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目的 探讨长链非编码RNA母系表达基因3(MEG3)在肺癌组织中的表达及其调控肺癌细胞增殖、迁移的生物学功能.方法 应用实时定量聚合酶链反应(Real-time PCR)法检测48例非小细胞肺癌(NSCLC)患者癌组织和配对的远癌组织(距离癌组织超过5 cm)中MEG3的表达水平.在A549和H460肺癌细胞株中转染MEG3过表达质粒(Pc-DNA3.1-MEG3).分别利用噻唑蓝(MTT)实验和划痕实验分析MEG3过表达后对肺癌细胞增殖和迁移能力的影响.应用Western blot实验检测相关生物学功能分子标志物的表达水平.结果 与远癌正常肺组织比较,肺癌组织中MEG3表达下调者25例占52.08％,显著高于上调表达者(P＜0.05);A549和H460细胞中转染空载pcDNA3.1和pcDNA3.1-MEG3后,MEG3表达水平分别为0.48 ±0.04、0.62±0.05和10.36±1.78、11.66±2.01,差异均有统计学意义(P＜0.05);MTT实验显示,转染MEG3后A549和H460细胞的增殖能力显著下降(P＜0.05);划痕实验显示,转染MEG3后A549细胞的迁移能力显著降低(P＜0.05);Western blot分析细胞增殖相关蛋白细胞周期素D1 (Cyclin D1,CCND1)的表达水平,结果显示,MEG3过表达可显著抑制A549和H460肺癌细胞中CCND1表达(P＜0.05).结论 NSCLC中MEG3表达水平明显降低,MEG3可通过调控Cyclin D1的表达来影响细胞的增殖和迁移能力.

Objective To evaluate the maternally expressed gene 3 (MEG3) expression in non-small cellu luang cancer and its role proliferation and migration.Methods Fourty eight paties diagnosed of non-small cell lung cancer were included in this study.The MEG3 expression of the cancer tissue and normal lung tissue for the 48 cases were tested by real-time quantitative polymerase chain reaction (Real-time PCR) assay.The lung cancer cell line of A549 and H460 were transfected with long non-coding RNA MEG3 over expression plasmid (Pc-DNA3.1-MEG3).After transfection of Pc-DNA3.1-MEG3,the ablity of prolifieration and migration were tested by methyl thiazol tetrazolium (MTT) and wound assay.The associated protein fore prolifieration and migration were tested by Western blotting assay.Results Compared with normal lung tissue,25 (52.08％) cancer tissue were showed low expression of MEG3 with statistical difference (P ＜ 0.05);The expression of MEG3 were 0.48 ± 0.04,0.62 ± 0.05 and 10.36 ± 1.78,11.66 ±2.01 in A549 AND H460 lung cancer cell line for transfection of pcDNA3.1 and pcDNA3.1-MEG3 with statistical difference (P ＜ 0.05);The proliferation and migration ability was significant decreased after transfection of pcDNA3.1-MEG3 for A549 and H460 lung cancer cells (P ＜ 0.05).And the cyclin D1 protein expression was significant inhibit after transfection of pcDNA3.1-MEG3 (P ＜ 0.05).Conclusion MEG3 was relative low expressed in non-small cell lung cancer (NSCLC) and it inhibits proliferation and migration through inbibiting Cyclin D1 expression.