LINC00158通过促进脾酪氨酸激酶基因的表达抑制膀胱癌细胞增殖和迁移
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摘要
目的观察LINC00158对膀胱癌细胞功能的影响,并探究其可能的分子机制。方法荧光实时定量聚合酶链反应(qPCR)检测14例膀胱癌组织及癌旁组织、膀胱癌细胞株和正常膀胱上皮细胞中LINC00158的表达量。以表达量最低的膀胱癌细胞为转染对象,转染含有LINC00158全长的质粒(实验组)或转染对照质粒(对照组);qPCR检测转染后细胞中LINC00158和脾酪氨酸激酶(SYK)mRNA的表达;免疫印迹法(Western Blot)检测SYK蛋白的表达;细胞计数试剂盒(CCK-8法)和划痕实验分别检测LINC00158对膀胱癌细胞增殖和迁移能力的影响。结果 LINC00158在膀胱癌组织中的表达明显低于癌旁组织(P <0.01),在膀胱癌细胞株中的表达明显低于正常膀胱上皮细胞(P <0.05),在T24细胞中表达量最低(P <0.01)。LINC00158在实验组细胞中的表达明显高于对照组(P <0.01)。SYK在mRNA水平和蛋白水平的表达明显升高(P <0.05)。高表达LINC00158的细胞增殖能力明显降低(P <0.05),细胞迁移能力明显降低(P <0.01)。结论 LINC00158在膀胱癌组织和细胞中低表达,高表达LINC00158可以抑制膀胱癌细胞的增殖和迁移能力,其可能的分子机制是促进SYK基因的表达。
Objective To observe the effect of LINC00158 on the function of bladder cancer cells and explore its possible molecular mechanisms. Methods Fluorescence real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression of LINC00158 in 14 cases of bladder cancer tissues,paracancerous tissues,bladder cancer cell lines and normal bladder epithelial cells. The bladder cancer cells with the lowest expression rate were transfected with full-length plasmid LINC00158(experimental group)or transfected control plasmid(control group);qPCR was used to detect the expression of LINC00158 and spleen associated tyrosine kinase(SYK)mRNA;Expression of SYK protein was detected by Western Blot;Cell counting kit(CCK-8 method)and scratch test were used to detect the effect of LINC00158 on the proliferation and migration of bladder cancer cells. Results The expression of LINC00158 in bladder cancer tissues was significantly lower than that in adjacent tissues(P < 0.01),and the expression in bladder cancer cells was significantly lower than that in normal bladder epithelial cells(P < 0.05),and the expression in T24 cells was the lowest(P < 0.01). The expression of LINC00158 in the experimental group was significantly higher than that in the control group(P < 0.01). The mRNA and protein expression levels of SYK was significantly increased(P < 0.01). The proliferation of cells with high expression of LINC00158 was significantly decreased(P < 0.05),and the cell migration ability was significantly decreased(P < 0.01). Conclusions LINC00158 is lowly expressed in bladder cancer tissues and cells. High expression of LINC00158 can inhibit the proliferation and migration of bladder cancer cells. The possible molecular mechanism is to promote the expression of SYK gene,which provides an experimental basis for gene therapy of bladder cancer.
Objective To observe the effect of LINC00158 on the function of bladder cancer cells and explore its possible molecular mechanisms. Methods Fluorescence real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression of LINC00158 in 14 cases of bladder cancer tissues,paracancerous tissues,bladder cancer cell lines and normal bladder epithelial cells. The bladder cancer cells with the lowest expression rate were transfected with full-length plasmid LINC00158(experimental group)or transfected control plasmid(control group);qPCR was used to detect the expression of LINC00158 and spleen associated tyrosine kinase(SYK)mRNA;Expression of SYK protein was detected by Western Blot;Cell counting kit(CCK-8 method)and scratch test were used to detect the effect of LINC00158 on the proliferation and migration of bladder cancer cells. Results The expression of LINC00158 in bladder cancer tissues was significantly lower than that in adjacent tissues(P < 0.01),and the expression in bladder cancer cells was significantly lower than that in normal bladder epithelial cells(P < 0.05),and the expression in T24 cells was the lowest(P < 0.01). The expression of LINC00158 in the experimental group was significantly higher than that in the control group(P < 0.01). The mRNA and protein expression levels of SYK was significantly increased(P < 0.01). The proliferation of cells with high expression of LINC00158 was significantly decreased(P < 0.05),and the cell migration ability was significantly decreased(P < 0.01). Conclusions LINC00158 is lowly expressed in bladder cancer tissues and cells. High expression of LINC00158 can inhibit the proliferation and migration of bladder cancer cells. The possible molecular mechanism is to promote the expression of SYK gene,which provides an experimental basis for gene therapy of bladder cancer.
引文
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[17]张志浩,周娟,张为民.上皮-间质转化对EGFR突变型非小细胞肺癌细胞株PC-9放射敏感性的影响[J].实用医学杂志,2017,33(10):1576-1580.
[18]薛云,李道明,张静,等.Stat5在甲状腺癌中的表达及其与上皮间质转化的关系[J].实用医学杂志,2017,33(6):905-908.2018-12-23
[2]GIELCHINSKY I,GILON M,ABU-LAIL R,et al.H19 noncoding RNA in urine cells detects urothelial carcinoma:A pilot study[J].Biomarkers,2017,22(7):661-666.
[3]DUAN W,DU L,JIANG X,et al.Identification of a serum circulating lncRNA panel for the diagnosis and recurrence prediction of bladder cancer[J].Oncotarget,2016,7(48):78850-78858.
[4]CHANDRA G S,NANDAN T Y.Potential of long non-coding RNAs in cancer patients:From biomarkers to therapeutic targets[J].Int J Cancer,2017,140(9):1955-1967.
[5]CHEN J,MIAO Z,XUE B,et al.Long non-coding RNAs in urologic malignancies:Functional roles and clinical translation[J].J Cancer,2016,7(13):1842-1855.
[6]ZHEN S,HUA L,LIU Y H,et al.Inhibition of long non-coding RNA UCA1 by CRISPR/Cas9 attenuated malignant phenotypes of bladder cancer[J].Oncotarget,2017,8(6):9634-9646.
[7]PEI Z,DU X,SONG Y,et al.Down-regulation of lncRNACASC2 promotes cell proliferation and metastasis of bladder cancer by activation of the Wnt/beta-catenin signaling pathway[J].Oncotarget,2017,8(11):18145-18153.
[8]PAN J,LI X,WU W,et al.Long non-coding RNA UCA1 promotes cisplatin/gemcitabine resistance through CREB modulating miR-196a-5p in bladder cancer cells[J].Cancer Lett,2016,382(1):64-76.
[9]ZHANG H,GUO Y,SONG Y,et al.Long noncoding RNAGAS5 inhibits malignant proliferation and chemotherapy resistance to doxorubicin in bladder transitional cell carcinoma[J].Cancer Chemother Pharmacol,2017,79(1):49-55.
[10]ZHU Y,DAI B,ZHANG H,et al.Long non-coding RNALOC572558 inhibits bladder cancer cell proliferation and tumor growth by regulating the AKT-MDM2-p53 signaling axis[J].Cancer Lett,2016,380(2):369-374.
[11]CHUANLIANG P,YUNPENG Z,YINGTAO H,et al.Syk expression in non-small-cell lung cancer and its relation with angiogenesis[J].J Cancer Res Ther,2016,12(2):663-666.
[12]PENG H,HUANG J,HU Y,et al.Associations between polymorphisms in the SYK promoter and susceptibility to sporadic colorectal cancer in a Southern Han Chinese population-a short report[J].Cell Oncol(Dordr),2015,38(2):165-172.
[13]GHOTRA V P,HE S,VAN DER HORST G,et al.SYK is a candidate kinase target for the treatment of advanced prostate cancer[J].Cancer Res,2015,75(1):230-240.
[14]LUO J,CHEN J,LI H,et al.LncRNA UCA1 promotes the invasion and EMT of bladder cancer cells by regulating the miR-143/HMGB1 pathway[J].Oncol Lett,2017,14(5):5556-5562.
[15]TAN M,ZHANG D,ZHANG E,et al.SENP2 suppresses epithelial-mesenchymal transition of bladder cancer cells through deSUMOylation of TGF-betaRI[J].Mol Carcinog,2017,56(10):2332-2341.
[16]CHEN Y,PENG Y,XU Z,et al.LncROR promotes bladder cancer cell proliferation,migration,and epithelial-mesenchymal transition[J].Cell Physiol Biochem,2017,41(6):2399-2410.
[17]张志浩,周娟,张为民.上皮-间质转化对EGFR突变型非小细胞肺癌细胞株PC-9放射敏感性的影响[J].实用医学杂志,2017,33(10):1576-1580.
[18]薛云,李道明,张静,等.Stat5在甲状腺癌中的表达及其与上皮间质转化的关系[J].实用医学杂志,2017,33(6):905-908.2018-12-23