共刺激分子B7-H3对食管癌细胞Eca-109生物学特性的影响

doi:10.3969/j.issn.1007-3969.2014.08.001

中国癌症杂志 ›› 2014, Vol. 24 ›› Issue (8): 561-567.doi: 10.3969/j.issn.1007-3969.2014.08.001

共刺激分子B7-H3对食管癌细胞Eca-109生物学特性的影响

曹娜娜,王玲,单保恩

河北医科大学第四医院科研中心，河北石家庄 050011

出版日期:2014-08-30 发布日期:2014-11-07
通信作者: 单保恩　E-mail:shanbaoen_1962@163.com
基金资助:
国家自然资金面上项目(No：81173611)；河北省重大医学科研课题(No：zd2013045)

Effect of costimulatory molecule B7-H3 on the biological characteristics of esophageal cancer Eca-109 cell line

CAO Na-na, WANG Ling, SHAN Bao-en

Research Center, Fourth Hospital of Hebei Medical University, Shijiazhuang Hebei 050011, China

Published:2014-08-30 Online:2014-11-07
Contact: SHAN Bao-en E-mail: shanbaoen_1962@163.com

摘要/Abstract

摘要：

背景与目的：食管肿瘤严重威胁着人类健康，认识食管癌的发生、发展机制和寻找治疗方法已经成为遏制肿瘤的研究热点。近年来，作为B7免疫球蛋白超家族的新成员，共刺激分子B7-H3在多种肿瘤中异常高表达，被认为可能是一种新的肿瘤标志物和潜在的治疗靶点。本研究旨在检测食管癌细胞株TE-1、TE-13、Eca-109细胞中B7-H3的表达，并通过靶向干扰B7-H3基因表达来研究B7-H3对食管癌细胞增殖、侵袭等生物学行为的影响。方法：运用逆转录聚合酶链反应(reverse transcription polymerasechain reaction，RT-PCR)方法检测B7-H3分子在食管癌细胞TE-1、TE-13、Eca-109中的表达。通过LipofectamineTM2000体外转染B7-H3 siRNA、control siRNA至食管癌Eca-109细胞，采用RT-PCR和蛋白质印迹法(Western blot)检测Eca-109细胞中B7-H3 mRNA和蛋白的表达。采用MTT实验、细胞划痕实验、Transwell侵袭小室实验检测B7-H3 siRNA对Eca-109细胞增殖能力、平面迁移能力及体外侵袭能力的影响。结果：共刺激分子B7-H3在食管癌细胞TE-1(0.382±0.008)、TE-13(0.399±0.008)、Eca-109(0.428±0.012)中的表达差异无统计学意义(P>0.05)。转染B7-H3 siRNA后Eca-109细胞B7-H3 mRNA和蛋白表达水平显著低于未转染组(0.128 5±0.000 2 vs 0.540 3±0.001 3，0.421 4±0.004 8 vs 0.492 1±0.014 8，P均<0.05)以及空载体转染组(0.128 5±0.000 2 vs 0.532 4±0.000 7，0.421 4±0.004 8 vs 0.500 6±0.012 9，P均<0.05)。与对照组细胞相比，转染B7-H3 siRNA后Eca-109细胞的平面迁移能力和侵袭力明显下降(P<0.05)，然而其增殖能力差异无统计学意义(P>0.05)。结论：食管癌细胞TE-1、TE-13、Eca-109均组成性表达B7-H3分子。沉默B7-H3基因表达能明显抑制Eca-109细胞的体外迁移、侵袭能力，提示B7-H3基因可能参与调节食管癌的侵袭转移能力，为免疫治疗提供潜在的治疗靶点。

关键词: 共刺激分子, B7-H3, 食管癌, Eca-109

Abstract:

Background and purpose: Esophageal cancer is a serious disease threatening human health, and it is very difficult to understand the development mechanism and find the therapeutic methods for esophageal cancer. In recent years, B7-H3, as a new member of B7 immunoregulatory superfamily, overexpressed in multiple tumor types, is considered to be a new tumor marker and potential therapeutic target. This study aimed to detect the expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13, Eca-109 and exploring the effect of B7-H3 siRNA on cell proliferation, migration and invasion in vitro in human esophageal cancer Eca-109 cell line. Methods: The expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13 and Eca-109 were detected by reverse transcription polymerase chain reaction (RT-PCR). B7-H3 siRNA and control siRNA were transfected in vitro into human esophageal cancer Eca-109 cells using LipofectamineTM 2000. The expressions of B7-H3 mRNA and protein in Eca-109 cells were analyzed by RT-PCR and Western blot. The proliferation, migration and invasion abilities of Eca-109 cells were measured by MTT assay, wound scrape assay and transwell invasion assay in vitro, respectively. Results: All tested cultured esophageal cancer cell lines constitutively expressed B7-H3 mRNA under normal conditions (TE-1 0.382±0.008, TE-13 0.399±0.008, Eca-109 0.428±0.012). After transfection, the expression of B7-H3 mRNA levels decreased in B7-H3 siRNA transfected group, compared with control siRNA transfected group (0.128 5±0.000 2 vs 0.532 4±0.000 7, P<0.01) and untransfected group (0.128 5±0.000 2 vs 0.540 3±0.001 3, P<0.01), while its protein expression levels were also significantly lower than the control transfection group (0.421 4±0.004 8 vs 0.500 6±0.012 9, P<0.05) and untransfected group (0.421 4±0.004 8 vs 0.492 1±0.014 8, P<0.05). Compared with control transfected and untransfected cells, Eca-109 cell migration and invasion abilities decreased significantly (P<0.05) by siRNA interference, but no significant difference was observed between their proliferative capacity (P>0.05). Conclusion: All tested esophageal cancer cell lines constitutively express B7-H3 mRNA. B7-H3 siRNA interference inhibits Eca-109 cell migration and invasion abilities. B7-H3 may have a critical role in regulating Eca-109 cell progression.

Key words: Costimulatory molecule, B7-H3, Esophageal cancer, Eca-109

曹娜娜,王玲,单保恩. 共刺激分子B7-H3对食管癌细胞Eca-109生物学特性的影响[J]. 中国癌症杂志, 2014, 24(8): 561-567.

CAO Na-na, WANG Ling, SHAN Bao-en. Effect of costimulatory molecule B7-H3 on the biological characteristics of esophageal cancer Eca-109 cell line[J]. China Oncology, 2014, 24(8): 561-567.

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共刺激分子B7-H3对食管癌细胞Eca-109生物学特性的影响

Effect of costimulatory molecule B7-H3 on the biological characteristics of esophageal cancer Eca-109 cell line

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