miR-1254靶向GLTSCR2调控非小细胞肺癌增殖和活力
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摘要
目的探讨miR-1254、GLTSCR2与非小细胞肺癌凋亡和增殖活力的关系。方法检测非小细胞肺癌和正常肺细胞中miR-1254与GLTSCR2的表达;运用TargetScan在线分析miR-1254与GLTSCR2的相关性;利用luciferase assay检测miR-1254是否靶向调控GLTSCR2;转染miR-1254 mimics或miR-1254 inhibitor进非小细胞肺癌A549细胞,通过Western blot检测GLTSCR2的表达量和非小细胞肺癌A549细胞凋亡标志蛋白表达情况;通过CCK-8试验检测不同条件下A549细胞的增殖活力。结果非小细胞肺癌A549的miR-1254相对表达量最高(P <0.05); GLTSCR2在非小细胞肺癌A549和H1299表达量最低(P<0.05);miR-1254靶向GLTSCR2的3'UTR的193-200区域;miR-1254过表达时,GLTSCR2的表达量下降(P<0.05),细胞凋亡相关蛋白BAK的表达量明显下降(P<0.05),而BCL-2的表达量明显上升(P<0.05),并且CASPASE9/CLEAVED-CAS9的比例明显上升(P<0.05),细胞增殖活力明显上升(P<0.05);敲低miR-1254后,GLTSCR2的表达量明显上升(P<0.05),细胞凋亡相关蛋白BAK的表达量明显上升(P<0.05),而BCL-2的表达量明显下降(P <0. 05),并且CASPASE9/CLEAVED-CAS9的比例明显下降(P <0.05),细胞增殖活力明显下降(P<0.05)。结论 miR-1254能通过降低GLTSCR2的表达来降低非小细胞肺癌细胞凋亡以及提高非小细胞肺癌A549的增殖和活力。
Objective To investigate the relationship between miR-1254, GLTSCR2 and apoptosis and proliferation of non-small cell lung cancer. Methods We measured miR-1254 and GLTSCR2 expressions in non-small cell lung cancer and normal lung cells. TargetScan was used to analyze the correlation between miR-1254 and GLTSCR2. The luciferase assay was used to detect whether miR-1254 targeted GLTSCR2.Transfection of miR-1254 mimics or miR-1254 into non-small cell lung cancer A549 cells was performed and then tested by Western blot. The expression of GLTSCR2 and the expression of apoptosis marker protein in non-small cell lung cancer A549 cells were detected. The proliferation activities of A549 cells under different conditions were detected by CCK-8 assay. Results The relative expression of miR-1254 in non-small cell lung cancer A549 was the highest(P <0.05); GLTSCR2 was the lowest in non-small cell lung cancer A549 and H1299(P <0.05); miR-1254 targeted the 193-200 region of the 3'UTR of GLTSCR2; when overexpressing miR-1254,the expression of GLTSCR2 was decreased(P < 0. 05), and the expression of apoptosis-related protein BAK was significantly decreased(P <0. 05), while the expression of BCL-2 increased significantly(P <0.05) and the proportion of CASSASE9/CLEAVED-CAS9 increased significantly(P <0. 05),also the cell proliferation activity increased significantly(P <0. 05); We then knocked down miR-1254 after the overexpression,the expression of GLTSCR2 was significantly increased(P <0. 05), the expression of apoptosis-related protein BAK was significantly increased(P <0. 05),and the expression of BCL-2 was significantly decreased(P <0. 05), and the proportion of CASSASE9/CLEAVED-CAS9 decreased significantly(P < 0. 05), also the cell proliferation activity decreased significantly(P < 0. 05).Conclusion miR-1254 can reduce the apoptosis of non-cell lung cancer cells and promote the proliferation and viability of non-small cell lung cancer A549 by decreasing the expression of GLTSCR2.
Objective To investigate the relationship between miR-1254, GLTSCR2 and apoptosis and proliferation of non-small cell lung cancer. Methods We measured miR-1254 and GLTSCR2 expressions in non-small cell lung cancer and normal lung cells. TargetScan was used to analyze the correlation between miR-1254 and GLTSCR2. The luciferase assay was used to detect whether miR-1254 targeted GLTSCR2.Transfection of miR-1254 mimics or miR-1254 into non-small cell lung cancer A549 cells was performed and then tested by Western blot. The expression of GLTSCR2 and the expression of apoptosis marker protein in non-small cell lung cancer A549 cells were detected. The proliferation activities of A549 cells under different conditions were detected by CCK-8 assay. Results The relative expression of miR-1254 in non-small cell lung cancer A549 was the highest(P <0.05); GLTSCR2 was the lowest in non-small cell lung cancer A549 and H1299(P <0.05); miR-1254 targeted the 193-200 region of the 3'UTR of GLTSCR2; when overexpressing miR-1254,the expression of GLTSCR2 was decreased(P < 0. 05), and the expression of apoptosis-related protein BAK was significantly decreased(P <0. 05), while the expression of BCL-2 increased significantly(P <0.05) and the proportion of CASSASE9/CLEAVED-CAS9 increased significantly(P <0. 05),also the cell proliferation activity increased significantly(P <0. 05); We then knocked down miR-1254 after the overexpression,the expression of GLTSCR2 was significantly increased(P <0. 05), the expression of apoptosis-related protein BAK was significantly increased(P <0. 05),and the expression of BCL-2 was significantly decreased(P <0. 05), and the proportion of CASSASE9/CLEAVED-CAS9 decreased significantly(P < 0. 05), also the cell proliferation activity decreased significantly(P < 0. 05).Conclusion miR-1254 can reduce the apoptosis of non-cell lung cancer cells and promote the proliferation and viability of non-small cell lung cancer A549 by decreasing the expression of GLTSCR2.
引文
[1]丁碧莎,孙月娟,王海燕.DC-CIK联合化疗治疗非小细胞肺癌的研究进展[J].中国免疫学杂志,2016,32(6):916-919.
[2]蔡忠,李婷,陈烨,等.靶向EGFR分子的荧光探针设计及其对非小细胞肺癌体内外检测功能的研究[J].标记免疫分析与临床,2018,25(1):39-43.
[3]段桦.227例Ⅲb/Ⅳ期非小细胞肺癌患者生存期回顾性研究[D].北京中医药大学,2018.
[4] YOON J C, LING A J, ISIK M, et al. GLTSCR2/PICT1 links mitochondrial stress and Myc signaling[J]. Proc Natl Acad Sci USA,2014,111(10):3781-3186.
[5] PORSCH R M,MERELLO E,DE M P,et al. Sacral agenesis:a pilot whole exome sequencing and copy number study[J]. BMC Med Genet,2016,17(1):98.
[6] DU M,ZHANG Y,MAO Y,et al. MiR-33a suppresses proliferation of NSCLC cells via targeting METTL3 mRNA[J]. Biochem Biophys Res Commun,2016,482(4):582-589.
[7] LIU F,SONG D,WU Y, et al. MiR-155 inhibits proliferation and invasion by directly targeting PDCD4 in non-small cell lung cancer[J]. Thoracic Cancer,2017,8(6):613-619.
[8] WANG M, MENG B, LIU Y, et al. MiR-124 inhibits growth and enhances radiation-induced apoptosis in non-small cell lung cancer by inhibiting STAT3[J]. Cell Physiol Biochem,2017,44(5):2017-2028.
[9]彭华,王佳,赵玫,等.非小细胞肺癌血清microRNAs诊断模型的建立[C].中国肿瘤内科进展中国肿瘤医师教育,2014:727.
[10]刘悦超,邢影,蔡莉.Hippo信号通路在肺癌中的研究进展[J].中国肺癌杂志,2017,20(9):629-634.
[11]雷震霄.中医药治疗肺癌的研究进展[J].湖南中医杂志,2017,33(12):154-156.
[12]檀叶青,邓书婧,陈文儒.EGFR突变在非小细胞肺癌患者治疗应用中的研究进展[J].标记免疫分析与临床,2017,24(10):1197-1200.
[13]张一帆.RPS15A在非小细胞肺癌谷氨酰胺代谢途径中的调控作用[D].吉林大学,2016.
[14] ZHAN M,WEN F,LIU L, et al. JMJD1A promotes tumorigenesis and forms a feedback loop with EZH2/let-7c in NSCLC cells[J].Tumor Biol,2016,37(8):11237-11247.
[15] CHEN L,KONG G,ZHANG C,et al. MicroRNA-432 functions as a tumor suppressor gene through targeting E2F3 and AXL in lung adenocarcinoma[J]. Oncotarget,2016,7(15):20041-20053.
[16] CHU Y M,PENG H X,XU Y,et al. MicroRNA-1254 inhibits the migration of colon adenocarcinoma cells by targeting PSMD10[J]. J Dig Dis,2017,18(3):169-178.
[17] LU M,CHEN W H, WANG C Y,et al. Reciprocal regulation of miR-1254 and c-Myc in oral squamous cell carcinoma suppresses EMT-mediated metastasis and tumor-initiating properties through MAPK signaling[J]. Biochem Biophys Res Commun, 2017,484(4):801-807.
[18] LU M,CHEN W H, WANG C Y, et al. Retraction notice to”Reciprocal regulation of miR-1254 and c-Myc in oral squamous cell carcinoma suppresses EMT-mediated metastasis and tumor-initiating properties through MAPK signaling[Biochem. Biophys. Res.Commun. 484(4)(2017)801-807].[J]. Biochem Biophys Res Commun,2018,497(4):1186.
[19] WANG P, MENG W, HAN S C, et al. The nucleolar protein GLTSCR2 is required for efficient viral replication[J]. Sci Rep,2016,6:36226.
[20] FOUZ N,AMID A, HASHIM H Y. Gene expression analysis in MCF-7 breast cancer cells treated with recombinant bromelain[J].Appl Biochem Biotechnol,2014,173(7):1618-1639.
[21] LEE S, KIM J Y, KIM Y J, et al. Nucleolar protein GLTSCR2stabilizes p53 in response to ribosomal stresses[J]. Cell Death Differ,2012,19(10):1613-1622.
[22]姜莉,李玉榕.基于连续-离散扩展卡尔曼建模的p53-Mdm2调控关系研究[J].中国生物医学工程学报,2017,36(2):180-186.
[23] ROSE C M,ISASA M, ORDUREAU A, et al. Highly multiplexed quantitative mass spectrometry analysis of ubiquitylomes[J]. Cell Syst,2016,3(4):395-403.
[2]蔡忠,李婷,陈烨,等.靶向EGFR分子的荧光探针设计及其对非小细胞肺癌体内外检测功能的研究[J].标记免疫分析与临床,2018,25(1):39-43.
[3]段桦.227例Ⅲb/Ⅳ期非小细胞肺癌患者生存期回顾性研究[D].北京中医药大学,2018.
[4] YOON J C, LING A J, ISIK M, et al. GLTSCR2/PICT1 links mitochondrial stress and Myc signaling[J]. Proc Natl Acad Sci USA,2014,111(10):3781-3186.
[5] PORSCH R M,MERELLO E,DE M P,et al. Sacral agenesis:a pilot whole exome sequencing and copy number study[J]. BMC Med Genet,2016,17(1):98.
[6] DU M,ZHANG Y,MAO Y,et al. MiR-33a suppresses proliferation of NSCLC cells via targeting METTL3 mRNA[J]. Biochem Biophys Res Commun,2016,482(4):582-589.
[7] LIU F,SONG D,WU Y, et al. MiR-155 inhibits proliferation and invasion by directly targeting PDCD4 in non-small cell lung cancer[J]. Thoracic Cancer,2017,8(6):613-619.
[8] WANG M, MENG B, LIU Y, et al. MiR-124 inhibits growth and enhances radiation-induced apoptosis in non-small cell lung cancer by inhibiting STAT3[J]. Cell Physiol Biochem,2017,44(5):2017-2028.
[9]彭华,王佳,赵玫,等.非小细胞肺癌血清microRNAs诊断模型的建立[C].中国肿瘤内科进展中国肿瘤医师教育,2014:727.
[10]刘悦超,邢影,蔡莉.Hippo信号通路在肺癌中的研究进展[J].中国肺癌杂志,2017,20(9):629-634.
[11]雷震霄.中医药治疗肺癌的研究进展[J].湖南中医杂志,2017,33(12):154-156.
[12]檀叶青,邓书婧,陈文儒.EGFR突变在非小细胞肺癌患者治疗应用中的研究进展[J].标记免疫分析与临床,2017,24(10):1197-1200.
[13]张一帆.RPS15A在非小细胞肺癌谷氨酰胺代谢途径中的调控作用[D].吉林大学,2016.
[14] ZHAN M,WEN F,LIU L, et al. JMJD1A promotes tumorigenesis and forms a feedback loop with EZH2/let-7c in NSCLC cells[J].Tumor Biol,2016,37(8):11237-11247.
[15] CHEN L,KONG G,ZHANG C,et al. MicroRNA-432 functions as a tumor suppressor gene through targeting E2F3 and AXL in lung adenocarcinoma[J]. Oncotarget,2016,7(15):20041-20053.
[16] CHU Y M,PENG H X,XU Y,et al. MicroRNA-1254 inhibits the migration of colon adenocarcinoma cells by targeting PSMD10[J]. J Dig Dis,2017,18(3):169-178.
[17] LU M,CHEN W H, WANG C Y,et al. Reciprocal regulation of miR-1254 and c-Myc in oral squamous cell carcinoma suppresses EMT-mediated metastasis and tumor-initiating properties through MAPK signaling[J]. Biochem Biophys Res Commun, 2017,484(4):801-807.
[18] LU M,CHEN W H, WANG C Y, et al. Retraction notice to”Reciprocal regulation of miR-1254 and c-Myc in oral squamous cell carcinoma suppresses EMT-mediated metastasis and tumor-initiating properties through MAPK signaling[Biochem. Biophys. Res.Commun. 484(4)(2017)801-807].[J]. Biochem Biophys Res Commun,2018,497(4):1186.
[19] WANG P, MENG W, HAN S C, et al. The nucleolar protein GLTSCR2 is required for efficient viral replication[J]. Sci Rep,2016,6:36226.
[20] FOUZ N,AMID A, HASHIM H Y. Gene expression analysis in MCF-7 breast cancer cells treated with recombinant bromelain[J].Appl Biochem Biotechnol,2014,173(7):1618-1639.
[21] LEE S, KIM J Y, KIM Y J, et al. Nucleolar protein GLTSCR2stabilizes p53 in response to ribosomal stresses[J]. Cell Death Differ,2012,19(10):1613-1622.
[22]姜莉,李玉榕.基于连续-离散扩展卡尔曼建模的p53-Mdm2调控关系研究[J].中国生物医学工程学报,2017,36(2):180-186.
[23] ROSE C M,ISASA M, ORDUREAU A, et al. Highly multiplexed quantitative mass spectrometry analysis of ubiquitylomes[J]. Cell Syst,2016,3(4):395-403.
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