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1. 潍坊医学院临床医学院,山东 潍坊 261053
2. 北京大学深圳医院胸外科,广东 深圳 518036
3. 北京大学深圳医院中心实验室,广东 深圳 518036
LIU Jixian E-mail: 252110465@qq.com
收稿:2021-11-28,
纸质出版:2022-02-28
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侯清华, 钟燕凤, 刘林壮, 等. CBX3在肺腺癌中的表达、预后相关性及对癌细胞生物学行为的影响[J]. 中国癌症杂志, 2022,32(2):152-160.
Qinghua HOU, Yanfeng ZHONG, Linzhuang LIU, et al. Expression, prognostic value of CBX3 in lung adenocarcinoma and its effect on biological behavior of cancer cells[J]. China Oncology, 2022, 32(2): 152-160.
侯清华, 钟燕凤, 刘林壮, 等. CBX3在肺腺癌中的表达、预后相关性及对癌细胞生物学行为的影响[J]. 中国癌症杂志, 2022,32(2):152-160. DOI: 10.19401/j.cnki.1007-3639.2022.02.007.
Qinghua HOU, Yanfeng ZHONG, Linzhuang LIU, et al. Expression, prognostic value of CBX3 in lung adenocarcinoma and its effect on biological behavior of cancer cells[J]. China Oncology, 2022, 32(2): 152-160. DOI: 10.19401/j.cnki.1007-3639.2022.02.007.
背景与目的:
作为异染色质重要组分的染色体盒3(chromobox 3
CBX3)参与多种癌症的发生、发展过程
而
CBX
3基因在肺腺癌中的表达情况和作用尚不清楚。探讨CBX3在肺腺癌中的表达与肺腺癌患者预后的关系
并研究CBX3对A549肺腺癌细胞增殖、迁移和侵袭能力的影响。
方法:
下载癌症基因组图谱(The Cancer Genome Atlas
TCGA)中的肺腺癌转录组数据和对应的临床数据
应用R软件分析CBX3在肺腺癌中的表达差异和免疫浸润相关性
采用受试者工作特征(receiver operating characteristic
ROC)曲线和Kaplan-Meier分析CBX3在肺腺癌中的预后意义;收集北京大学深圳医院智能化生物样本库-80 ℃新鲜冻存的2020年8月
&
#x02014;2021年1月行肺癌姑息性切除术或根治性切除术的肺腺癌样本20例(已获得知情同意)
采用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction
RTFQ-PCR)检测20例冻存的肺腺癌组织及配对正常肺组织中CBX3的表达;应用小干扰RNA(siRNA)转染A549细胞以沉默CBX3的表达
并根据siRNA转染的类型将A549细胞分为沉默组(si-CBX3组)和阴性对照组(si-NC组)
通过用细胞计数试剂盒(cell counting kit-8
CCK-8)实验、克隆形成实验、划痕实验和transwell实验分别研究CBX3对A549肺腺癌细胞增殖、迁移和侵袭能力的影响。
结果:
基于TCGA肺腺癌数据的差异分析与采集的肺腺样本RTFQ-PCR实验的结果显示
CBX3在肺腺癌中呈显著的高表达
差异有统计学意义(
P
<
0.05);辅助型T细胞2的浸润与CBX3表达呈正相关
相关系数(
r
)=0.437
而肥大细胞(
r
=-0.444)、嗜酸性粒细胞(
r
=-0.380)等免疫细胞与CBX3表达呈负相关;ROC曲线下面积(area under curve
AUC)值为0.912
表明CBX3对肺腺癌具有很好的诊断价值
Kaplan-Meier分析结果显示
CBX3高表达组生存期更短
提示预后较差;沉默CBX3在A549细胞中的表达后
CCK-8实验和克隆形成实验结果表明
A549细胞的增殖能力显著降低(
P
<
0.01);划痕实验结果表明沉默后的A549细胞的迁移率为(22.68
&
#x000b1;3.44)%
较阴性对照组迁移能力明显降低(
P
<
0.05);Transwell实验结果表明A549细胞的相对侵袭率为(53.94
&
#x000b1;5.39)%
侵袭能力显著降低(
P
<
0.01)。
结论:
CBX3在肺腺癌中显著高表达并提示肺腺癌预后不良
可作为肺腺癌的预后标志物
且CBX3的高表达促进了A549细胞的增殖、迁移和侵袭能力。
Background and purpose:
Chromobox 3 (CBX3)
an important component of chromatin
is involved in the development and progression of various cancers. However
the expression and role of CBX3 in lung adenocarcinoma are still unclear. This study aimed to investigate the relationship between the expression of CBX3 in lung adenocarcinoma and prognosis of lung adenocarcinoma
and to study the effects of CBX3 on proliferation
migration and invasion of A549 lung adenoca
rcinoma cells.
Methods:
Transcriptome data of lung adenocarcinoma and corresponding clinical data were downloaded from the Cancer Genome Atlas (TCGA)
and R software was used to analyze the expression difference of CBX3 and the correlation of immune infiltration in lung adenocarcinoma. The prognostic significance of CBX3 in lung adenocarcinoma was analyzed by receiver operating characteristic (ROC) curve and Kaplan-Meier analysis. Twenty lung adenocarcinoma samples from patients (with informed consent obtained) who underwent palliative or radical lung cancer resection from August 2020 to January 2021 were collected from the intelligent Biobank of Peking University Shenzhen Hospital at -80 ℃. Real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) was used to verify the expression of CBX3 in 20 frozen lung adenocarcinoma tissue samples and paired normal lung tissues. A549 cells were transfected with small interfering RNA (siRNA) to silence the expression of CBX3
and A549 cells were divided into silent group (si-CBX3 group) and negative control group (si-NC group) according to the type of siRNA transfection. The effects of CBX3 on proliferation
migration and invasion of A549 cells were investigated by cell counting kit-8 (CCK-8) assay
clone formation assay
scratch assay and transwell assay
respectively.
Results:
The difference analysis based on TCGA lung adenocarcinoma data and RTFQ-PCR results of lung adenocarcinoma samples showed that CBX3 was significantly overexpressed in lung adenocarcinoma (
P
<
0.05). The infiltration of T helper 2 (Th2) cells was positively correlated with the expression of CBX3 [correlation coefficient (
r
)=0.437
]
while mast cells (
r
=-0.444)
eosinophils (
r
=-0.380) and other immune cells were negatively correlated with the expression of CBX3. The area under ROC curve (AUC) value was 0.912
indicating that CBX3 has a good diagnostic value for lung adenocarcinoma. Kaplan-Meier a
nalysis showed that the survival period of the group with high expression of CBX3 was shorter
suggesting a poor prognosis. After the expression of CBX3 was silenced in A549 cells
significantly decreased proliferation ability of A549 cells was detected by CCK-8 and clone formation assays (
P
<
0.01). The results of scratch assay showed that the mobility of A549 cells was (22.68
&
#x000b1;3.44)% after silencing
which was significantly lower compared with the control group (
P
<
0.05). Transwell assay results showed that the relative invasion rate of A549 cells was (53.94
&
#x000b1;5.39)%
and the invasion ability was significantly decreased (
P
<
0.01).
Conclusion:
CBX3 is significantly overexpressed in lung adenocarcinoma
which predicts poor prognosis of lung adenocarcinoma and can be used as a prognostic marker of lung adenocarcinoma
and the high expression of CBX3 promotes the proliferation
migration and invasion of A549 cells.
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