长链非编码SSTR5-AS1对喉癌细胞侵袭迁移和PI3K/Akt通路的影响  

Effects of long non-coding SSTR5-AS1 on invasion,migration and PI3K/Akt pathway of laryngeal cancer cells

作　　者：夏兵华 杨森 王申 金建平 杨振栋 吴丹丹 XIA Binghua;YANG Sen;WANG Shen;JIN Jianping;YANG Zhendong;WU Dandan(Department of Otolaryngology, Suzhou Ninth People's Hospital, Suzhou 215200, China)

机构地区：[1]苏州市第九人民医院耳鼻喉科,江苏苏州215200

出　　处：《临床肿瘤学杂志》2020年第8期686-691,共6页Chinese Clinical Oncology

摘　　要：目的探讨长链非编码RNA SSTR5反义RNA 1(SSTR5-AS1)对喉癌细胞侵袭迁移和磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)通路的影响。方法采用实时定量PCR(qPCR)检测人鼻咽上皮细胞NP69和喉癌细胞(TU-212、TU-177和Hep-2)的SSTR5-AS1水平。构建SSTR5-AS1高表达质粒pcDNA3.1-SSTR5-AS1并采用脂质体转染Hep-2细胞(过表达组),同时设转染空pcDNA3.1的空载体组和常规培养未转染的对照组;采用MTT比色法、划痕实验和Transwell小室实验检测增殖活力、划痕愈合率和穿膜细胞数,qPCR和Western blotting检测基质金属蛋白酶(MMP)-2、MMP-9、磷酸化PI3K(p-PI3K)和磷酸化Akt(p-Akt)水平。结果喉癌细胞的SSTR5-AS1水平均低于NP69细胞(P<0.05),选取SSTR5-AS1水平最低的Hep-2细胞进行后续的过表达实验;与对照组和空载体组相比,过表达组Hep-2细胞转染pcDNA3.1-SSTR5-AS1后的SSTR5-AS1水平升高至40.691±2.287,而转染48 h的细胞活力降低(P<0.05)。过表达组Hep-2细胞的划痕愈合率和穿膜细胞数为(41.208±4.362)%和(102.095±11.173)个,均低于对照组的(79.835±5.220)%和(312.490±22.727)个及空载体组的(80.325±6.908)%和(295.617±19.634)个,差异有统计学意义(P<0.05);过表达组的MMP-2、MMP-9、p-PI3K和p-Akt水平均低于对照组和空载体组(P<0.05)。结论喉癌细胞中SSTR5-AS1低表达,且通过抑制增殖和侵袭迁移能力来发挥抑癌作用,可能与降低MMP-2、MMP-9水平和抑制PI3K/Akt通路活性有关,有望成为喉癌治疗的潜在靶点。Objective To investigate the effects of long non-coding RNA SSTR5 antisense RNA 1(SSTR5-AS1)on the invasion,migration and phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)pathway of laryngeal cancer cells.Methods The quantitative real time-polymerase chain reaction(qPCR)was used to detect SSTR5-AS1 levels in human nasopharyngeal immortalized epithelial cells NP69 and laryngeal cancer cell lines TU-212,TU-177 and Hep-2.The high expression plasmid pcDNA3.1-SSTR5-AS1 was constructed and transfected into Hep-2 cells(overexpression group)with liposome.Moreover,cells transfected with empty pcDNA3.1 were set as empty vector group and cells without transfection in conventional culture were set as control group.MTT colorimetry,scratch test and Transwell chamber test were used to detect the proliferation activity,wound healing rate and number of penetrating cells.QPCR and Western blotting were used to detect levels of matrix metalloproteinase(MMP)-2,MMP-9,phosphorylated PI3K(p-PI3K)and phosphorylated Akt(p-Akt).Results The levels of SSTR5-AS1 in laryngeal cancer cells were lower than that in NP69 cells(P<0.05).Hep-2 cells with the lowest level of SSTR5-AS1 were selected for subsequent overexpression experiment.Compared with the control group and empty vector group,the SSTR5-AS1 level in Hep-2 cells transfected with pcDNA3.1-SSTR5-AS1 in the overexpression group increased to 40.691±2.287 folds,while the cell viability decreased at 48 h after transfection(P<0.05).The wound healing rate and number of penetrating cells in Hep-2 cells of overexpression group were(41.208±4.362)%and 102.095±11.173,lower than(79.835±5.220)%and 312.490±22.727 of control group and(80.325±6.908)%and 295.617±19.634 of empty vector group(P<0.05).Levels of MMP-2,MMP-9,p-PI3K and p-Akt in the overexpression group were lower than those in the control group and empty vector group(P<0.05).Conclusion The expression of SSTR5-AS1 is low in laryngeal carcinoma cells,and it can play an anti-cancer role through inhibiting proliferation,invasion and

关 键 词：喉癌 长链非编码RNA SSTR5反义RNA 1 迁移侵袭 磷脂酰肌醇3激酶/蛋白激酶B通路 

分 类 号：R739.65[医药卫生—肿瘤]