly invasive lung cancer cell NL9980. The invasive properties of H1299 and NL9980 transfectants were compared using a Transwell chamber assay. Results The WWOX mRNA expression was significantly lower in lung carcinomas （0. 03 ±0. 01 ） than in normal tissues, and that was also lower in stage Ⅱ/Ⅲ lung carcinomas （0. 24 ± 0. 09） than in stage I tumors. The WWOX expression was lower in NCI - H1299 cells than in NL9980 ceils. The number of NCI - H1299 cells with WWOX over - expression （ 1 156 ± 133 ） were less than the mock transfectants （3 487 ±822 ）, the number of WWOX siRNA NL9980 transfectants （ 1 860 ±389 ） were more than the negetive control siRNA NL9980 transfectants （667± 85）. In addition, NCI -H1299 ＋WWOX cells expressed less matrix metalloproteinase （MMP） - 2 （ 0. 58 ± 0.06 ） and vimentin （ 0. 24 ± 0. 09） , and NL9980 cells transfected with WWOX siRNA expressed more MMP -2 （ 1.51 ±0.08） and vimentin （2. 08 ± 0. 04） than their respective controls. Conclusion WWOX gene is associated with metastases of lung cancer. WWOX can affect the invasive capacities of lung cells by regulating 13 - catenin signa- ling pathway target molecules such as vimentin and MMP -2.