HOTAIR lncRNA对垂体生长激素腺瘤侵袭及激素分泌的影响
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摘要
目的探讨长链非编码RNA(long non-coding RNA,lncRNA)中的HOX基因的反义基因间RNA(HOX transcript anti-sense intergenic RNA,HOTAIR)即HOTAIR lncRNA对垂体生长激素腺瘤侵袭和激素分泌的影响。方法采用qRT-PCR技术检测非侵袭性和侵袭性垂体生长激素腺瘤及正常垂体组织中的HOTAIR lncRNA,分析其差异表达。将对数生长期垂体瘤细胞株GH3随机分为对照组、阴性对照组和HOTAIR lncRNA siRNA组:对照组不作特殊处理,阴性对照组转染对照siRNA质粒,HOTAIR lncRNA siRNA组转染HOTAIR lncRNA siRNA质粒,培养72 h,采用ELISA检测细胞培养液中的生长激素,采用Transwell技术观测细胞侵袭力。结果非侵袭性和侵袭性垂体生长激素腺瘤及正常垂体前叶组织中的HOTAIR lncRNA相对表达量分别为1.60±0.25、3.27±0.57、1.00±0.00;非侵袭性和侵袭性垂体生长激素腺瘤组织中的HOTAIR lncRNA相对表达量与正常垂体组织相比,P均<0.05;侵袭性垂体生长激素腺瘤组织中的HOTAIR lncRNA相对表达量与非侵袭性生长激素腺瘤相比,P<0.01。阴性对照组、HOTAIR lncRNA siRNA组的HOTAIR lncRNA mRNA相对表达量分别为1.00±0.00、0.50±0.20,两组相比,P<0.01;阴性对照组、HOTAIR lncRNA siRNA组的生长激素表达量分别为(1.00±0.00)ng/m L、(0.64±0.15)ng/m L,两组相比,P<0.05;阴性对照组、HOTAIR lncRNA siRNA组的穿膜细胞数分别为(177±11)、(57±9)个/视野,两组相比,P<0.05。结论垂体生长激素腺瘤组织中HOTAIR lncRNA mRNA表达量升高,侵袭性垂体生长激素腺瘤组织中的HOTAIR lncRNA相对表达量高于非侵袭性生长激素腺瘤。HOTAIR lncRNA mRNA的高表达可能导致垂体生长激素腺瘤的侵袭行为和生长激素过量分泌。
Objective To investigate the effects of HOX transcript anti-sense intergenic RNA( HOTAIR) in the long non-coding RNA( lncRNA)( HOTAIR lncRNA) on the invasion and hormone secretion of pituitary growth hormone( GH)adenoma. Methods The expression of HOTAIR lncRNA in the normal pituitary,non-invasive and invasive pituitary GH adenomas was examined by qRT-PCR. The pituitary tumor cell line GH3 in the logarithmic growth phase was randomly divided into the control group,the negative control group,and the HOTAIR lncRNA siRNA group: the control group received no special treatment,the negative control group was transfected with a control siRNA plasmid,and the HOTAIR lncRNA siRNA group was transfected with the HOTAIR lncRNA siRNA plasmid; they were cultured for 72 hours,and we tested GH by using ELISA,and observed its invasive ability by Transwell assay. Results The relative expression levels of HOTAIR lncRNA in the non-invasive and invasive pituitary GH adenomas and normal anterior pituitary tissues were 1. 60 ± 0. 25,3. 27 ± 0. 57,and 1. 00 ± 0. 00. The relative expression level of HOTAIR lncRNA in non-invasive and invasive pituitary GH tissues was higher than that in normal pituitary tissues( P < 0. 05). The relative expression level of HOTAIR lncRNA in the invasive pituitary GH adenomas was higher than that in non-invasive ones( P < 0. 01). The relative expression levels of HOTAIR lncRNA mRNA in the negative control group and the HOTAIR lncRNA siRNA group were 1. 00 ± 0. 00 and0. 64 ± 0. 15( P < 0. 05). The relative expression levels of GH in the negative control group and the HOTAIR lncRNA siRNA group were( 1. 00 ± 0. 00) and( 0. 64 ± 0. 15) ng/m L( P < 0. 05). The cell invasion inhibition rates of the negative control group and the HOTAIR lncRNA siRNA group were( 177 ± 11) and( 57 ± 9)/field,respectively( P < 0. 05).Conclusions The expression level of HOTAIR lncRNA in the pituitary growth hormone adenoma tissues increases,and the relative expression level of HOTAIR lncRNA in the invasive pituitary GH adenoma is higher than that in the non-invasive GH adenoma. High expression of HOTAIR lncRNA mRNA may lead to invasive behavior of pituitary growth hormone adenomas and excessive secretion of growth hormone.
Objective To investigate the effects of HOX transcript anti-sense intergenic RNA( HOTAIR) in the long non-coding RNA( lncRNA)( HOTAIR lncRNA) on the invasion and hormone secretion of pituitary growth hormone( GH)adenoma. Methods The expression of HOTAIR lncRNA in the normal pituitary,non-invasive and invasive pituitary GH adenomas was examined by qRT-PCR. The pituitary tumor cell line GH3 in the logarithmic growth phase was randomly divided into the control group,the negative control group,and the HOTAIR lncRNA siRNA group: the control group received no special treatment,the negative control group was transfected with a control siRNA plasmid,and the HOTAIR lncRNA siRNA group was transfected with the HOTAIR lncRNA siRNA plasmid; they were cultured for 72 hours,and we tested GH by using ELISA,and observed its invasive ability by Transwell assay. Results The relative expression levels of HOTAIR lncRNA in the non-invasive and invasive pituitary GH adenomas and normal anterior pituitary tissues were 1. 60 ± 0. 25,3. 27 ± 0. 57,and 1. 00 ± 0. 00. The relative expression level of HOTAIR lncRNA in non-invasive and invasive pituitary GH tissues was higher than that in normal pituitary tissues( P < 0. 05). The relative expression level of HOTAIR lncRNA in the invasive pituitary GH adenomas was higher than that in non-invasive ones( P < 0. 01). The relative expression levels of HOTAIR lncRNA mRNA in the negative control group and the HOTAIR lncRNA siRNA group were 1. 00 ± 0. 00 and0. 64 ± 0. 15( P < 0. 05). The relative expression levels of GH in the negative control group and the HOTAIR lncRNA siRNA group were( 1. 00 ± 0. 00) and( 0. 64 ± 0. 15) ng/m L( P < 0. 05). The cell invasion inhibition rates of the negative control group and the HOTAIR lncRNA siRNA group were( 177 ± 11) and( 57 ± 9)/field,respectively( P < 0. 05).Conclusions The expression level of HOTAIR lncRNA in the pituitary growth hormone adenoma tissues increases,and the relative expression level of HOTAIR lncRNA in the invasive pituitary GH adenoma is higher than that in the non-invasive GH adenoma. High expression of HOTAIR lncRNA mRNA may lead to invasive behavior of pituitary growth hormone adenomas and excessive secretion of growth hormone.
引文
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[14]Gupta RA,Shah N,Wang KC,et al.Long non-coding RNA HOTAIR reprograms chromatin state to promote cancer metastasis[J].Nature,2010,464(7291):1071-1076.
[15]Li D,Feng J,Wu T,et al.Long intergenic noncoding RNA HOTAIR is overexpressed and regulates pten methylation in laryngeal squamous cell carcinoma[J].Am J Pathol,2013,182(1):64-70.
[16]Yang Z,Zhou L,Wu LM,et al.Overexpression of long non-coding RNA HOTAIR predicts tumor recurrence in hepatocellular carcinoma patients following liver transplantation[J].Ann Surg Oncol,2011,18(5):1243-1250.
[17]Kogo R,Shimamura T,Mimori K,et al.Long noncoding RNA HOTAIR regulates polycomb-dependent chromatin modification and is associated with poor prognosis in colorectal cancers[J].Cancer Res,2011,71(20):6320-6326.
[18]Geng YJ,Xie SL,Li Q,et al.Large intervening non-coding RNA HOTAIR is associated with hepatocellular carcinoma progression[J].J Int Med Res,2011,39(6):2119-2128.
[19]Nakagawa T,Endo H,Yokoyama M,et al.Large noncoding RNA HOTAIR enhances aggressive biological behavior and is associated with short disease-free survival in human non-small cell lung cancer[J].Biochem Biophys Res Commun,2013,436(2):319-324.
[20]Niinuma T,Suzuki H,Nojima M,et al.Upregulation of MiR-196a and HOTAIR drive malignant character in gastrointestinal stromal tumors[J].Cancer Res,2012,72(5):1126-1136.
[21]Kim K,Jutooru I,Chadalapaka G,et al.HOTAIR is a negative prognostic factor and exhibits pro-oncogenic activity in pancreatic cancer[J].Oncogene,2013,32(13):1616-1625.
[22]Shi YJ,Matson C,Lan F,et al.Regulation of LSD1 histone demethylase activity by its associated factors[J].Mol Cell,2005,19(6):857-864.
[23]Li Z,Li C,Liu C,et al.Expression of the long non-coding RNAs MEG3,HOTAIR,and MALAT-1 in non-functioning pituitary adenomas and their relationship to tumor behavior[J].Pituitary,2015,18(1):42-47.
[24]李九州,张玉奇,赵全成,等.抑制HOTAIR的表达促进替莫唑胺引起的垂体腺瘤细胞凋亡[J].滨州医学院学报,2016,39(3):165-168.
[2]Mete O,Lopes MB.Overview of the 2017 who classification of pituitary tumors[J].Endocr Pathol,2017,28(3):228-243.
[3]Saeger W,Honegger J,Theodoropoulou M,et al.Clinical impact of the current who classification of pituitary adenomas[J].Endocr Pathol,2016,27(2):104-114.
[4]Feelders RA,de Herder WW,Neggers SJ,et al.Pasireotide,a multi-somatostatin receptor ligand with potential efficacy for treatment of pituitary and neuroendocrine tumors[J].Drugs Today(Barc),2013,49(2):89-103.
[5]Lavrentaki A,Paluzzi A,Wass JA,et al.Epidemiology of acromegaly:review of population studies[J].Pituitary,2017,20(1):4-9.
[6]Petersenn S,Schopohl J,Barkan A,et al.Pasireotide(SOM230)demonstrates efficacy and safety in patients with acromegaly:a randomized,multicenter,phase ii trial[J].J Clin Endocrinol Metab,2010,95(6):2781-2789.
[7]Li J,Meng H,Bai Y,et al.Regulation of lncRNA and its role in cancer metastasis[J].Oncol Res,2016,23(5):205-217.
[8]Slaby O,Laga R,Sedlacek O.Therapeutic targeting of non-coding RNAs in cancer[J].Biochem J,2017,474(24):4219-4251.
[9]Haemmerle M,Gutschner T.Long non-coding RNAs in cancer and development:where do we go from here[J].Int J Mol Sci,2015,16(1):1395-1405.
[10]Bhan A,Mandal SS.LncRNA HOTAIR:a master regulator of chromatin dynamics and cancer[J].Biochim Biophys Acta,2015,1856(1):151-164.
[11]Yu X,Li Z.Long non-coding RNA HOTAIR:A novel oncogene(review)[J].Mol Med Rep,2015,12(4):5611-5618.
[12]Mestron A,Webb SM,Astorga R,et al.Epidemiology,clinical characteristics,outcome,morbidity and mortality in acromegaly based on the spanish acromegaly registry(registro espanol de acromegalia,rea)[J].Eur J Endocrinol,2004,151(4):439-446.
[13]白吉伟,李储忠,桂松柏,等.垂体生长激素腺瘤患者的临床特点及手术疗效初步分析[J].中华神经外科杂志,2015,31(7):653-657.
[14]Gupta RA,Shah N,Wang KC,et al.Long non-coding RNA HOTAIR reprograms chromatin state to promote cancer metastasis[J].Nature,2010,464(7291):1071-1076.
[15]Li D,Feng J,Wu T,et al.Long intergenic noncoding RNA HOTAIR is overexpressed and regulates pten methylation in laryngeal squamous cell carcinoma[J].Am J Pathol,2013,182(1):64-70.
[16]Yang Z,Zhou L,Wu LM,et al.Overexpression of long non-coding RNA HOTAIR predicts tumor recurrence in hepatocellular carcinoma patients following liver transplantation[J].Ann Surg Oncol,2011,18(5):1243-1250.
[17]Kogo R,Shimamura T,Mimori K,et al.Long noncoding RNA HOTAIR regulates polycomb-dependent chromatin modification and is associated with poor prognosis in colorectal cancers[J].Cancer Res,2011,71(20):6320-6326.
[18]Geng YJ,Xie SL,Li Q,et al.Large intervening non-coding RNA HOTAIR is associated with hepatocellular carcinoma progression[J].J Int Med Res,2011,39(6):2119-2128.
[19]Nakagawa T,Endo H,Yokoyama M,et al.Large noncoding RNA HOTAIR enhances aggressive biological behavior and is associated with short disease-free survival in human non-small cell lung cancer[J].Biochem Biophys Res Commun,2013,436(2):319-324.
[20]Niinuma T,Suzuki H,Nojima M,et al.Upregulation of MiR-196a and HOTAIR drive malignant character in gastrointestinal stromal tumors[J].Cancer Res,2012,72(5):1126-1136.
[21]Kim K,Jutooru I,Chadalapaka G,et al.HOTAIR is a negative prognostic factor and exhibits pro-oncogenic activity in pancreatic cancer[J].Oncogene,2013,32(13):1616-1625.
[22]Shi YJ,Matson C,Lan F,et al.Regulation of LSD1 histone demethylase activity by its associated factors[J].Mol Cell,2005,19(6):857-864.
[23]Li Z,Li C,Liu C,et al.Expression of the long non-coding RNAs MEG3,HOTAIR,and MALAT-1 in non-functioning pituitary adenomas and their relationship to tumor behavior[J].Pituitary,2015,18(1):42-47.
[24]李九州,张玉奇,赵全成,等.抑制HOTAIR的表达促进替莫唑胺引起的垂体腺瘤细胞凋亡[J].滨州医学院学报,2016,39(3):165-168.