miR-424*在X射线照射后的A549细胞体内、外及非小细胞肺癌组织和血清中的表达
The expression of miR-424* in vivo and in vitro irradiated A549 cells, tissue and serum samples of non-small cell lung cancer
目的 研究2、4 GyX射线照射后人肺腺癌细胞miR-424*体内、外表达以及非小细胞肺癌患者肺组织及血清中miR-424*的表达变化及其意义.方法 2、4 GyX射线分别照射体外培养的A549细胞,以实时定量PCR(RT-qPCR)法检测A549细胞中miR-424*表达水平;用照射后的A549细胞制备裸鼠肺转移动物模型,检测裸鼠肺组织及血清中miR-424*的表达水平;收集肺癌患者肺组织及血清样本,检测miR424*表达水平.结果 2、4 GyX射线照射后1、2、12、24及48 h,miR-424*表达均显著升高(2 Gy:t=-45.886 ~-6.709,P<0.05;4 Gy:t=-29.087~-7.833,P<0.05);0、2、4 Gy照射后,miR-424*在裸鼠肺及血清中表达水平分别为空白对照组的9.72、8.58及4.7与11.93、9.22及8.99倍(t=-13.243 ~-3.052,P<0.05).6/11例(54.5%)患者肺癌组织中高表达miR-424*,腺癌、鳞癌病理类型间检出率差异无统计学意义(P>0.05);43/84例(51.20%)肺癌患者与健康志愿者相比,血清miR-424*表达升高1.97 ~ 17.71倍,其中腺癌患者血清检出率为39.1% (18/46),鳞癌患者血清检出率为65.8% (25/38),两种病理类型检出率差异有统计学意义(t =5.919,P<0.05);此外,84例肺癌患者中,miR-424*在未接受放疗的肺癌患者血清中的阳性检出率为41.5% (22/53),显著低于接受放疗的肺癌患者血清的阳性检出率67.7% (21/31)(t=5.387,P<0.05).结论 2、4GyX射线照射可增加A549细胞miRNA-424*的体内、外表达水平,可能与增强A549细胞体内、外侵袭转移能力有关.肺癌患者中50%以上的肺癌组织及肺癌患者血清中miR424*表达水平显著升高,可能与肺癌的病理类型及放疗相关.
更多Objective To investigate the expression of miR-424* in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of non-small cell lung cancer(NSCLC) patients,and to explore its potential role in the diagnosis and prognosis of lung cancer.Methods A549 cells were irradiated with 2 and 4 Gy X-rays,and some of irradiated cells were injected into nude mice through tail vein.Real time quantitative PCR (RT-qPCR) assay was employed to detect the expression of miR-424 * in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of lung cancer patients.Results Compared with the control group,the expression of miR-424* was up-regulated significantly in X-ray irradiated A549 cells at 1,2,12,24 and 48 hpost irradiation,respectively (2 Gy:t =-45.886--6.709,P <0.05;4 Gy:t =-29.087--7.833,P < 0.05).Furthermore,the expression of miR-424 * was up-regulated in the lung and serum of nude mice with injection of 0,2 and 4 Gy X-ray irradiated A549 cells,compared with control group (fold change was 9.72,8.58 and 4.7 with 2 Gy irradiation and 11.93,9.22 and 8.99 with 4 Gy irradiation,t=-13.243,-12.409,-9.833 in lung andt=-6.436,-3.052,-3.609 in serum,respectively,P < 0.05).Out of 11 tissue samples of NSCLC patients,6 were detected with up-regulated miR-424* expression,and no significant discrepancy of miR-424* expression was detected in two type of NSCLC tissue samples.On the contrary,43 serum samples were detected with up-regulated miR-424* expression out of 84 serum samples (51.20%) of NSCLC patients (fold change range 1.97 to 17.71),and significant discrepancy of miR-424* expression was shown in two subtypes of NSCLC serum samples [adenocarcinoma:39.10% (18/46) and squamous carcinoma:65.8% (25/38)],as well as in serum samples of NSCLC patients with radiotherapy [41.5% (22/53)] and without radiotherapy [67.7% (21/31)] (t=5.919,5.387,P <0.05,respectively).Conclusions 2 and 4 Gy X-ray irradiation could up-regulate the expression of miR-424* in A549 cells,which might be correlated with the enhanced metastasis of A549 cells induced by X-ray in vivo and in vitro.Furthermore,the expression of miR-424* was up-regulated in over 50% of the tissue and serum samples of NSCLC patients,which might be correlated with the diagnosis of NSCLC subtype and prognosis of radiotherapy.
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