siRNA沉默WFDC2对肺癌A549细胞增殖和侵袭的影响
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摘要
目的初步探讨siRNA沉默WFDC2对A549肺癌细胞增殖、侵袭的影响及可能机制。方法构建WFDC2特异性小分子干扰RNA(siRNA),脂质体法转染肺癌细胞系A549;RT-PCR和Westernblot分别检测mRNA及蛋白表达量;CCK8实验检测细胞增殖速度;划痕实验及Transwell检测细胞侵袭、迁移能力。结果 siRNA-WFDC2转染组A549较空白组及阴性对照组mRNA表达量与蛋白表达量显著降低(P <0.05),细胞的增殖受到显著抑制(P <0.05),并随着时间的延长抑制明显,细胞迁移距离及细胞穿膜数均显著降低(P <0.05)。结论 siRNA沉默WFDC2基因的表达能够抑制肺癌A549细胞增殖、迁移及侵袭能力,WFDC2基因有可能成为肺癌治疗的新靶点。
Objective To investigate the influence of WFDC2 gene expression down-regulation by RNA interference(RN Ai) on proliferation and invasion of in human lung cancer A549 cells and mechanism. Methods Construction of WFDC2 specific Small interfering RNA(siRNA), siRNA-WFDC2 was transfected into lung cancer A549 cell lines by Lipofectamine 2000. The mRNA and protein expression of different groups about WFDC2 was detected by RT-PCR and Western blot assay. The CCK8 was used to detect the proliferation, and the scratch test and transwell were used to detect the ability of A549 cell lines on the migration and invasion. Results The results of RT-PCR and Western blot showed that the level of mRNA and protein in the transfection group were significantly lower than blank group and negative group. The proliferation rate of the A549 in siRN A group was inhibited(P < 0.05),and the inhibition was obvious with the prolongation of time. The migration distance and the number of perforated cells of the A549 in siRNA group was significantly decreased(P < 0.05). Conclusion Silencing the expression of WFDC2 gene by siRNA can inhibit the proliferation, migration, invasion of lung cancer A549 cell lines, WFDC2 gene may be a new target for lung cancer therapy.
Objective To investigate the influence of WFDC2 gene expression down-regulation by RNA interference(RN Ai) on proliferation and invasion of in human lung cancer A549 cells and mechanism. Methods Construction of WFDC2 specific Small interfering RNA(siRNA), siRNA-WFDC2 was transfected into lung cancer A549 cell lines by Lipofectamine 2000. The mRNA and protein expression of different groups about WFDC2 was detected by RT-PCR and Western blot assay. The CCK8 was used to detect the proliferation, and the scratch test and transwell were used to detect the ability of A549 cell lines on the migration and invasion. Results The results of RT-PCR and Western blot showed that the level of mRNA and protein in the transfection group were significantly lower than blank group and negative group. The proliferation rate of the A549 in siRN A group was inhibited(P < 0.05),and the inhibition was obvious with the prolongation of time. The migration distance and the number of perforated cells of the A549 in siRNA group was significantly decreased(P < 0.05). Conclusion Silencing the expression of WFDC2 gene by siRNA can inhibit the proliferation, migration, invasion of lung cancer A549 cell lines, WFDC2 gene may be a new target for lung cancer therapy.
引文
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[3]CHEN Y,MU X,WANG S,et al.WAP four-disulfide core domain protein 2 mediates the proliferation of human ovarian cancer cells through the regulation of growth and apoptosis associated genes[J].Oncol Rep,2013,29(1):288-296.
[4]朱亚飞,高国兰,褚芳.RNAi介导的WFDC2基因沉默抑制人卵巢浆液性上皮癌生长的体内实验[J].现代妇产科进展,2016,3(25):187-191.
[5]CHEN Y,WANG S H,LIU T C,et al.WAP four disculfide core domain protein 2 gene(WFDC2)is a target of estrogen in ovarian cancer cell[J].J ovarian res,2016,9(10):1-11.
[6]WOJCIK E,TARAPACZ J,RYCHLIK U,et al.Human epididymis protein 4(HE4)in patients with small-cell lung cancer[J].Clin Lab,2016,62(9):1625-1632.
[7]CHOI S I,JANG M A,JEON B R,et al.Clinical usefulness of human epididymis protein 4 in lung cancer[J].Ann lab Med,2017,37(6):526-530.
[8]WENHAI HUANG,SHOUYUN WU,ZHICHAO LIN,et al.Evaluation of HE4 in the diagnosis and follow up of non-small cell lung cancers[J].Clin Lab,2017,63(3):461-467.
[9]ZHONG H,QIAN Y,FANG S,et al.HE4 expression in lung cancer,a meta-analysis[J].Clin chim Acta,2017,470:109-114.
[10]WAGENBLAST E,SOTO M,GUTIéRREZ-áNGEL S,et al.Amodel of breast cancer heterogeneity reveals vascular mimicry as a driver of metastasis[J].Nature,2015,520(7547):358-362.
[11]LU Q,CHEN H,SENKOWSKI C,et al.Recombinant HE4protein promotes proliferation of pancreatic and endometrial cancer cell lines[J].Oncol Rep,2016,35(1):163-170.
[12]YILMAZ S A,ALTINKAYA S O,KERIMOGLU O S,et al.The role of human epididymis secretory protein E4 in patients with endometrial cancer and premalignant endometrial lesion[J].JObstet Gynaecol,2017,37(1):58-63.
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[14]RIBEIRO J R,SCHORL C,YANO N,et al.HE4 promotes collateral resistance to cisplatin and paclitaxel in ovarian cancer cells[J].J Ovarian Res,2016,9(28):1-18.
[15]MOORE R G,HILL E K,HORANT T,et al.HE4(WFDC2)gene overexpression promotes ovarian tumor growth[J].Sci Rep,2014,4:3574.doi:10.1038/srep03574.
[16]ZENG Q,LIU M,ZHOU N,et al.,Serum human epididymis protein 4(HE4)may be a better tumor marker in early lung cancer[J].Clin Chim Acta,2016,455:102-106.
[17]罗佳龄,彭霞,林堃,等.WFDC2编码的HE4在肺癌诊断和疗效监测中的应用价值[J].检验医学,2018,33(6):508-511.
[18]ZHU L,ZHUANG H,WANG H,et al.Overexpression of HE4(human epididymis protein 4)enhances proliferation,invasion and metastasis of ovarian cancer[J].Oncotarget,2016,7(1):729-744.