Niu J T, Liu S G, Huang Y W, Li C
Department of Otorhinolaryngology Head and Neck Surgery, Second Hospital, Tianjin Medical University, Tianjin 300221, China.
Graduate School, Tianjin Medical University, Tianjin 300070, China.
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2018 Feb 7;53(2):124-130. doi: 10.3760/cma.j.issn.1673-0860.2018.02.008.
To study the roles of miR-497 and PlexinA4 in the progression of laryngeal squamous cell carcinoma. The expressions of miR-497 and PlexinA4 in fresh tumor specimens and adjacent normal mucosa tissues as well as in cell lines of laryngeal squamous cell carcinoma (LSCC) were detected with qRT-PCR and immunohistochemistry. The association of miR-497 and PlexinA4 expressions with clinicopathologic factors and their prognostic values in LSCC were evaluated PlexinA4 siRNA and pcDNA3.1 (+ )/PlexinA4 plasmid were transfected into the LSCC and measured by Transwell to evaluate their effect on the invasion of LSCC. miR-497 was low expression in LSCC, which related to pathological differentiation, while PlexinA4 mRNA was high expression in LSCC. Kaplan-Meier method showed that the prognosis of patients with high miR-497 expression was better than that of patients with low miR-497 expression (χ(2)=10.342, =0.001); . Cox regression analysis showed that miR-497 was an independent prognostic factor for LSCC. The double luciferase reporter gene showed that the variation of the fluorescence activity of wild type PlexinA4 was significantly different from that of the control group (<0.01). In Hep-2 and TU212 cell line, the number of cells with PlexinA4 siRNA passing through the compartments was 70.00±10.85 and 85.00±6.45, significantly higher than control ( values were 30.251 and 23.936, both <0.05), the number of cells with pcDNA3.1 (+ ) /PlexinA4 was 170.56±11.95 and 142.00±10.43, also significantly less than control (F values were 35.104 and 29.643, both <0.05). The expression of miR-497 in LSCC is decreased, indicating poor prognosis, which is as an independent risk factor for prognosis of LSCC. miR-497 may modulate LSCC invasion through PlexinA4.
研究miR-497和PlexinA4在喉鳞状细胞癌进展中的作用。采用qRT-PCR和免疫组织化学方法检测miR-497和PlexinA4在新鲜肿瘤标本、癌旁正常黏膜组织以及喉鳞状细胞癌(LSCC)细胞系中的表达。评估miR-497和PlexinA4表达与临床病理因素的相关性及其在LSCC中的预后价值,将PlexinA4 siRNA和pcDNA3.1(+)/PlexinA4质粒转染入LSCC细胞,通过Transwell实验检测其对LSCC侵袭能力的影响。miR-497在LSCC中呈低表达,与病理分化相关,而PlexinA4 mRNA在LSCC中呈高表达。Kaplan-Meier法显示,miR-497高表达患者的预后优于低表达患者(χ(2)=10.342,P =0.001);Cox回归分析显示,miR-497是LSCC的独立预后因素。双荧光素酶报告基因实验显示,野生型PlexinA4荧光活性变化与对照组相比差异有统计学意义(P<0.01)。在Hep-2和TU212细胞系中,转染PlexinA4 siRNA后穿膜细胞数分别为70.00±10.85和85.00±6.45,显著高于对照组(F值分别为30.251和23.936,均P<0.05);转染pcDNA3.1(+)/PlexinA4后穿膜细胞数分别为170.56±11.95和142.00±10.43,显著低于对照组(F值分别为35.104和29.643,均P<0.05)。LSCC中miR-497表达降低,提示预后不良,是LSCC预后的独立危险因素。miR-497可能通过PlexinA4调控LSCC的侵袭。