长链非编码RNA MAPKAPK5-AS1对肺癌细胞的侵袭和凋亡的影响
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摘要
目的研究长链非编码RNA(long ncRNA, lncRNA)MAPKAPK5-AS1对肺癌细胞的侵袭和凋亡的影响。方法 real-time PCR法筛选MAPKAPK5-AS1高表达和低表达肺癌细胞系,将MAPKAPK5-AS1过表达载体转染低表达细胞系,MAPKAPK5-AS1小干扰RNA (small interference RNA,siRNA)转染高表达细胞系,Western blot分别检测宿主基因MAPKAPK5和凋亡蛋白表达,transwell实验分析细胞侵袭能力。结果 MAPKAPK5-AS1在肺癌H2291细胞株和A549细胞株中高表达,而在H441细胞株中低表达。在H441细胞中过表达MAPKAPK5-AS1后, MAPKAPK5表达明显下降,侵袭能力明显增强(F=319.5,P<0.01),凋亡蛋白Cleaved caspase 9的表达下降;而在H2291细胞和A549细胞中分别干扰MAPKAPK5-AS1表达后,MAPKAPK5表达明显升高,侵袭能力明显下降(F值分别为417.1、530.2,P<0.01),凋亡蛋白Cleaved caspase 9的表达升高。结论 MAPKAPK5-AS1可能作为原癌基因通过调控宿主基因MAPKAPK5表达而增强肺癌细胞侵袭能力,同时抑制细胞凋亡。
Objective To investigate the effects of long-chain non-coding RNA(long ncRNA, lncRNA) MAPKAPK5-AS1 on invasion and apoptosis of lung cancer cells. Methods Real time PCR was performed to screen the lung cancer cells with high or low expression MAPKAPK5-AS1, MAPKAPK5-AS1 over expression vector or MAPKAPK5-AS1 small interference RNA(siRNA) was transfected into the screened lung cancer cells, respectively. The host gene MAPKAPK5 and apoptosis proteins were detected by western blot, the cellular invasive ability was analyzed by transwell assay. Results MAPKAPK5-AS1 expression was relatively higher in the H2291 and A549 cells than the H441 cells. After MAPKAPK5-AS1 over expression in the H441 cells, host gene MAPKAPK5 expression was inhibited, the cellular invasive ability was increased(F=319.5, P<0.01), and apoptosis protein cleaved caspase 9 expression was decreased. Conversely, after MAPKAPK5-AS1 siRNA tansfection in the H2291 and A549 cells, host gene MAPKAPK5 expression was increased, the cellular invasive ability was weakened(F=417.1, F=530.2, P<0.01), and apoptosis protein cleaved caspase 9 expression was increased. Conclu-sion MAPKAPK5-AS1 may act as an oncogene to promote cellular invasion and inhibit cellular apoptosis of lung cancer cells by down regulating host gene MAPKAPK5 expression.
Objective To investigate the effects of long-chain non-coding RNA(long ncRNA, lncRNA) MAPKAPK5-AS1 on invasion and apoptosis of lung cancer cells. Methods Real time PCR was performed to screen the lung cancer cells with high or low expression MAPKAPK5-AS1, MAPKAPK5-AS1 over expression vector or MAPKAPK5-AS1 small interference RNA(siRNA) was transfected into the screened lung cancer cells, respectively. The host gene MAPKAPK5 and apoptosis proteins were detected by western blot, the cellular invasive ability was analyzed by transwell assay. Results MAPKAPK5-AS1 expression was relatively higher in the H2291 and A549 cells than the H441 cells. After MAPKAPK5-AS1 over expression in the H441 cells, host gene MAPKAPK5 expression was inhibited, the cellular invasive ability was increased(F=319.5, P<0.01), and apoptosis protein cleaved caspase 9 expression was decreased. Conversely, after MAPKAPK5-AS1 siRNA tansfection in the H2291 and A549 cells, host gene MAPKAPK5 expression was increased, the cellular invasive ability was weakened(F=417.1, F=530.2, P<0.01), and apoptosis protein cleaved caspase 9 expression was increased. Conclu-sion MAPKAPK5-AS1 may act as an oncogene to promote cellular invasion and inhibit cellular apoptosis of lung cancer cells by down regulating host gene MAPKAPK5 expression.
引文
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[6] Zhang J,Fan D,Jian Z,et al.Cancer specific long noncoding RNAs show differential expression patterns and competing endogenous RNA potential in hepatocellular carcinoma[J].PLoS ONE,2015,10(10):e0141042.
[7] Wang W,Zhao Z,Yang F,et al.An immune-related lncRNA signature for patients with anaplastic gliomas[J].J Neurooncol,2018,136(2):263-271.
[8] 蔡静,朱方超,俞海波,等.甲基化调控的长链非编码RNA-GAS5对肝星状细胞增殖及活化的影响[J].中国卫生检验杂志,2016,26(24):3497-3500.
[9] 孙一奎,赵枫,王峰,等.血浆长链非编码RNA H19在原发性肝癌诊断中的价值[J].中国卫生检验杂志,2016,26(3):305-307,310.
[10] 林姜鑫,王伟伟,王莉,等.长链非编码RNA(linc02471)对食管鳞状细胞癌细胞增殖和侵袭的影响[J].中国卫生检验杂志,2018,28(5):516-519.
[11] 周武,季丽丽,张美娟,等.前列腺癌细胞中特异性标志物lncRNAPCA3的异常表达及其与PRUNE2 mRNA间的相互调控[J].中国卫生检验杂志,2017,27(11):1595-1597.
[12] 裴东旭,李永伟,刘心伟,等.长链非编码RNA用于非小细胞肺癌诊断的Meta分析[J].中国卫生检验杂志,2017,27(17):2548-2550,2554.
[13] Zhang J,Zhu N,Chen X.A novel long noncoding RNA LINC01133 is upregulated in lung squamous cell cancer and predicts survival[J].Tumour Biol,2015,36(10):7465-7471.
[14] Kanzaki R,Naito H,Kise K,et al.Gas6 derived from cancer-associated fibroblasts promotes migration of Axl-expressing lung cancer cells during chemotherapy[J].Sci Rep,2017,7(1):10613.
[15] Li S,Mei Z,Hu HB,et al.The lncRNA MALAT1 contributes to non-small cell lung cancer development via modulating miR-124/STAT3 axis[J].J Cell Physiol,2018,233(9):6679-6688.
[16] Zhou C,Ye L,Jiang C,et al.Long noncoding RNA HOTAIR,a hypoxia-inducible factor-1alpha activated driver of malignancy,enhances hypoxic cancer cell proliferation,migration,and invasion in non-small cell lung cancer[J].Tumour Biol,2015,36(12):9179-9188.