miRNA-135b对鼻咽癌细胞侵袭和转移的促进作用
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摘要
目的研究mi RNA-135b对鼻咽癌CNE1细胞系侵袭和转移的影响和作用机制。方法用逆转录聚合酶链反应(RT-PCR)检测mi RNA-135b在鼻咽癌CNE1细胞系中的表达,分别在鼻咽癌CNE1细胞系中过表达和下调mi RNA-135b,通过微孔滤膜培养小室及双室联合培养系统(Transwell~(TM))检测肿瘤细胞迁移和侵袭能力,采用Western blotting检测大型肿瘤抑制因子同源蛋白2(LATS2)的表达。结果 miRNA-135b在鼻咽癌CNE1细胞系中高表达,通过增强mi RNA-135b表达,鼻咽癌CNE1细胞的侵袭和转移能力有所提升;通过靶向mi RNA-135b干扰载体,下调mi RNA-135b表达,鼻咽癌CNE1细胞的侵袭和转移能力有所下降。增强miRNA-135b表达,LATS2活性和表达下降,而下调miRNA-135b表达,LATS2活性和表达上升。结论miRNA-135b过表达会提升鼻咽癌CNE1细胞的侵袭和转移能力,下调miRNA-135b表达可有效抑制鼻咽癌细胞增殖,促进细胞凋亡。miRNA-135b的表达可能通过影响LATS2活性影响鼻咽癌细胞的生长。
Objective To investigate the effect of miRNA-135 b in invasion and metastasis of nasopharyngeal carcinoma(NPC) and potential mechanisms. Methods Expression of miRNA-135 b in NPC tissues and NPC cell lines were detected by RT-PCR. The invasion and metastasis were identified by Transwell TM assay. Expression of LATS2 protein and activity of LATS2 protein were determined by western blot and LATS2 activity assay kit, respectively. Results Expression of miRNA-135 b was significantly increased in NPC cell lines. Transwell TM assay showed that over-expression of miRNA-135 b induced a dramatic improvement of invasion and metastasis in CNE1 cells, which was reversed by knockdown of miRNA-135 b. Expression and activity of LATS2 were decreased significantly by over-expression of miRNA-135 b, which was abolished by knockdown of miRNA-135. Conclusions mi RNA-135 b promotes the invasion and metastasis of nasopharyngeal carcinoma cells through mediating activity of LATS2.
Objective To investigate the effect of miRNA-135 b in invasion and metastasis of nasopharyngeal carcinoma(NPC) and potential mechanisms. Methods Expression of miRNA-135 b in NPC tissues and NPC cell lines were detected by RT-PCR. The invasion and metastasis were identified by Transwell TM assay. Expression of LATS2 protein and activity of LATS2 protein were determined by western blot and LATS2 activity assay kit, respectively. Results Expression of miRNA-135 b was significantly increased in NPC cell lines. Transwell TM assay showed that over-expression of miRNA-135 b induced a dramatic improvement of invasion and metastasis in CNE1 cells, which was reversed by knockdown of miRNA-135 b. Expression and activity of LATS2 were decreased significantly by over-expression of miRNA-135 b, which was abolished by knockdown of miRNA-135. Conclusions mi RNA-135 b promotes the invasion and metastasis of nasopharyngeal carcinoma cells through mediating activity of LATS2.
引文
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[13]JUNG H S,SEO Y R,YANG Y M,et al.Galpha 12 gep oncogene inhibits FOXO1 in hepatocellular carcinoma as a consequence of miR-135b and miR-194 dysregulation[J].Cell Signal,2014,26(7):1456-1465.
[2]张彦兵,王妍华,郭亚焕,等.miR-135b促进肝癌细胞侵袭和转移[J].细胞与分子免疫学杂志,2015,31(10):1316-1321.
[3]万仁强,傅向军,张学辉,等.miR-21在鼻咽癌中的表达及对鼻咽癌细胞株增殖及凋亡的影响[J].中国耳鼻咽喉颅底外科杂志,2015,21(5):378-381.
[4]MURAKAMI H,MIZUNO T,TANIGUCHI T,et al.LATS2 is a tumor suppressor gene of malignant mesothelioma[J].Cancer Res,2011,71(3):873-883.
[5]YANG G L,ZHANG L H,BO J J,et al.Increased expression of HMGB1is associated with poor prognosis in human bladder cancer[J].J Surg Oncol,2012,106(1):57-61.
[6]张彦兵,王妍华,郭亚焕,等.miR-135b促进肝癌细胞侵袭和转移[J].细胞与分子免疫学杂志,2015,31(10):1316-1321.
[7]TANG J F,YU Z H,LIU T,et al.Five miRNAs as novel diagnostic biomarker candidates for primary nasopharyngeal carcinoma[J].Asian Pac J Cancer Prev,2014,15(18):7575-7581.
[8]BANDRES E,CUBEDO E,AGIRRE X,et al.Identification by RealtimePCR of 13 mature microRNAs differentially expressed incolorectal cancer and non-tumoral tissues[J].Mol Cancer,2006,2(5):29-32.
[9]MELKAMU T,ZHANG X,TAN J,et al.Alteration of microRNAexpression in vinyl carbamate-induced mouse lung tumors and modulation by the chemopreventive agent indole-3-carbinol[J].Carcinogenesis,2010,31(2):252-258.
[10]ZHANG Y,HU C F,CHEN J,et al.LATS2 is de-methylated and overexpressed in nasopharyngeal carcinoma and predicts poor prognosis[J].BMC Cancer,2010(10):538.
[11]SAYED D,RANE S,LYPOWY J,et a1.MicroRNA-2l targets Sprouty 2 and promotes cellular outgrowths[J].Mol Biol Cell,2008,19(8):3272-3278.
[12]HE Y,WANG J,YUNG V Y,et al.microRNA-135b regulates apoptosis and chemoresistance in colorectal cancer by targeting large tumor suppressor kinase 2[J].Am J Cancer Res,2015,5(4):1382-1395.
[13]JUNG H S,SEO Y R,YANG Y M,et al.Galpha 12 gep oncogene inhibits FOXO1 in hepatocellular carcinoma as a consequence of miR-135b and miR-194 dysregulation[J].Cell Signal,2014,26(7):1456-1465.