【摘要】 目的 探讨长链非编码RNA(long non-coding RNA，lncRNA)膀胱癌相关转录因子1(bladder cancer-associated transcript 1，BLACAT1)在喉鳞状细胞癌(简称喉鳞癌)中的表达及其对喉鳞癌细胞增殖、侵袭的影响及分子机制。方法 实时荧光定量PCR(qRT-PCR)检测45例喉鳞癌组织和对应的癌旁组织中BLACAT1和miR-5590-3p的表达，分析BLACAT1表达与临床病理学参数的关系。将BLACAT1 siRNA、对照siRNA及阴性对照和miR-5590-3p模拟物转染喉鳞癌Hep-2细胞，采用qRT-PCR检测BLACAT1、miR-5590-3p和母亲DPP同源物5(果蝇)[mothers against DPP homolog 5(Drosophila)，SMAD5]的表达，CCK-8和Transwell小室检测Hep-2细胞增殖和侵袭的变化，双荧光素酶报告基因检测miR-5590-3p与BLACAT1和SMAD5之间的相互作用。结果 喉鳞癌组织中BLACAT1的表达水平(8.483±0.995)显著高于癌旁组织(1.099±0.019)，差异有统计学意义(t=5.765，P<0.001)。在有淋巴结转移的喉鳞癌组织中BLACAT1的表达水平(12.50±1.772)显著高于无淋巴结转移组织(6.268±1.997)，差异有统计学意义(t=3.319，P=0.0018)。BLACAT1的表达与淋巴结转移和组织学分级密切相关(P<0.01)。抑制BLACAT1的表达能够显著抑制Hep-2细胞的增殖和侵袭能力。miR-5590-3p在喉鳞癌组织中的表达水平显著低于癌旁组织，且miR-5590-3p表达的上调能够显著抑制Hep-2细胞的增殖和侵袭能力。此外，BLACAT1和SMAD5是miR-5590-3p的直接作用靶点，下调BLACAT1表达能促进Hep-2细胞中miR-5590-3p的表达和抑制SMAD5的表达，而miR-5590-3p模拟物能抑制Hep-2细胞中SMAD5的表达。结论 BLACAT1/miR-5590-3p/SMAD5调控轴可能在喉鳞癌细胞的增殖和侵袭中发挥重要作用，因而可能成为喉鳞癌患者潜在的治疗靶点。更多还原

【Abstract】 OBJECTIVE To explore the expression of long non-coding RNA(lncRNA) bladder cancer-associated transcript 1(BLACAT1),and its effects on cell proliferation and invasion and related molecular mechanisms in laryngeal squamous cell carcinoma(LSCC).METHODS Real-timequantitative PCR(qRT-PCR) was used to detect the expression of BLACAT1 and miR-5590-3p in 45 cases of LSCC tissues and paired normal tissues,and the association of BLACAT1 with clinicopathological features was investigated.BLACAT1 siRNA,control siRNA as well as negative control and miR-5590-3p mimic were employed to transfect into LSCC Hep-2 cell,qRT-PCR was utilized to investigate the expressions of BLACAT1,miR-5590-3p and mothers against DPP homolog 5(Drosophila) (SMAD5),and CCK-8 and Transwell chamber were used to detect the changes of cell proliferation and invasion ability of Hep-2 cell.Dual-Luciferase report experiment was used to investigate the interaction of miR-5590-3p with BLACAT1 and SMAD5.RESULTS BLACAT1 level in LSCC tissues(8.483±0.995) was significantly higher than that in normal tissues(1.099±0.019),and the difference was statistical significance(t=5.765,P<0.001).BLACAT1 level in LSCC tissues with lymph node metastasis(12.50±1.772) was significantly higher than that in normal tissues without lymph node metastasis(6.268±1.997),and the difference was statistical significance(t=3.319,P=0.0018).BLACAT1 expression was tightly associated with lymph node metastasis and histological grade(P<0.01).BLACAT1 siRNA markedly downregulated BLACAT1 level,and suppressed cell proliferation and invasion ability in Hep-2 cell.The miR-5590-3p level in LSCC tissues was dramatically lower than that in normal tissues,miR-5590-3p mimic evidently upregulated miR-5590-3p level,and its upregulation inhibited cell proliferation and invasion in Hep-2 cell.In addition,BLACAT1 and SMAD5 was the direct target of miR-5590-3p.BLACAT1 siRNA markedly upregulated miR-5590-3p level and downregulated SMAD5 level,whereas miR-5590-3pmimic decreased SMAD5 level in Hep-2 cell.CONCLUSIONBLACAT1/miR-5590-3p/SMAD5 regulatory axis may play an essential role in cell proliferation and invasion of LSCC cells,and thus may be the potential molecular therapeutic target of LSCC patients.更多还原