STK15在头颈鳞状细胞癌组织中的表达及其对Hep-2细胞株生长的影响

doi:10.13481/j.1671-587x.20150122

摘要/Abstract

摘要：

目的: 探讨丝氨酸/苏氨酸激酶 15(STK15)在头颈鳞状细胞癌(鳞癌)组织中的表达及其与各临床病理特征的关系,阐明沉默STK15对人喉癌Hep-2细胞株生物学的影响。方法: 采用免疫组织化学方法检测头颈鳞癌组织中STK15蛋白表达;采用慢病毒沉默STK15,获得稳定转染细胞株;实验分为对照组(PLKO.1-puro-siRNA)和实验组(PLKO.1-puro-siSTK15),采用Western blotting法、RT-PCR 法、MTT法、侵袭实验和免疫荧光法检测沉默STK15对Hep-2细胞中STK15蛋白和STK15 mRNA表达水平、细胞增殖活性、穿膜细胞百分比和中心体数量的影响。结果: 免疫组织化学检测,STK15蛋白在头颈鳞癌组织中的表达率为64.96%,STK15蛋白表达与头颈鳞癌的TNM分期和预后有密切关联(P<0.05)。Western blotting法检测,实验组STK15蛋白表达水平明显降低(P<0.05)。RT-PCR检测,实验组STK15 mRNA表达水平明显降低(P<0.05)。MTT法检测,实验组细胞增殖活性明显低于对照组 (P<0.05)。侵袭实验,实验组穿膜细胞百分比明显低于对照组 (P<0.05)。免疫荧光检测,实验组中心体数目明显低于对照组 (P<0.05)。结论: STK15蛋白表达与头颈鳞癌的发展和预后密切关联,沉默STK15可降低Hep-2细胞的生长速度和迁移能力,提示STK15可作为头颈鳞癌的治疗靶点和预后标志物。

关键词: 丝氨酸/苏氨酸激酶15基因, 头颈部肿瘤, 预后

Abstract:

Objective To expore the expression of serine/threonine kinase 15 gene (STK15) in head and neck squamous cell carcinoma(HNSCC)tissue and the relationship with the clinicopathological characteristics of HNSCC, and to clarify the effect of silencing STK15 on Hep-2 cells. Methods Immunohistochemistry was used to detect the expression of STK15 protein in the HNSCC tissue.Lentiviral silencing STK15 was used in Hep-2 cell line to obtain stable transfection cell line.The Hep-2 cells were divided into control group(PLKO.1-puro-siRNA)and experimental group(PLKO.1-puro-siSTK15).Western blotting method, RT-PCR, MTT assay, cell migration assay and immunofluorescence assay were used to study the expression of STK15 protein and STK15 mRNA, cell proliferation, migration and the number of centrosome. Results Immunohistochemical staining revealed that the expression rate of STK15 protein in HNSCC tissue was 64.96%, and the expression of STK15 protein was correlated with TNM stage and prognosis(P<0.05). The results of Western blotting method showed the expression level of STK15 protein in experimental group was lower than that in control group(P<0.05).The results of RT-PCR showed the expression level of STK15 mRNA in experimental group was lower than that in control group(P<0.05).The results of proliferation assay showed the proliferation rate of the cells in experimental group was lower than that in control group(P<0.05).The results of cell migration assay showed the percentage of the cells penetrating the membrane in experimental group was lower than that that in control group(P<0.05).The results of immunofluorescence assay showed that the number of centrosome in experimental group was lower than that in control group(P<0.05). Conclusion The expression of STK15 protein is closely correlated with the development and prognosis of HNSCC and silencing STK15 can greatly reduce the proliferation and migration ability of Hep-2 cells.STK15 could represent a potentially new important therapeutic target of HNSCC, and the status of STK15 expression may also become a novel and prognostic biomarker for HNSCC patients.

Key words: serine/threonine kinase 15 gene, head and neck neoplasms, prognosis

中图分类号:

R939.9

李雅冬, 张劲松. STK15在头颈鳞状细胞癌组织中的表达及其对Hep-2细胞株生长的影响[J]. 吉林大学学报(医学版), 2015, 41(01): 114-119.

LI Yadong, ZHANG Jinsong. Expression of STK15 in head and neck squamous cell carcinoma tissue and its effect on growth of Hep-2 cells[J]. Journal of Jilin University Medicine Edition, 2015, 41(01): 114-119.

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