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miR-195 在喉鳞状细胞癌中的表达及其对 Hep-2 增殖和凋亡的影响。

Expression of miR-195 in laryngeal squamous cell carcinoma and its effect on proliferation and apoptosis of Hep-2.

机构信息

Department of Otorhinolaryngology and Maxillofacial Oncology; Tianjin Medical University Cancer Institute and Hospital, Key Laboratory of Cancer Prevention and Therapy, Tianjin Cancer Institute; National Clinical Research Center of Cancer; Tianjin, China.

出版信息

Eur Rev Med Pharmacol Sci. 2017 Jul;21(14):3232-3238.

Abstract

OBJECTIVE

To investigate the expression of miR-195 and its relationship with clinicopathological characteristics in laryngeal squamous cell carcinoma (LSCC), and to explore its effect and possible mechanism on proliferation and apoptosis of Hep-2.

PATIENTS AND METHODS

Real-time fluorescence quantitative PCR was used to detect the expression of miR-195 in laryngeal carcinoma tissues and adjacent normal tissues from 98 cases. Dual-luciferase reporter plasmid with Bcl-2 wild type and mutant type 3' untranslated region was created to verify the target of miR-195 by luciferase assay. After Hep-2 cells were transfected with miR-195/Bcl-2, miR-195, Bcl-2 siRNA and negative control by lipofectamine, the protein expression of Bcl-2 was detected by Western blot analysis. The proliferation and apoptosis of Hep-2 were detected by MTS method and flow cytometry, respectively.

RESULTS

Compared with adjacent normal tissues, the expression of miR-195 was lower in laryngeal carcinoma tissues (p < 0.01). The low expression of miR-195 was positively correlated with distant metastasis and clinical stage (p < 0.05). The average survival time of patients with low expression was shorter than those with high expression by Kaplan-Meier method (p < 0.01). Multivariate Cox analysis showed that miR-195 expression and lymph node metastases were independent prognostic factors (p < 0.05).

CONCLUSIONS

The expression of miR-195 was significantly decreased in laryngeal carcinoma tissues, which was closely related to the clinicopathological characteristics of LSCC. miR-195 may inhibit the proliferation and promote the apoptosis of Hep-2 by regulating Bcl-2 expression, which as an anti-oncogene could have the potential to be a therapeutic strategy in the treatment of LSCC.

摘要

目的

研究 miR-195 在喉鳞状细胞癌(LSCC)中的表达及其与临床病理特征的关系,并探讨其对 Hep-2 增殖和凋亡的影响及其可能的机制。

方法

采用实时荧光定量 PCR 检测 98 例喉癌组织及癌旁正常组织中 miR-195 的表达。构建带有 Bcl-2 野生型和突变型 3'UTR 的双荧光素酶报告质粒,通过荧光素酶实验验证 miR-195 的靶基因。用脂质体将 miR-195/Bcl-2、miR-195、Bcl-2 siRNA 及阴性对照转染 Hep-2 细胞后,采用 Western blot 分析检测 Bcl-2 蛋白的表达。采用 MTS 法和流式细胞术分别检测 Hep-2 细胞的增殖和凋亡。

结果

与癌旁正常组织相比,喉癌组织中 miR-195 的表达水平较低(p<0.01)。miR-195 的低表达与远处转移和临床分期呈正相关(p<0.05)。Kaplan-Meier 法显示 miR-195 低表达患者的平均生存时间短于高表达患者(p<0.01)。多因素 Cox 分析显示,miR-195 表达和淋巴结转移是独立的预后因素(p<0.05)。

结论

miR-195 在喉癌组织中表达显著下调,与 LSCC 的临床病理特征密切相关。miR-195 可能通过调节 Bcl-2 表达抑制 Hep-2 的增殖并促进其凋亡,作为一种抑癌基因,有可能成为治疗 LSCC 的一种策略。

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