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

曹娜娜; 王玲; 单保恩

doi:10.3969/j.issn.1007-3969.2014.08.001

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

更新时间：2025-12-31

- Effect of costimulatory molecule B7-H3 on the biological characteristics of esophageal cancer Eca-109 cell line
- China Oncology Vol. 24, Issue 8, Pages: 561-567(2014)
- 作者机构：
  
  河北医科大学第四医院科研中心,河北,石家庄,050011
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.1007-3969.2014.08.001
  CLC：
- Published Online：07 November 2014，
  
  Published：07 November 2014
- 稿件说明：
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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.

曹娜娜, 王玲, 单保恩. 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.
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. DOI： 10.3969/j.issn.1007-3969.2014.08.001.

曹娜娜, 王玲, 单保恩. 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. DOI： 10.3969/j.issn.1007-3969.2014.08.001.

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(P0.05)。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(P0.05)，然而其增殖能力差异无统计学意义(P0.05)。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。

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

P0.01) and untransfected group (0.128 5±0.000 2 vs 0.540 3±0.001 3

P0.01)

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

P0.05) and untransfected group (0.421 4±0.004 8 vs 0.492 1±0.014 8

P0.05). Compared with control transfected and untransfected cells

Eca-109 cell migration and invasion abilities decreased significantly (P0.05) by siRNA interference

but no significant difference was observed between their proliferative capacity (P0.05). 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.

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