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敲低线粒体转录因子A对胃肠道间质瘤细胞生长的影响及机制研究
Effect and mechanism of knockdown of mitochondrial transcription factor A on the growth of gastrointestinal stromal tumor cells
- 局解手术学杂志 2025年34卷第2期 页码：104-109
- 作者机构：
海南医科大学第一附属医院胃肠肿瘤外科，海南 海口 570102
- 作者简介：
陈勇，E-mail：656512468@qq.com
- 基金信息：海南省卫生健康行业科研项目(21A200052)
DOI：10.11659/jjssx.03E024005
中图分类号： R735纸质出版日期：2025-02-25，
收稿日期：2024-03-01，
- 稿件说明：
移动端阅览
周江浩, 谢书海, 陈勇. 敲低线粒体转录因子A对胃肠道间质瘤细胞生长的影响及机制研究[J]. 局解手术学杂志, 2025,34(2):104-109.
ZHOU JIANG-HAO, XIE SHU-HAI, CHEN YONG. Effect and mechanism of knockdown of mitochondrial transcription factor A on the growth of gastrointestinal stromal tumor cells. [J]. Journal of regional anatomy and operative surgery, 2025, 34(2): 104-109.
周江浩, 谢书海, 陈勇. 敲低线粒体转录因子A对胃肠道间质瘤细胞生长的影响及机制研究[J]. 局解手术学杂志, 2025,34(2):104-109. DOI： 10.11659/jjssx.03E024005.
ZHOU JIANG-HAO, XIE SHU-HAI, CHEN YONG. Effect and mechanism of knockdown of mitochondrial transcription factor A on the growth of gastrointestinal stromal tumor cells. [J]. Journal of regional anatomy and operative surgery, 2025, 34(2): 104-109. DOI： 10.11659/jjssx.03E024005.
摘要
目的
2
探究敲低线粒体转录因子A（TFAM）对肿瘤相关巨噬细胞（TAMs）极化介导的胃肠道间质瘤（GIST）细胞生长的影响及机制。
方法
2
采用佛波酯和白细胞介素（IL）-4联合诱导人急性白血病单核巨噬细胞株THP-1建立体外TAMs，将慢病毒shRNA TFAM及阴性对照shRNA NC转染至TAMs，分别记为TAMs+sh-TFAM组、TAMs+sh-NC组，另取未经转染的细胞记为TAMs组。qRT-PCR检测TFAM、IL-10、精氨酸酶-1（Arg-1） mRNA表达；Western blot检测TFAM蛋白表达；流式细胞术检测CD206表达水平。将人GIST细胞株GIST-T1分为GIST-T1组、TAMs+GIST-T1组、TAMs+sh-NC+GIST-T1组、TAMs+sh-TFAM+GIST-T1组，使用含不同TAMs培养上清液的新鲜培养基处理。采用CCK-8、EdU染色以及Annexin V-FITC/PI双染法分别检测细胞增殖与凋亡水平。
结果
2
与TAMs组和TAMs+sh-NC组比较，TAMs+sh-TFAM组THP-1细胞中TFAM mRNA和蛋白表达水平均显著下调（
P
<
0.05）。与对照组比较，TAMs组THP-1细胞中CD206表达及IL-10、Arg-1 mRNA表达水平显著升高（
P
<
0.05）；与TAMs+sh-NC组比较，TAMs+sh-TFAM组THP-1细胞中CD206表达及IL-10、Arg-1 mRNA表达水平显著降低（
P
<
0.05）。与GIST-T1组比较，TAMs+GIST-T1组GIST-T1细胞增殖活性及EdU阳性细胞率均显著增加（
P
<
0.05），细胞凋亡率显著降低（
P
<
0.05）；与TAMs+sh-NC+GIST-T1组比较，TAMs+sh-TFAM+GIST-T1组GIST-T1细胞增殖活性及EdU阳性细胞率显著降低（
P
<
0.05），细胞凋亡率显著升高（
P
<
0.05）。
结论
2
敲低TFAM能够抑制TAMs向M2表型极化，从而抑制GIST细胞增殖，促进细胞凋亡，阻止GIST进展。
Abstract
Objective
2
To explore the effect and mechanism of knockdown of mitochondrial transcription factor A （TFAM） on the growth of gastrointestinal stromal tumor （GIST）cells mediated by tumor-associated macrophages （TAMs） polarization.
Methods
2
Human acute leukemia mononuclear macrophage cell line THP-1 induced by phobate and interleukin （IL）-4 was used to establish TAMs，lentivirus shRNA TFAM and negative control shRNA NC were transfected into TAMs，which were recorded as TAMs+sh-TFAM group and TAMs+sh-NC group，respectively，the untransfected cells were recorded as the TAMs group. The mRNA expression of TFAM，IL-10 and arginase-1 （Arg-1） was detected by qRT-PCR.The TFAM protein expression was detected by Western blot. The expression level of CD206 was detected by flow cytometry. The human GIST cell line GIST-T1 were divided into GIST-T1 group，TAMs+GIST-T1 group，TAMs+sh-NC+GIST-T1 group，TAMs+sh-TFAM+GIST-T1 group，and treated with fresh medium containing different TAMs culture supernatants. The cell proliferation and apoptosis levels were detected by CCK-8，EdU staining and Annexin V-FITC/PI double staining respectively.
Results
2
Compared with TAMs group and TAMs+sh-NC group，the TFAM mRNA and protein expression levels of THP-1 cells in TAMs+sh-TFAM group were significantly down-regulated （
P
<
0.05）.Compared with control group，the expression levels of CD206 and IL-10 and Arg-1 mRNA of THP-1 cells in TAMs group were significantly increased （
P
<
0.05）. Compared with TAMs+sh-NC group，the expression levels of CD206 and IL-10 and Arg-1 mRNA of THP-1 cells in TAMs+sh-TFAM group were significantly decreased （
P
<
0.05）.Compared with GIST-T1 group，the proliferative activity of GIST-T1 cells and EdU positive cells rate in TAMs+GIST-T1 group were significantly increased （
P
<
0.05），while the apoptosis rate was significantly decreased （
P
<
0.05）. Compared with TAMs+sh-NC+GIST-T1 group，the proliferative activity of GIST-T1 cells and EdU positive cells rate in TAMs+sh-TFAM+GIST-T1 group were significantly decreased （
P
<
0.05），while the apoptosis rate was significantly increased （
P
<
0.05）.
Conclusion
2
Knockdown of TFAM can inhibit the polarization of TAMs to M2 phenotype，thus inhibiting the proliferation of GIST cells，promoting cell apoptosis，and preventing the progression of GIST.
关键词
胃肠道间质瘤线粒体转录因子A肿瘤相关巨噬细胞增殖凋亡
Keywords
gastrointestinal stromal tumormitochondrial transcription factor Atumor-associated macrophagesproliferationapoptosis
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