长链非编码RNA HOST2对胰腺癌细胞增殖迁移和侵袭的影响
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摘要
目的:探讨长链非编码RNA(lncRNA)HOST2对胰腺癌细胞增殖、迁移和侵袭的影响及机制。方法:qRT-PCR检测正常胰腺上皮细胞系HPDE6-C7及胰腺癌细胞系Panc-1、AsPC-1、BxPC-3、HPAC中lncRNAHOST2表达水平。将Panc-1细胞分别转染siRNA-HOST2(si-HOST2组)和阴性对照序列(阴性对照组)后,用MTT法测定细胞增殖,细胞划痕和Transwell实验测定细胞迁移和侵袭,Westernblot测定上皮-间质转化(EMT)相关蛋白vimentin、Snail、Twist的表达。以无转染的Panc-1细胞为空白对照组。结果:与正常胰腺上皮细胞HPDE6-C7相比,lncRNAHOST2在各胰腺癌细胞系中的表达均明显上调(均P<0.05)。与空白对照组比较,si-HOST2组细胞增殖、迁移和侵袭能力均明显减弱,vimentin、Twist1、Snail蛋白相对表达量均明显下调(均P<0.05),而阴性对照组上述指标均无明显变化(均P>0.05)。结论:lncRNAHOST2在胰腺癌细胞中高表达,且与胰腺癌增殖、迁移和侵袭密切相关,其机制可能与调节EMT相关蛋白的表达有关。
Objective: To investigate the effect of long non-coding RNA(lncRNA) HOST2 on proliferation, migration and invasion in pancreatic cancer cells and the mechanism.Methods: The lncRNA HOST2 expressions in normal pancreatic epithelial cell line HPDE6-C7 and four different pancreatic cancer cell lines(Panc-1, AsPC-1, BxPC-3 and HPAC) were determined by qRT-PCR. Panc-1 cells were transfected with siRNA-HOST2(si-HOST2 group) or scrambled sequence(negative control group), andthen, the proliferative, migration and invasion abilities were analyzed by MTT assay, wound scratch assay and Transwell assay, and the expressions of epithelial–mesenchymal transition(EMT) associated proteins vimentin, Snail and Twist1 were measured by Western blot, respectively. The untransfected Panc-1 cells were used as blank control group.Results: The expressions of lncRNA HOST2 were significantly increased in all the studied pancreatic cancer cell lines compared with HPDE6-C7 cells(all P<0.05). Compared with blank control group, the proliferative, migration and invasion abilities were all significantly reduced, and the expression levels of vimentin, Snail and Twist1 were all significantly decreased in si-HOST2 group(all P<0.05), while all above parameters showed no significant changes in negative control group(all P>0.05).Conclusion: lncRNA HOST2 expression is up-regulated in pancreatic cancer cells, which is closely related to the proliferation, migration and invasion of pancreatic cancer, and its mechanism may be responsible for regulating the expressions of EMT-associated proteins.
Objective: To investigate the effect of long non-coding RNA(lncRNA) HOST2 on proliferation, migration and invasion in pancreatic cancer cells and the mechanism.Methods: The lncRNA HOST2 expressions in normal pancreatic epithelial cell line HPDE6-C7 and four different pancreatic cancer cell lines(Panc-1, AsPC-1, BxPC-3 and HPAC) were determined by qRT-PCR. Panc-1 cells were transfected with siRNA-HOST2(si-HOST2 group) or scrambled sequence(negative control group), andthen, the proliferative, migration and invasion abilities were analyzed by MTT assay, wound scratch assay and Transwell assay, and the expressions of epithelial–mesenchymal transition(EMT) associated proteins vimentin, Snail and Twist1 were measured by Western blot, respectively. The untransfected Panc-1 cells were used as blank control group.Results: The expressions of lncRNA HOST2 were significantly increased in all the studied pancreatic cancer cell lines compared with HPDE6-C7 cells(all P<0.05). Compared with blank control group, the proliferative, migration and invasion abilities were all significantly reduced, and the expression levels of vimentin, Snail and Twist1 were all significantly decreased in si-HOST2 group(all P<0.05), while all above parameters showed no significant changes in negative control group(all P>0.05).Conclusion: lncRNA HOST2 expression is up-regulated in pancreatic cancer cells, which is closely related to the proliferation, migration and invasion of pancreatic cancer, and its mechanism may be responsible for regulating the expressions of EMT-associated proteins.
引文
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[21]Bossart EA,Tasdemir N,Sikora MJ,et al.SNAIL is induced by tamoxifen and leads to growth inhibition in invasive lobular breast carcinoma[J].Breast Cancer Res Treat,2019.doi:10.1007/s10549-019-05161-8.[Epub ahead of print]
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[24]Meng J,Chen S,Han JX,et al.Twist1 regulates Vimentin through Cul2 circular RNA to promote EMT in hepatocellular carcinoma[J].Cancer Res,2018,78(15):4150-4162.doi:10.1158/0008-5472.CAN-17-3009.
[25]Zhang XD,Dong XQ,Xu JL,et al.Hypoxia promotes epithelialmesenchymal transition of hepatocellular carcinoma cells via inducing Twist1 expression[J].Eur Rev Med Pharmacol Sci,2017,21(13):3061-3068.
[26]崔中水,孙保存,赵楠,等.Twist1与Bcl-2协同促进肝癌细胞EMT过程中的miRNA表达谱变化[J].天津医科大学学报,2013,19(2):85-88.doi:10.3969/j.issn.1006-8147.2013.02.001.Cui ZS,Sun BC,Zhao N,et al.Variation of miRNA profile during the promotion of EMT caused by the cooperation of Twist1 and Bcl-2 in Hepatic celluler cancer[J].Journal of Tianjin Medical University,2013,19(2):85-88.doi:10.3969/j.is sn.1006-8147.2013.02.001.
[2]Bray F,Ferlay J,Soerjomataram I,et al.Global cancer statistics2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J].CA Cancer J Clin,2018,68(6):394-424.doi:10.3322/caac.21492.
[3]Rawla P,Sunkara T,Gaduputi V.Epidemiology of Pancreatic Cancer:Global Trends,Etiology and Risk Factors[J].World JOncol,2019,10(1):10-27.doi:10.14740/wjon1166.
[4]帅勇锋,占大钱,王小军,等.LncRNA SNHG15在甲状腺癌细胞中的表达及作用[J].中国普通外科杂志,2016,25(11):1590-1595.doi:10.3978/j.issn.1005-6947.2016.11.012.Shuai YF,Zhan DQ,Wang XJ,et al.Expression and action of LncRNA SNHG15 in thyroid cancer cells[J].Chinese Journal of General Surgery,2016,25(11):1590-1595.doi:10.3978/j.issn.1005-6947.2016.11.012.
[5]邢宏松,江帆,吴国俊,等.长链非编码RNA BCAR4对胰腺癌细胞增殖和凋亡的影响及机制[J].中国普通外科杂志,2018,27(3):328-334.doi:10.3978/j.issn.1005-6947.2018.03.010.Xing HS,Jiang F,Wu GJ,et al.Effect of long non-coding RNABCAR4 on proliferation and apoptosis of pancreatic carcinoma cells and the mechanism[J].Chinese Journal of General Surgery,2018,27(3):328-334.doi:10.3978/j.issn.1005-6947.2018.03.010.
[6]Wu Y,Zhang L,Wang Y,et al.Long noncoding RNA HOTAIRinvolvement in cancer[J].Tumour Biol,2014,35(10):9531-9538.doi:10.1007/s13277-014-2523-7.
[7]Zhang Y,Zhang H,Kang H,et al.Knockdown of long non-coding RNA HOST2 inhibits the proliferation of triple negative breast cancer via regulation of the let-7b/CDK6 axis[J].Int J Mol Med,2019,43(2):1049-1057.doi:10.3892/ijmm.2018.3995.
[8]Wu Y,Yuan T,Wang WW,et al.Long Noncoding RNA HOST2Promotes Epithelial-Mesenchymal Transition,Proliferation,Invasion and Migration of Hepatocellular Carcinoma Cells by Activating the JAK2-STAT3 Signaling Pathway[J].Cell Physiol Biochem,2018,51(1):301-314.doi:10.1159/000495231.
[9]Wang Q,Zhuang ZW,Cheng YM,et al.An in vitro and in vivo study of the role of long non‐coding RNA‐HOST2 in the proliferation,migration,and invasion of human glioma cells[J].IUBMB Life,2019,71(1):93-104.doi:10.1002/iub.1943.
[10]Gao Y,Meng H,Liu S,et al.LncRNA-HOST2 regulates cell biological behaviors in epithelial ovarian cancer through a mechanism involving microRNA let-7b[J].Hum Mol Genet,2014,24(3):841-852.doi:10.1093/hmg/ddu502.
[11]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.doi:10.1038/onc.2012.193.
[12]Jiao F,Hu H,Yuan C,et al.Elevated expression level of long noncoding RNA MALAT-1 facilitates cell growth,migration and invasion in pancreatic cancer[J].Oncol Rep,2014,32(6):2485-2492.doi:10.3892/or.2014.3518.
[13]Hui B,Ji H,Xu Y,et al.RREB1-induced upregulation of the lncRNA AGAP2-AS1 regulates the proliferation and migration of pancreatic cancer partly through suppressing ANKRD1 and ANGPTL4[J].Cell Death Dis,2019,10(3):207.doi:10.1038/s41419-019-1384-9.
[14]Huang R,Nie W,Yao K,et al.Depletion of the lncRNARP11-567G11.1 inhibits pancreatic cancer progression[J].Biomed Pharmacother,2019,112:108685.doi:10.1016/j.biopha.2019.108685.[Epub ahead of print]
[15]Zhang H,Feng X,Zhang M,et al.Long non-coding RNA CASC2upregulates PTEN to suppress pancreatic carcinoma cell metastasis by downregulating miR-21[J].Cancer Cell Int,2019,19:18.doi:10.1186/s12935-019-0728-y.
[16]Wang W,Li X,Meng FB,et al.Effects of the Long Non-Coding RNA HOST2 On the Proliferation,Migration,Invasion and Apoptosis of Human Osteosarcoma Cells[J].Cell Physiol Biochem,2017,43(1):320-330.doi:10.1159/000480412.
[17]An N,Cheng D.The Long Noncoding RNA HOST2 Promotes Gemcitabine Resistance in Human Pancreatic Cancer Cells[J].Pathol Oncol Res,2018.doi:10.1007/s12253-018-0486-5.[Epub ahead of print].
[18]Smith BN,Bhowmick NA.Role of EMT in Metastasis and Therapy Resistance[J].J Clin Med,2016,5(2):pii:E17.doi:10.3390/jcm5020017.
[19]Xu J,Lamouille S,Derynck R.TGF-beta-induced epithelial to mesenchymal transition[J].Cell Res,2009,19(2):156-172.doi:10.1038/cr.2009.5.
[20]Wang XL,Huang C.Difference of TGF-β/Smads signaling pathway in epithelial-mesenchymal transition of normal colonic epithelial cells induced by tumor-associated fibroblasts and colon cancer cells[J].Mol Biol Rep,2019.doi:10.1007/s11033-019-04719-5.[Epub ahead of print]
[21]Bossart EA,Tasdemir N,Sikora MJ,et al.SNAIL is induced by tamoxifen and leads to growth inhibition in invasive lobular breast carcinoma[J].Breast Cancer Res Treat,2019.doi:10.1007/s10549-019-05161-8.[Epub ahead of print]
[22]Shibue T,Weinberg RA.EMT,CSCs,and drug resistance:the mechanistic link and clinical implications[J].Nat Rev Clin Oncol,2017,14(10):611-629.doi:10.1038/nrclinonc.2017.44.
[23]Mitra A,Mishra L,Li S.EMT,CTCs and CSCs in tumor relapse and drug-resistance[J].Oncotarget,2015,6(13):10697-10711.doi:10.18632/oncotarget.4037.
[24]Meng J,Chen S,Han JX,et al.Twist1 regulates Vimentin through Cul2 circular RNA to promote EMT in hepatocellular carcinoma[J].Cancer Res,2018,78(15):4150-4162.doi:10.1158/0008-5472.CAN-17-3009.
[25]Zhang XD,Dong XQ,Xu JL,et al.Hypoxia promotes epithelialmesenchymal transition of hepatocellular carcinoma cells via inducing Twist1 expression[J].Eur Rev Med Pharmacol Sci,2017,21(13):3061-3068.
[26]崔中水,孙保存,赵楠,等.Twist1与Bcl-2协同促进肝癌细胞EMT过程中的miRNA表达谱变化[J].天津医科大学学报,2013,19(2):85-88.doi:10.3969/j.issn.1006-8147.2013.02.001.Cui ZS,Sun BC,Zhao N,et al.Variation of miRNA profile during the promotion of EMT caused by the cooperation of Twist1 and Bcl-2 in Hepatic celluler cancer[J].Journal of Tianjin Medical University,2013,19(2):85-88.doi:10.3969/j.is sn.1006-8147.2013.02.001.