Expression of Tim4 in Glioma and its Regulatory Role in LN-18 Glioma Cells.
Expression of Tim4 in Glioma and its Regulatory Role in LN-18 Glioma Cells.
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DOI：
10.12659/msm.894963
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发表时间：
2016-01-07
期刊：
影响因子：
--
通讯作者：
Zhang Q
中科院分区：
文献类型：
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作者：
Li W;Li X;Xu S;Ma X;Zhang Q
Tim4 是一种跨膜蛋白，称为 T 细胞免疫球蛋白和含有蛋白 4 的粘蛋白结构域。我们推测 Tim4 可能与神经胶质瘤有关。本研究旨在探讨Tim4在胶质瘤中的表达水平以及Tim4对LN-18胶质瘤细胞生长和凋亡的调节作用。从神经胶质瘤患者收集肿瘤组织和邻近正常组织。分别通过 RT-PCR 和 Western blot 分析测定 Tim4 mRNA 和蛋白的表达水平，以评估其与神经胶质瘤的关联。 Tim4 在培养的人神经胶质瘤细胞 LN-18 中被 siRNA 干扰过度表达或沉默。 MTT法和流式细胞术检测LN-18细胞的生长和凋亡。通过集落形成测定评估LN-18细胞的集落形成能力。人体组织的采集经哈尔滨医科大学肿瘤医院研究伦理委员会批准，严格按照国际标准进行。所有患者均需签署知情同意书。肿瘤组织中 Tim4 mRNA 和蛋白的表达水平显着高于癌旁正常组织。靶向 Tim4 的反义 miRNA 抑制 LN-18 细胞的生长，诱导其凋亡，并降低其克隆形成能力。相反，Tim4的过度表达促进LN-18细胞的生长，抑制其凋亡，并增强其克隆形成潜力。 Tim-4的表达水平与胶质瘤密切相关。 Tim4表达减少抑制LN-18细胞的生长和集落形成能力并诱导其凋亡，而Tim4表达增加则刺激LN-18细胞的生长和克隆形成潜力并抑制其凋亡。
Tim4 is a transmembrane protein known as T cell immunoglobulin and mucin domain containing protein-4. We speculated that Tim4 might be associated with glioma. This study aimed to investigate the expression level of Tim4 in gliomas and the regulatory role of Tim4 on the growth and apoptosis of LN-18 glioma cells. Tumor tissues and adjacent normal tissues were collected from patients with glioma. The expression level of Tim4 mRNA and protein was determined by RT-PCR and Western blot analyses, respectively to evaluate their association with glioma. Tim4 was overexpressed or silenced by siRNA interference in cultured human glioma cells LN-18. The growth and apoptosis of LN-18 cells was detected by MTT assay and flow cytometry. The colony-forming ability of LN-18 cells was assessed by the colony formation assay. The collection of human tissues was approved by the Research Ethics Committee at the Harbin Medical University Cancer Hospital and performed in strict accordance with international standards. All patients were required to sign the informed consent. The expression level of Tim4 mRNA and protein in tumor tissues was significantly higher compared with adjacent normal tissues. Antisense miRNA targeting Tim4 inhibited the growth of LN-18 cells, induced their apoptosis, and reduced their clonogenic capacity. In contrast, overexpression of Tim4 promoted the growth of LN-18 cells, inhibited their apoptosis, and enhanced their clonogenic potential. The expression level of Tim-4 is closely associated with glioma. Decreased expression of Tim4 inhibited the growth and colony-forming ability of LN-18 cells and induced their apoptosis, whereas increased expression of Tim4 stimulated the growth and clonogenic potential of LN-18 cells and suppressed their apoptosis.